ZA200209833B - Targeted chromosomal genomic alterations in plants using modified single stranded oligonucleotides. - Google Patents

Description

TARGETED CHROMOSOMAL GENOMIC ALTERATIONS IN PLANTS

USING MODIFIED SINGLE STRANDED OLIGONUCLEOTIDES

Field Of The Invention

The technical field of the invention is oligonucleotide-directed repair or alteration of plant genetic information using novel chemically modified oligonucleotides.

Background Of The Invention

A number of methods have been developed specifically to alter the genomic information of plants. These methods generally include the use of vectors such as, for example, T-DNA, carrying nucleic acid sequences encoding partial or complete portions of a particular protein which is expressed in a cell or tissue to effect the alteration. The expression of the particular protein then results in the desired phenotype. See, for example, United States Patent 4,459,355 which describes a method for transforming plants with a DNA vector and United States Patent 5,188,642 which describes cloning or expression vectors containing a transgenic DNA sequence which when expressed in plants confers resistance to the herbicide glyphosate. The use of such fransgene-containing vectors adds one or more exogenous copies of a gene in a usually random fashion at one or more integration sites of the plant's genome at some variable frequency. The introduced gene may be foreign or may be derived from the host plant. Any gene which was originally present in the genome, which may be, for example, a normal allelic variant, mutated, defective, and/or functional copy of the introduced gene, is retained in the genome of the host plant.

These methods of gene alteration are problematic in that complications which can compromise the vigor, productivity, yield, etc. of the plant may result. One such problem is that insertion of exogenous nucleic acid at random location(s) in the genome can have deleterious effects. The random . nature of this insertion and/or the use of exogenous promoters can also cause the timing, location or strength of expression of the introduced transgene to be inappropriate or unpredictable. Another problem with such systems includes the addition of unnecessary and unwanted genetic material to the genome of - the recipient, including, for example, T-DNA ends or other vector remnants, exogenous control sequences required to allow production of the transgene protein, which control sequences may be a exogenous or native to the host plant and/or the transgene, and reporter genes or resistance markers.

Such remnants and added sequences may have presently unrecognized consequences, for example, involving genetic rearrangements of the recipient genomes. In addition, concerns have been raised with consumption, especially by humans, of plants containing such exogenous genetic material. } 5 More recently, simpler systems involving poly- or oligo- nucleotides have been described for use in the alteration of genomic DNA. These chimeric RNA-DNA oligonucleotides, requiring contiguous RNA and DNA bases in a double-stranded molecule folded by complementarity into a double hairpin conformation, have been shown to effect single basepair or frameshift alterations, for example, for ‘mutation or repair of plant, animal or fungal genomes. See, for example, WO 99/07865 and U.S. Patent 5,565,350. In the chimeric RNA-DNA oligonucleotide, an uninterrupted stretch of DNA bases within the molecule is required for sequence alteration of the targeted genome while the obligate RNA residues are involved in complex stability. Due to the length, backbone composition, and structural configuration of these chimeric RNA-DNA molecules, they are expensive to synthesize and difficult to purify. Moreover, if the RNA-containing strand of the chimeric RNA-DNA oligonucleotide is designed so as to direct gene alteration, a series of mutagenic reactions resulting in nonspecific base alteration can result. Such a result reduces the utility of such a molecule in methods designed for targeted gene alteration.

Alternatively, other oligo- or poly- nucleotides have been used which require a triplex forming, usually polypurine or polypyrimidine, structural domain which binds to a DNA helical duplex through Hoogsteen interactions between the major groove of the DNA duplex and the oligonucleotide.

Such oligonucleotides may have an additional DNA reactive moiety, such as psoralen, covalently linked to the oligonucleotide. These reactive moieties function as effective intercalation agents, stabilize the formation of a triplex and can be mutagenic. Such agents may be required in order to stabilize the triplex forming domain of the oligonucleotide with the DNA double helix if the Hoogsteen interactions from the oligonucleotide/target base composition are insufficient. See, e.g., U.S. Patent 5,422,251. The utility of these oligonucleotides for directing targeted gene alteration is compromised by a high frequency of nonspecific base changes. : In more recent work, the domain for altering a genome is linked or tethered to the triplex . forming domain of the bi-functional oligonucleotide, adding an additional linking or tethering functional domain to the oligonucleotide. See, e.g., Culver et al., Nature Biotechnology 17: 989-93 (1999). Such i 30 chimeric or triplex forming molecules have distinct structural requirements for each of the different domains of the complete poly- or oligo-nucleotide in order to effect the desired genomic alteration in either episomal or chromosomal targets.

Other genes, e.g. CFTR, have been targeted by homologous recombination using duplex fragments having several hundred basepairs. See, e.g., Kunzelmann et al., Gene Ther. 3:859-867 . (1996). Similar efforts to target genes by homologous recombination in plants using large fragments of

DNA had some success. See Kempin et al., Nature 389:802-803 (1997). However, the efficiency and reproducibility of the published homologous recombination approach in plants has severely limited the widespread use of this method.

Earlier experiments to mutagenize an antibiotic resistance indicator gene by homologous recombination used an unmodified DNA oligonucleotide rather than larger fragments of DNA, wherein the oligonucleotide had no functional domains other than a region of complementary sequence to the target.

See Campbell et al., New Biologist 1: 223-227 (1989). These experiments required large concentrations of the oligonucleotide, exhibited a very low frequency of episomal modification of a targeted exogenous plasmid gene not normally found in the cell and have not been reproduced. However, as shown in examples herein, we have observed that an unmodified DNA oligonucleotide can convert a base at low frequency which is detectable using the assay systems described herein.

Oligonucleotides designed for use in the targeted alteration of genetic information are significantly different from oligonucleotides designed for antisense approaches. For example, antisense oligonucleotides are perfectly complementary to and bind an mRNA strand in order to modify expression of a targeted mRNA and are used at high concentration. As a consequence, they are unable to produce a gene conversion event by either mutagenesis or repair of a defect in the chromosomal DNA of a host genome. Furthermore, the backbone chemical composition used in most oligonucleotides designed for use in antisense approaches renders them inactive as substrates for homologous pairing or mismatch repair enzymes and the high concentrations of oligonucleotide required for antisense applications can be toxic with some types of nucleotide modifications. In addition, antisense oligonucleotides must be complementary to the mRNA and therefore, may not be complementary to the other DNA strand or to genomic sequences that span the junction between intron sequence and exon sequence.

Artificial chromosomes can be useful for the screening purposes identified herein. These molecules are man-made linear or circular DNA molecules constructed from essential cis-acting DNA sequence elements that are responsible for the proper replication and partitioning of natural chromosomes (Murray et al., 1983). The essential elements are: (1) Autonomous Replication Sequences (ARS), (2) Centromeres, and (3) Telomeres.

Yeast artificial chromosomes (YACs) allow large segments of genomic DNA to be cloned and modified (Burke et al., Science 236:806; Peterson et al., Trends Genet. 13:61 (1997); Choi, etal, Nat.

Genet, 4:117-223 (1993), Davies, et al., Biotechnology 11:911-914 (1993), Matsuura, et al., Hum. Mol.

Genet., 5:451-459 (1996), Peterson et al., Proc. Natl. Acad. Sci., 93:6605-6609 (1996); and Sched, et al., . Cell, 86:71-82 (1996)). Other vectors also have been developed for the cloning of large segments of genomic DNA, including cosmids, and bacteriophage P1 (Sternberg et al., Proc. Natl. Acad. Sci. U.S.A, : 5 87:103-107 (1990)). YACs have certain advantages over these alternative large capacity cloning vectors (Burke et al., Science, 236:806-812 (1987). The maximum insert size is 35-30 kb for cosmids, and 100 kb for bacteriophage P1, both of which are much smaller than the maximal insert size for a YAC.

An alternative to YACs are cloning systems based on the E. coli fertility factor that have been developed to construct large genomic DNA insert libraries. They are bacterial artificial chromosomes (BACs) and P-1 derived artificial chromosomes (PACs) (Mejia et al., Genome Res. 7:179-186 (1997); Shizuya et al., Proc. Natl. Acad. Sci. 89:8794-8797 (1992); loannou et al., Nat. Genet, 6:84-89 (1994); Hosoda et al., Nucleic Acids Res. 18:3863 (1990)). BACs are based on the E. coli fertility plasmid (F factor); and PACs are based on the bacteriophage P1. These vectors propagate at a very low copy number (1-2 per cell) enabling genomic inserts up to 300 kb in size to be stably maintained in recombination deficient hosts. The PACs and BACs are circular DNA molecules that are readily isolated from the host genomic background by classical alkaline lysis (Birnboim et al., Nucleic Acids Res. 7:1513-1523 (1979)). In addition, BACs have been developed for transformation of plants with high- molecular weight DNA using the T-DNA system (Hamilton, Gene 24:107-116 (1997); Frary & Hamilton,

Transgenic Res. 10: 121-132 (2001)).

A need exists for simple, inexpensive oligonucleotides capable of producing targeted alteration of genetic material such as those described herein as well as methods to identify optimal oligonucleotides that accurately and efficiently alter target DNA.

Summary Of The Invention

Novel, modified single-stranded nucleic acid molecules that direct gene alteration in plants are identified and the efficiency of alteration is analyzed both in vitro using a cell-free extract assay and in vivo using a yeast system and a plant system. The alteration in an oligonucleotide of the invention may comprise an insertion, deletion, substitution, as well as any combination of these. Site specific alteration of DNA is not only useful for studying function of proteins in vivo, but it is also useful for creating plants with desired phenotypes, including, for example, environmental stress tolerance, improved nutritional value, herbicide resistance, disease resistance, modified oil production, modified starch production, and altered floral morphology including selective sterility. As described herein,

oligonucleotides of the invention target directed specific gene alterations in genomic double-stranded

DNAin cells. The target genomic DNA can be nuclear chromosomal DNA as well as plastid or . mitochondrial chromosomal DNA. The target DNA can also be a transgene present in the plant cell, : including; for example, a previously introduced T-DNA. For screening purposes, the target plant DNA can ‘5 also be extrachromosomal DNA present in plant or non-plant cells in various forms including, e.g., mammalian artificial chromosomes (MACs), PACs from P-1 vectors, yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), plant artificial chromosomes (PLAC), as well as episomal DNA, including episornal DNA from an exogenous source such as a plasmid or recombinant vector. Many of these artificial chromosome constructs containing plant DNA can be obtained from a variety of sources, including, e.g., the Arabidopsis Biological Resource Center (ABRC) at the Ohio State University, and the

Rice Genome Research Program at the MAFF DNA bank in Ibaraki, Japan. The target DNA may be transcriptionally silent or active. In a preferred embodiment, the target DNA to be altered is the non- transcribed strand of a genomic DNA duplex. In a more preferred embodiment, the target DNA to be altered is the non-transcribed strand of a transcribed gene of a genomic DNA duplex.

The low efficiency of targeted gene alteration obtained using unmodified DNA - oligonucleotides is believed to be largely the result of degradation by nucleases present in the reaction mixture or the target cell. Although different modifications are known to have different effects on the nuclease resistance of oligonucleotides or stability of duplexes formed by such oligonucleotides (see, e.g., Koshkin et al., J. Am. Chem. Soc., 120:13252-3), we have found that itis not possible to predict which of any particular known modification would be most useful for any given alteration event, including for the construction of gene alteration oligonucleotides, because of the interaction of different as yet unidentified proteins during the gene alteration event. Herein, a variety of nucleic acid analogs have been developed that increase the nuclease resistance of oligonucleotides that contain them, including, e.g., nucleotides containing phosphorothioate linkages or 2'-0-methyl analogs. We recently discovered that single-stranded DNA oligonucleotides modified to contain 2'-0O-methyl RNA nucleotides or phosphorothioate linkages can enable specific alteration of genetic information at a higher level than either unmodified single-stranded DNA or a chimeric RNA/DNA molecule. See, for example, copending applications United States application no. 60/208,538, United States application no. 60/244,989, United

States application no. 09/818,875 international application no. PCT/US01/09761 and Gamper et al.,

Nucleic Acids Research 28: 4332-4339 (2000), the disclosures of which are incorporated herein in their entirety by reference. We also found that additional nucleic acid analogs which increase the nuclease resistance of oligonucleotides that contain them, including, e.g., “locked nucleic acids” or "LNAS", xylo- * US 2003/0051270 which incorporates the disclosures of 60/208538 and 60/244989 * WO 01/73002

Amended Sheet — 2004-08-20

LNAs and L-ribo-LNAs; see, for example, Wengel & Nielsen, WO 99/14226; Wengel, WO 00/56748;

Wengel, WO 00/66604; and Jakobsen & Koshkin, WO 01/25478 also allow specific targeted alteration of genetic information.

The assay allows for determining the optimum length of the oligonucleotide, optimum . 5 sequence of the oligonucleotide, optimum position of the mismatched base or bases, optimum chemical modification or modifications, optimum strand targeted for identifying and selecting the most efficient oligonucleotide for a particular gene alteration event by comparing to a control oligonucleotide. Control oligonucleotides may include a chimeric RNA-DNA double hairpin oligonucleotide directing the same gene alteration event, an oligonucleotide that matches its target completely, an oligonucleotide in which all linkages are phosphorothiolated, an oligonucleotide fully substituted with 2'-O-methyl analogs or an RNA oligonucleotide. Such control oligonucleotides either fail to direct a targeted alteration or do so at a lower efficiency as compared to the oligonucleotides of the invention. The assay further allows for determining the optimum position of a gene alteration event within an oligonucleotide, optimum concentration of the selected oligonucleotide for maximum alteration efficiency by systematically testing a range of concentrations, as well as optimization of either the source of cell extract by testing different plants or strains, or testing cells derived from different plants or strains, or plant cell lines. Using a series of single-stranded oligonucleotides, comprising all RNA or DNA residues and various mixtures of the two, + several new structures are identified as viable molecules in nucleotide conversion to direct or repair a genomic mutagenic event. When extracts from mammalian, plant and fungal cells are used and are analyzed using a genetic readout assay in bacteria, single-stranded oligonucleotides having one of several modifications are found to be more active than a control RNA-DNA double hairpin chimera structure when evaluated using an in vitro gene repair assay. Similar results are also observed in vivo using yeast, mammalian and plant cells. Molecules containing various lengths of modified bases were found to possess greater activity than unmodified single-stranded DNA molecules.

Detailed Description Of The Invention

The present invention provides oligonucleotides having chemically modified, nuclease . resistant residues, preferably at or near the termini of the oligonucleotides, and methods for their identification and use in targeted alteration of plant genetic material, including gene mutation, targeted gene repair and gene knockout. The oligonucleotides are preferably used for mismatch repair or alteration by changing at least one nucleic acid base, or for frameshift repair or alteration by addition or deletion of at least one nucleic acid base. The oligonucleotides of the invention direct any such alteration,

including gene correction, gene repair or gene mutation and can be used, for example, to introduce a polymorphism or haplotype or to eliminate (‘knockout’) a particufar protein activity. For example, gene } alterations that knockout a particular protein activity can be obtained using oligonucleotides designed to convert a codon in the coding region of the protein to a stop codon, thus prematurely terminating translation of the protein. Oligonucleotides that introduce stop codons in the open-reading-frame of the protein are one embodiment of the invention. Generally, oligonucleotides that introduce stop codons early in the open-reading-frame of the protein are preferred. If the open-reading-frame contains more than one methionine, oligonucleotides that introduce stop codons after the second methionine are preferred. Additionally, if the gene exhibits alternative splice sites, oligonucleotides that introduce stop codons in exons after the alternative splice site are preferred. The following table provides examples of codons that can be converted to stop codons by altering a single oligonucleotide. A skilled artisan could + readily identify other codons that can be converted to stop codons by altering one, two or three of the base pairs in a given codon. Similarly, a skilled artisan could readily identify codons that can be converted to stop codons by a frameshift mutations that inserts or deletes one or two base pairs in the open-reading-frame. [tis also understood that more than one stop codon can be generated in a single open-reading-frame and that these stop codons can be adjacent in the sequence or separated by intervening codons. Where more than one stop codon is introduced info a single open-reading-frame, such alterations can be generated by a single or multiple oligonucleotides and can be generated simultaneously or by sequential mutagenesis of the target nucleic acid. :

GGA (glycine), AGA (arginine), CGA (arginine), TTA (leucine), TGA

TCA (serine), TGT (cysteine), TGG (tryptophan), TGC (cysteine)

AAG (lysine), GAG (glutamate), CAG (glutamine), TTG (leucine),

TCG (serine), TGG (tryptophan), TAT (cysteine), TAC (tyrosine) i TCA (serine), TAT (cysteine), TAC (tyrosine) *The amino acid encoded by the original codon is shown in parentheses and the base targeted for : alteration to convert the codon to the corresponding stop codon is underlined and in bold

The oligonucleotides of the invention are designed as substrates for homologous pairing and repair enzymes and as such have a unique backbone composition that differs from chimeric RNA- . DNA double hairpin oligonucleotides, antisense oligonucleotides, and/or other poly- or oligo-nucleotides used for altering genomic DNA, such as triplex forming oligonucleotides. The single-stranded oligo- nucleotides described herein are inexpensive to synthesize and easy to purify. In side-by-side comparisons, an optimized single-stranded oligonucleotide comprising modified residues as described herein is significantly more efficient than a chimeric RNA-DNA double hairpin oligonucleotide in directing a base substitution or frameshift mutation in a cell-free extract assay.

We have discovered that single-stranded oligonucleotides having a DNA domain surrounding the targeted base, with the domain preferably central to the poly- or oligo-nucleotide, and having at least one modified end, preferably at the 3' terminal region, are able to alter a target genetic sequence and with an efficiency that is higher than chimeric RNA-DNA double hairpin oligonucleotides disclosed in US Patent 5,565,350. Preferred oligonucleotides of the invention have at least two modified bases on at least one of the termini, preferably the 3' terminus of the oligonucleotide. Oligonucleotides of the invention can efficiently be used to introduce targeted alterations in a genetic sequence of DNA in the presence of human, animal, plant, fungal (including yeast) proteins and in cells of different types including, for example, plant cells, fungal cells including S. cerevisiae, Ustillago maydis, Candida albicans, and mammalian cells. Particularly preferred are cells and cell extracts derived from plants including, for example, experimental model plants such as Chlamydomonas reinhardtii, Physcomitrella patens, and

Arabidopsis thaliana in addition to crop plants such as cauliflower (Brassica oleracea), artichoke (Cynara scolymus), fruits such as apples (Malus, e.g. domesticus), mangoes (Mangifera, e.g. indica), banana (Musa, e.g. acuminata), berries (such as currant, Ribes, e.g. rubrum), kiwifruit (Actinidia, e.g. chinensis), grapes (Vitis, e.g. vinifera), bell peppers (Capsicum, e.g. annuum), cherries (such as the sweet cherry, . Prunus, e.g. avium), cucumber (Cucumis, e.g. sativus), melons (Cucumis, e.g. melo), nuts (such as walnut, Juglans, e.q. regia; peanut, Arachis hypogeae), orange (Citrus, e.g. maxima), peach (Prunus, e.g. persica), pear (Pyra, e.g. communis), plum (Prunus, e.g. domestica), strawberry (Fragana, e.g. moschata or vesca), tomato (Lycopersicon, e.g. esculentum); leaves and forage, such as alfalfa (Medicago, €.9. ’ "sativa or truncatula), cabbage (e.g. Brassica oleracea), endive (Cichoreum, e.g. endivia), leek (Allium, e.g. porrum), lettuce (Lactuca, e.g. sativa), spinach (Spinacia, e.g. oleraceae), tobacco (Nicotiana, e.g. tabacum); roots, such as arrowroot (Maranfa, e.g. arundinacea), beet (Befa, e.g. vulgaris), carrot a. (Daucus, e.g. carota), cassava (Manihot, e.g. esculenta), turnip (Brassica, e.g. rapa), radish (Raphanus, e.g. sativus), yam (Dioscorea, e.g. esculenta), sweet potato (lpomoea batatas); seeds, including oilseeds,

such as beans (Phaseolus, e.g. vulgaris), pea (Pisum, e.g. sativum), soybean (Glycine, e.g. max), cowpea (Vigna unguiculata), mothbean (Vigna aconitifolia), wheat (Triticum, e.g. aestivum), sorghum (Sorghum e.g. bicolor), barley (Hordeum, e.g. vulgare), corn (Zea, e.g. mays), rice (Oryza, e.g. Sativa), rapeseed (Brassica napus), millet (Panicum sp.), sunflower (Helianthus annuus), oats (Avena sativa), chickpea (Cicer, e.g. arietinum); tubers, such as kohirabi (Brassica, e.g. oleraceae), potato (Solanum, e.g. tuberosum) and the like; fiber and wood plants, such as flax (Linum e.g. usitatissimum), cotton (Gossypium e.g. hirsutum), pine (Pinus sp.), oak (Quercus sp.), eucalyptus (Eucalyptus sp.), and the like and ornamental plants such as turfgrass (Lolium, e.g. rigidum), petunia (Petunia, e.g. x hybrida), hyacinth (Hyacinthus orientalis), carnation (Dianthus e.g. caryophyllus), delphinium (Delphinium, e.q. ajacis), Job's tears (Coix lacryma-jobi), snapdragon (Antirrhinum majus), poppy (Papaver, e.g. nudicaule), lilac (Syringa, e.g. vulgaris), hydrangea (Hydrangea e.g. macrophylla), roses (including Gallicas, Albas,

Damasks, Damask Perpetuals, Centifolias, Chinas, Teas and Hybrid Teas) and ornamental goldenrods (e.g. Solidago spp.). Such plant cells can then be used to regenerate whole plants according to methods described herein or any method known in the art. The DNA domain of the oligonucleotides is preferably fully complementary to one strand of the gene target, except for the mismatch base or bases responsible for the gene alteration event(s). On either side of the preferably central DNA domain, the contiguous bases may be either RNA bases or, preferably, are primarily DNA bases. The central DNA domain is generally atleast 8 nucleotides in length. The base(s) targeted for alteration in the most preferred embodiments are at least about 8, 9 or 10 bases from one end of the oligonucleotide.

According to certain embodiments, one or both of the termini of the oligonucleotides of the present invention comprise phosphorothioate modifications, LNA backbone (including LNA derivatives and analogs) modifications, or 2'-O-methyl base analogs, or any combination of these modifications.

Oligonucleotides comprising 2'-O-methyl or LNA analogs are a mixed DNA/RNA polymer. The oligonucleotides of the invention are, however, single-stranded and are not designed to form a stable internal duplex structure within the oligonucleotide. The efficiency of gene alteration is surprisingly increased with oligonucleotides having internal complementary sequence comprising phosphorothioate modified bases as compared to 2-0-methyl modifications. This result indicates that specific chemical . interactions are involved between the converting oligonucleotide and the proteins involved in the conversion. The effect of other such chemical interactions to produce nuclease resistant termini using - 30 modifications other than LNA (including LNA derivatives or analogs), phosphorothioate linkages, or 2'-O- methyl analog incorporation into an oligonucleotide can not yet be predicted because the proteins involved in the alteration process and their particular chemical interaction with the oligonucleotide substituents are not yet known and cannot be predicted. : In the examples, oligonucleotides of defined sequence are provided for alteration of genes in particular plants. Provided the teachings of the instant application, one of skill in the art could readily design oligonucleotides to introduce analogous alterations in homologous genes from any plant.

Furthermore, in the tables of these examples, the oligonucleotides of the invention are not limited to the particular sequences disclosed. The oligonucleotides of the invention include extensions of the appropriate sequence of the longer 120 base oligonucleotides which can be added base by base to the smallest disclosed oligonucleotides of 17 bases. Thus the oligonucleotides of the invention include for each correcting change, oligonucleotides of length 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120 with further single-nucleotide additions up to the longest sequence disclosed. in some embodiments, longer nucleic acids of up to 240 bases which comprise the sequences disclosed herein may be used. Moreover, the aligonucleotides of the invention do not require a symmetrical extension on either side of the central DNA domain. Similarly, the oligonucleotides of the invention as disclosed in the various tables for alteration of particular plant genes contain phosphorothioate linkages, 2-O-methy!f analog or LNA (including LNA derivatives and analogs) or any combination of these modifications just as the assay oligonucleotides do.

The present invention, however, is not limited to oligonucleotides that contain any particular nuclease resistant modification. Oligonucleotides of the invention may be altered with any combination of additional LNAs (including LNA derivatives and analogs), phosphorothioate linkages or 2"

O-methyl analogs to maximize conversion efficiency. For oligonucleotides of the invention that are longer than about 17 to about 25 bases in length, internal as well as terminal region segments of the backbone may be altered. Alternatively, simple fold-back structures at each end of a oligonucleotide or appended end groups may be used in addition to a modified backbone for conferring additional nuclease resistance. : The different oligonucleotides of the present invention preferably contain more than one of the aforementioned backbone modifications at each end. In some embodiments, the backbone modifications are adjacent to one another. However, the optimal number and placement of backbone modifications for any individual oligonucleotide will vary with the length of the oligonucleotide and the particular type of backbone modification(s) that are used. If constructs of identical sequence having phosphorothioate linkages are compared, 2, 3, 4, 5, or 6 phosphorothioate linkages at each end are preferred. If constructs of identical sequence having 2'-O-methyl base analogs are compared, 1, 2, 3 or 4 analogs are preferred. The optimal number and type of backbone modifications for any particular oligo- nucleotide useful for altering target DNA may be determined empirically by comparing the alteration : 5 efficiency of the oligonucleotide comprising any combination of the modifications to a control molecule of comparable sequence using any of the assays described herein. The optimal position(s) for oligonucleotide modifications for a maximally efficient altering oligonucleotide can be determined by testing the various modifications as compared to control molecule of comparable sequence in one of the assays disclosed herein. In such assays, a control molecule includes, e.g., a completely 2-O-methyl substituted molecule, a completely complementary oligonucleotide, or a chimeric RNA-DNA double hairpin.

Increasing the number of phosphorothioate linkages, LNAs or 2'-O-methyl bases beyond the preferred number generally decreases the gene repair activity of a 25 nucleotide long oligonucleotide.

Based on analysis of the concentration of oligonucleotide present in the extract after different time periods of incubation, itis believed that the terminal modifications impart nuclease resistance to the oligonucleo- tide thereby allowing it to survive within the cellular environment. However, this may not be the only possible mechanism by which such modifications confer greater efficiency of conversion. For example, as disclosed herein, certain modifications to oligonucleotides confer a greater improvement to the efficiency of conversion than other modifications. oo Efficiency of conversion is defined herein as the percentage of recovered substrate molecules that have undergone a conversion event. Depending on the nature of the target genetic material, e.g. the genome of a cell, efficiency could be represented as the proportion of cells or clones containing an extrachromosomal element that exhibit a particular phenotype. Alternatively, representative samples of the target genetic material can be sequenced to determine the percentage that have acquired the desire change. The oligonucleotides of the invention in different embodiments can alter DNA two, three, four, five, six, seven, eight, nine, ten, twelve, fifteen, twenty, thirty, and fifty or more fold more than control oligonucleotides. Such control oligonucleotides are oligonucleotides with fully phosphorothiolated ‘ linkages, oligonucleotides that are fully substituted with 2'-O-methyl analogs, a perfectly matched oligonucleotide that is fully complementary to a target sequence or a chimeric DNA-RNA double hairpin

C30 oligonucleotide such as disclosed in US Patent 5,565,350.

In addition, for a given oligonucleotide length, additional modifications interfere with the ability of the oligonucleotide to act in concert with the cellular recombination or repair enzyme machinery which is necessary and required to mediate a targeted substitution, addition or deletion event in DNA. For example, fully phosphorothiolated or fully 2-O-methylated molecules are inefficient in targeted gene . alteration.

The oligonucleotides of the invention as optimized for the purpose of targeted alteration : 5 of genetic material, including gene knockout or repair, are different in structure from antisense oligo- nucleotides that may possess a similar mixed chemical composition backbone. The oligonucleotides of the invention differ from such antisense oligonucleotides in chemical composition, structure, sequence, and in their ability to alter genomic DNA. Significantly, antisense oligonucleotides fail to direct targeted gene alteration. The oligonucleotides of the invention may target either strand of DNA and can include any component of the genome including, for example, intron and exon sequences. The preferred embodiment of the invention is a modified oligonucleotide that binds to the non-transcribed strand of a genomic DNA duplex. In other words, the preferred oligonucleotides of the invention target the sense strand of the DNA, i.e. the oligonucleotides of the invention are complementary to the non-transcribed strand of the target duplex DNA. The sequence of the non-transcribed strand of a DNA duplex is found in the mRNA produced from that duplex, given that mRNA uses uracil-containing nucleotides in place of thymine-containing nucleotides. :

Moreover, the initial observation that single-stranded oligonucleotides comprising these modifications and lacking any particular triplex forming domain have reproducibly enhanced gene alteration activity in a variety of assay systems as compared to a chimeric RNA-DNA double-stranded hairpin control or single-stranded oligonucleotides comprising other backbone modifications was surprising. The single-stranded molecules of the invention totally lack the complementary RNA binding structure that stabilizes a normal chimeric double-stranded hairpin of the type disclosed in

U.S. Patent 5,565,350 yet is more effective in producing targeted base conversion as compared to such a chimeric RNA-DNA double-stranded hairpin. In addition, the molecules of the invention lack any particular triplex forming domain involved in Hoogsteen interactions with the DNA double helix and required by other known oligonucleotides in other oligonucleotide-dependant gene conversion systems.

Although the lack of these functional domains was expected to decrease the efficiency of an alteration in ‘ a sequence, just the opposite occurs: the efficiency of sequence alteration using the modified oligonucleotides of the invention is higher than the efficiency of sequence alteration using a chimeric

C30 RNA-DNA hairpin targeting the same sequence alteration. Moreover, the efficiency of sequence : alteration or gene conversion directed by an unmodified oligonucleotide is many times lower as compared to a control chimeric RNA-DNA molecule or the modified oligonucleotides of the invention targeting the same sequence alteration. Similarly, molecules containing at least 3 2'-O-methyl base analogs are about four to five fold less efficient as compared to an oligonucleotide having the same number of : phosphorothioate linkages.

The oligonucleotides of the present invention for alteration of a single base are about 17 to about 121 nucleotides in length, preferably about 17 to about 74 nucleotides in length. Most preferably, however, the oligonucleotides of the present invention are at least about 25 bases in length, unless there are self-dimerization structures within the oligonucleotide. If the oligonucleotide has such an unfavorable structure, lengths longer than 35 bases are preferred. Oligonucleotides with modified ends both shorter and longer than certain of the exemplified, modified oligonucleotides herein function as gene repair or gene knockout agents and are within the scope of the present invention.

Once an oligomer is chosen, it can be tested for its tendency to self-dimerize, since self- dimerization may result in reduced efficiency of alteration of genetic information. Checking for self- dimerization tendency can be accomplished manually or, preferably, using a software program. One such program is Oligo Analyzer 2.0, available through Integrated DNA Technologies (Coralville, 1A 52241) (http://www. idtdna.com); this program is available for use on the world wide web at http://www. idtdna.com/program/oligoanalyzer/ oligoanalyzer.asp.

For each oligonucleotide sequence input into the program, Oligo Analyzer 2.0 reports possible self- dimerized duplex forms, which are usually only partially duplexed, along with the free energy change associated with such self-dimerization. Delta G-values that are negative and large in magnitude, indicating strong self-dimerization potential, are automatically flagged by the software as "bad". Another software program that analyzes oligomers for pair dimer formation is Primer Select from DNASTAR, Inc., 1228 S. Park St., Madison, WI 53715, Phone: (608) 258-7420 (http://www.dnastar.com/products/PrimerSelect. html).

If the sequence is subject to significant self-dimerization, the addition of further sequence flanking the "repair" nucleotide can improve gene correction frequency.

Generally, the oligonucleotides of the present invention are identical in sequence to one strand of the target DNA, which can be either strand of the target DNA, with the exception of one or more targeted bases positioned within the DNA domain of the oligonucleotide, and preferably toward the middle between the modified terminal regions. Preferably, the difference in sequence of the oligonucleotide as compared to the targeted genomic DNA is located at about the middle of the oligonucleotide sequence.” in a preferred embodiment, the oligonucleotides of the invention are complementary to the non-transcribed strand of a duplex. In other words, the preferred oligonucleotides target the sense strand of the DNA, i.e. the oligonucleotides of the invention are preferably complementary to the strand of the target DNA the sequence of which is found in the mRNA.

The oligonucleotides of the invention can include more than a single base change. In an oligonucleotide that is about a 70-mer, with at least one modified residue incorporated on the ends, as disclosed herein, multiple bases can be simultaneously targeted for change. The target bases may be up to 27 nucleotides apart and may not be changed together in all resultant plasmids in all cases. Thereis a frequency distribution such that the closer the target bases are to each other in the central DNA domain. within the oligonucleotides of the invention, the higher the frequency of change in a given cell. Target bases only two nucleotides apart are changed together in every case that has been analyzed. The farther apart the two target bases are, the less frequent the simultaneous change. Thus, oligonucleotides of the invention may be used to repair or alter multiple bases rather than just one single base. For example, in a 74-mer oligonucleotide having a central base targeted for change, a base change event up to about 27 nucleotides away can also be effected. The positions of the altering bases within the oligonucleotide can be optimized using any one of the assays described herein. Preferably, the altering bases are at least about 8 nucleotides from one end of the oligonucleotide.

The oligonucleotides of the present invention can be introduced into cells by any suitable means. According to certain preferred embodiments, the modified oligonucleotides may be used alone.

Suitable means, however, include the use of polycations, cationic lipids, liposomes, polyethylenimine (PEI), electroporation, biolistics, microinjection and other methods known in the art to facilitate cellular uptake. For plant cells, biolistic or particle bombardment methods are typically used. According to certain preferred embodiments of the present invention, isolated plant cells are treated in culture according to the methods of the invention, to mutate or repair a target gene. ‘Alternatively, plant target DNA may be modified in vitro or in another cell type, including for example, yeast or bacterial cells and then introduced into a plant cell as, for example, a T-DNA. Plant cells thus modified may be used to regenerate the whole organism as, for example, in a plant having a desired targeted genomic change. In other instances, targeted genomic alteration, including repair or mutagenesis, may take place in vivo following direct ’ administration of the modified, single-stranded oligonucleotides of the invention to a subject.

The single-stranded, modified oligonucleotides of the present invention have numerous

C30 applications as gene repair, gene modification, or gene knockout agents. Such oligonucleotides may be advantageously used, for example, to introduce or correct multiple point mutations. Each mutation leads to the addition, deletion or substitution of at least one base pair. The methods of the present invention offer distinct advantages over other methods of altering the genetic makeup of an organism, in that only the individually targeted bases are altered. No additional foreign DNA sequences are added to the genetic complement of the organism. Such agents may, for example, be used to develop plants with improved fraits by rationally changing the sequence of selected genes in isolated cells and using these modified cells to regenerate whole plants having the altered gene. See, e.g., U.S. Patent 6,046,380 and

U.S. Patent 5,905,185 incorporated herein by reference. Such plants produced using the compositions of the invention lack additional undesirable selectable markers or other foreign DNA sequences. Targeted base pair substitution or frameshift mutations introduced by an oligonucleotide in the presence of a cell- + free extract also provides a way to modify the sequence of extrachromosomal elements, including, for example, plasmids, cosmids and artificial chromosomes. The oligonucleotides of the invention also simplify the production of piants having particular modified or inactivated genes. Altered plant model systems such as those produced using the methods and oligonucleotides of the invention are invaluable in determining the function of a gene and in evaluating drugs. The oligonucleotides and methods of the present invention may also be used to infroduce molecular markers, including, for example, SNPs,

RFLPs, AFLPs and CAPs.

The purified oligonucleotide compositions may be formulated in accordance with routine procedures depending on the target. For example, purified oligonucleotide can be used directly in a standard reaction mixture to introduce alterations into targeted DNA in vitro or where cells are the target as a composition adapted for bathing cells in culture or for microinjection into cells in culture. The purified oligonucleotide compositions may also be provided on coated microbeads for biolistic delivery into plant cells. Where necessary, the composition may also include a solubilizing agent. Generally, the ingredients 'will be supplied either separately or mixed together in single-use form, for example, as a dry, lyophilized powder or water-free concentrate. In general, dosage required for efficient targeted gene alteration will range from about 0.001 to 50,000 g/kg target tissue, preferably between 1 to 250 pg/kg, and most preferably at a concentration of between 30 and 60 micromolar.

For cell administration, direct injection into the nucleus, biolistic bombardment, electroporation, liposome transfer and calcium phosphate precipitation may be used. In yeast, lithium ) acetate or spheroplast transformation may also be used. In a preferred method, the administration is performed with a liposomal transfer compound, e.g., DOTAP (Boehringer-Mannheim) or an equivalent such as lipofectin. The amount of the oligonucleotide used is about 500 nanograms in 3 micrograms of

DOTAP per 100,000 cells. For electroporation, between 20 and 2000 nanograms of oligonucleotide per million cells to be electroporated is an appropriate range of dosages which can be increased to improve

28-12-2001 01939797 efficiency of genetic alteration upon review of the appropriate sequence according to the methods described herein. For biolistic delivery, microbeads are generally coated with resuspended : oligonucleotides, which range of oligonucleotide to microbead concentration can be similarly adjusted to improve efficiency as determined using one of the assay methods described herein, starting with about 0.05 to 1 microgram of oligonucleotide to 25 microgram of 1.0 micrometer gold beads or similar microcarrier.

Another aspect of the invention is a kit comprising at least one oligonucleotide of the invention. The kit may comprise an additional reagent or article of manufacture. The additional reagent or article of manufacture may comprise a delivery mechanism, cell extract, a cell, or a plasmid, such as one of those disclosed in the Figures herein, for use in an assay of the invention. Altematively, the @® invention includes a kit comprising an isogenic set of cells in which each cell in the kit comprises a different altered amino acid for a target protein encoded by a targeted altered gene within the cell produced according to the methods of the invention.

Brief Description Of The Drawings

Figure 1. Flow diagram for the generation of modified single-stranded oligonucleotides.

The upper strands of chimeric oligonucleotides | and Il are separated into pathways resulting in the generation of single-stranded oligonucleotides that contain (A) 2'-O-methyl RNA nucleotides or (B) phosphorothioate linkages. Fold changes in repair activity for correction of kan® in the HUH7 cell-free extract are presented in parenthesis. HUH? cells are described in Nakabayashi et al., Cancer Research 42: 3858-3863 (1982). Each single-stranded oligonucleotide is 25 bases in length and contains a G ® residue mismatched to the complementary sequence of the kan® gene. The numbers 3, 6, 8, 10, 12 and 12.5 respectively indicate how many phosphorothioate linkages (S) or 2'-O-methyl RNA nucleotides (R) are at each end of the molecule. Hence oligo 12S/25G contains an all phosphorothioate backbone, displayed as a dotted line. Smooth lines indicate DNA residues, wavy lines indicate 2'-O-methyl RNA -’ residues and the carat indicates the mismatched base site (G). Figure 1(C) provides a schematic plasmid indicating the sequence of the kan chimeric double-stranded hairpin oligonucleotide (left; SEQ ID NO: . 2673) and the sequence the tet chimeric double-stranded hairpin oligonucleotide used in other experiments (right; SEQ ID NO: 2674). Figure 1(D) provides a flow chart of a kan experiment in which a chimeric double-stranded hairpin oligonucleotide (SEQ 1D NO: 2673) is used. In Figure 1(D), the Kan. mutant sequence corresponds to SEQ ID NO; 2675 and SEQ ID NO: 2676; the Kan converted sequence corresponds to SEQ ID NO: 2677 and SEQ ID NO: 2678; the mutant sequence in the sequence trace

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= N = - -— EEE EP ———— — 28-12-2001 01939797 -16A- corresponds to SEQ ID NO: 2679 and the converted sequences in the sequence trace correspond to

SEQ ID NO: 2680.

Figure 2. Genetic readout system for correction of a point mutation in plasmid pK*m4021. A mutant kanamycin gene harbored in plasmid pK*m4021 is the target for correction by oligonucleotides.

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28-12-2001 01939797

The mutant Gis converted to a C by the action of the oligo. Corrected plasmids confer resistance to kanamycin in E.coli (DH10B) after electroporation leading to the genetic readout and colony counts. The wild type sequence corresponds to SEQ ID NO: 2681.

Figure 3: Target plasmid and sequence correction of a frameshift mutation by chimeric : and single-stranded oligonucleotides. (A) Plasmid pT*A208 contains a single base deletion mutation at position 208 rendering it unable to confer tet resistance. The target sequence presented below indicates the insertion of a T directed by the oligonucleotides to re-establish the resistant phenotype. (B) DNA sequence confirming base insertion directed by Tet 35/256; the yellow highlight indicates the position of frameshift repair. The wild type sequence corresponds to SEQ ID NO: 2682, the mutant sequence corresponds to SEQ ID NO: 2683 and the converted sequence corresponds to SEQ ID NO: 2684. The . [ control sequence in the sequence trace corresponds to SEQ ID NO: 2685 and the 3S/25A sequence in the sequence trace corresponds to SEQ ID NO: 2686.

Figure 4. DNA sequences of representative kan’ colonies. Confirmation of sequence alteration directed by the indicated molecule is presented along with a table outlining codon distribution.

Note that 10S/25G and 12S/25G elicit both mixed and unfaithful gene repair. The number of clones sequenced is listed in parentheses next to the designation for the single-stranded oligonucleotide. A plus (+) symbol indicates the codon identified while a figure after the (+) symbol indicates the number of colonies with a particular sequence. TAC/TAG indicates a mixed peak. Representative DNA sequences are presented below the table with yellow highlighting altered residues. The sequences in the sequence traces have been assigned numbers as follows: 3S/25G, 65/25G and 8S/25G correspond to SEQ 1D NO: 2687, 108/25G corresponds to SEQ ID NO: 2688, 25S/25G on the lower left corresponds to SEQ ID NO:

J 2689 and 25S/25G on the lower right corresponds to SEQ ID NO: 2690.

Figure 5. Gene correction in HeLa cells. Representative oligonucleotides of the invention are co-transfected with the pCMVneo()FIAsH plasmid (shown in Figure 9) into HeLa cells.

Ligand is diffused into cells after co-transfection of plasmid and oligonucleotides. Green fluorescence indicates gene correction of the mutation in the antibiotic resistance gene. Correction of the mutation results in the expression of a fusion protein that carries a marker ligand binding site and when the fusion protein binds the ligand, a green fluorescence is emitted. The ligand is produced by Aurora Biosciences and can readily diffuse into cells enabling a measurement of corrected protein function; the protein must . bind the ligand directly to induce fluorescence. Hence cells bearing the corrected plasmid gene appear green while “uncorrected” cells remain colorless.

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28-12-2001 01939797 -17A -

Figure 6. Z-series imaging of corrected cells. Serial cross-sections of the Hela cell represented in Figure 5 are produced by Zeiss 510 LSM confocal microscope revealing that the fusion protein is contained within the cell. ’ Figure 7. Hygromycin-eGFP target plasmids. (A) Plasmid pAURHYG(ins)GFP contains a single base insertion mutation between nucleotides 136 and 137, at codon 46, of the Hygromycin B coding sequence (cds) which is transcribed from the constitutive ADH1 promoter. The target sequence presented below indicates the deletion of an A and the substitution of a C for a T directed by the oligonucleotides to re-establish the resistant phenotype. In Figure 7A, the sequence of the normal allele corresponds to SEQ ID NO: 2691, the sequence of the target/existing mutation corresponds to SEQ ID

NO: 2692 and the sequence of the desired alteration corresponds to SEQ ID NO: 2693. (B) Plasmid

C pAURHYG(rep)GFP contains a _

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28-12-2001 01939797 base substitution mutation introducing a G at nucleotide 137, at codon 48, of the Hygromycin B coding sequence (cds). The target sequence presented below the diagram indicates the amino acid } conservative replacement of G with C, restoring gene function. In Figure 7B, the sequence of the normal allele correspond to SEQ ID NO: 2691, the sequence of the target/existing mutation corresponds to SEQ . ID NO: 2694 and the sequence of the desired alteration comresponds to SEQ ID NO: 2693.

Figure 8. Oligonucleotides for correction of hygromycin resistance gene. The sequence of the oligonucleotides used in experiments to assay correction of a hygromycin resistance gene are shown. DNA residues are shown in capital letters, RNA residues are shown in lowercase and nucleotides with a phosphorothioate backbone are capitalized and underlined. In Figure 8, the sequence of

HygE3T/25 corresponds to SEQ ID NO: 2695, the sequence of HygE3T/74 corresponds to SEQ ID NO:

C 269, the sequence of HygE3T/74a corresponds to SEQ ID NO: 2697, the sequence of HygGG/Rev corresponds to SEQ ID NO: 2698 and the sequence of Kan70T corresponds to SEQ ID NO: 2699.

Figure 9. pAURNeo(-)FIAsH plasmid. This figure describes the plasmid structure, target sequence, oligonucleotides, and the basis for detection of the gene alteration event by fluorescence. In

Figure 9, the sequence of the Neo/kan target mutant corresponds to SEQ ID NO: 2675 and SEQ ID NO: 2676, the converted sequence corresponds to SEQ ID NO: 2677 and SEQ ID NO: 2678 and the FIAsH peptide sequence corresponds to SEQ ID NO: 2700.

Figure 10. pYESHyg(x)eGFP plasmid. This plasmid is a construct similar to the

PAURHYg(x)eGFP construct shown in Figure 7, except the promoter is the inducible GAL1 promoter.

This promoter is inducible with galactose, leaky in the presence of raffinose, and repressed in the presence of dextrose. ® Figure 11. pBI-HygeGFP plasmid. This plasmid is a construct based on the plasmids pBI101, pBl 101.2, pBI101.3 or pBI 121 available from Clontech in which HygeGFP replaces the beta- glucuronidase gene of the Clontech plasmids. The different Clontech plasmids vary by a reading frame shift relative to the polylinker, or the presence of the Cauliflower mosaic virus promoter.

The following examples are provided by way of illustration only, and are not intended to limit the scope of the invention disclosed herein.

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28-12-2001 01939797 -18A -

EXAMPLE 1

Assay Method For Base Alteration

And Preferred Oligonucleotide Selection ) In this example, single-stranded and double-hairpin oligonucleotides with chimeric backbones (see Figure 1 for structures (A and B) and sequences (C and D) of assay oligonucleotides) are used to correct a point mutation in the kanamycin gene of pk’m4021 (Figure 2) or the tetracycline gene of pT°A208 (Figure 3). All kan oligonucleotides share the same 25 base sequence surrounding the target base identified for change, just as all tet oligonucleotides do. The sequence is given in Figures 1C and

Figure 1D. Each plasmid contains a functional ampicillin gene. Kanamycin gene function is restored ® when a G at position 4021 is converted to a C (via a substitution mutation); tetracycline gene function is restored when a deletion at position 208 is replaced by a C (via frameshift mutation). A separate plasmid, pAURNeo(-)FIAsH (Figure 9), bearing the kan® gene is used in the cell culture experiments. This plasmid was constructed by inserting a synthetic expression cassette containing a neomycin phosphotransferasea

AMENDED SHEET

28-12-2001 01939797 (kanamycin resistance) gene and an extended reading frame that encodes a receptor for the FIASH ligand into the pAUR123 shuttle vector (Panvera Corp., Madison, WI). The resulting construct replicates in S. cerevisiae at low copy number, confers resistance to aureobasidinA and constitutively expresses either the Neo+/FlAsH fusion product (after alteration) or the truncated Neo-/FiAsH product (before . alteration) from the ADH1 promoter. By extending the reading frame of this gene to code for a unique peptide sequence capable of binding a small ligand to form a fluorescent complex, restoration of expression by correction of the stop codon can be detected in real time using confocal microscopy.

Additional constructs can be made to test additional gene alteration events or for specific use in different expression systems. For example, altemative comparable plant plasmids or integration vectors such as, e.g. those based on T-DNA, can be constructed for stable expression in plant cells according to ® the disclosures herein. Such constructs would use a plant specific promoter such as, e.g., cauliflower mosaic virus 35S promoter, to replace the promoters directing expression of the neo, hyg or aureobasidinA resistance gene disclosed herein, including for example, in Figures 7B, 9 and 10 herein.

Moreover, the green fluorescent protein (GFP) sequence used herein may be modified to increase expression in plant cells such as Arabidopsis and the other plants disclosed herein as described in

Haseloff et al., Proc. Natl. Acad. Sci. 94(6): 2122-7 (1997), Rouwendal et al. Plant Mol. Biol. 33(6): 989-99 (1997) and Hu et al. FEBS Lett. 369(2-3): 331-4 (1995). Codon usage for optimal expression of GFP in plants results from increasing the frequency of codons with a C or a G in the third position from 32 to about 60%. Specific constructs are disclosed and can be used as follows with such plant specific alterations.

We also construct three mammalian expression vectors, pHyg(rep)eGFP, pHyg{A)eGFP, ® pHyg(ins)eGFP, that contain a substitution mutation at nucleotide 137 of the hygromycin-B coding sequence. (rep) indicates a T137=3G replacement, (A) represents a deletion of the G137 and (ins) represents an A insertion between nucleotides 136 and 137. All point mutations create a nonsense termination codon at residue 46. We use pHYGeGFP plasmid (Invitrogen, CA) DNA as a template to introduce the mutations info the hygromycin-eGFP fusion gene by a two step site-directed mutagenesis

PCR protocol. First, we generate overlapping 5' and a 3' amplicons surrounding the mutation site by . PCR for each of the point mutation sites. A 215 bp 5' amplicon for the (rep), (A) or (ins) was generated by polymerization from oligonucleotide primer HygEGFPf (5'-AATACGACTCACTATAGG-3" SEQ ID NO: : 2701) to primer Hygrepr (5'GACCTATGCACGCCCTCC-3'; SEQ ID NO: 2702), HygAr (5-

GACTATCCACGCCCTCC-3'; SEQ ID NO: 2703), or Hyginsr (5-GACATTATCCACGCCCTCC-34 SEQ ID

NO: 2704), respectively. We generate a 300bp 3' amplicon for the (rep), (A) or (ins) by polymerization from oligonucleotide primers Hygrepf (5-CTGGGATAGGTCCTGCGG-3'; SEQ ID NO: 2705), HygAf

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28-12-2001 01939797 (5'-CGTGGATAGTCCTGCGG-3'; SEQ ID NO: 2706), Hyginsf (5-CGTGGATAATGTCCTGCGG-3'; SEQ

ID NO: 2707), respectively to primer HygEGFPr (5-AAATCACGCCATGTAGTG-3'"; SEQ ID NO: 2708).

We mix 20 ng of each of the resultant 5' and 3' overlapping amplicon mutation sets and use the mixture as a template to amplify a 523 bp fragment of the Hygromycin gene spanning the Kpnl and Rr! . restriction endonuclease sites. We use the Expand PCR system (Roche) to generate all amplicons with cycles of denaturing at 94°C for 10 seconds, annealing at 55°C for 20 seconds and elongation at 68°C for 1 minute. We digest 10 pg of vector pHYGeGFP and 5 jg of the resulting fragments for each mutation with Kpnl and Rsril (NEB) and gel purify the fragment for enzymatic ligation. We ligate each mutated insert into pHYGeGFP vector at 3:1 molar ratio using T4 DNA ligase (Roche). We screen clones by restriction digest, confirm the mutation by Sanger dideoxy chain termination sequencing and purify the

C plasmid using a Qiagen maxiprep kit.

Oligonucleotide synthesis and cells. Chimeric oligonucleotides and single-stranded oligonucleotides (including those with the indicated modifications) are synthesized using available phosphoramidites on controlled pore glass supports. After deprotection and detachment from the solid support, each oligonucleotide is gel-purified using, for example, procedures such as those described in

Gamper et al., Biochem. 39, 5808-5816 (2000) and the concentrations determined spectrophotometrically (33 or 40 pg/ml per A, unit of single-stranded or hairpin oligomer). HUH? cells are grown in DMEM, 10% FBS, 2mM glutamine, 0.5% pen/strep. The E.coli strain, DH10B, is obtained from Life Technologies (Gaithersburg, MD); DH10B cells contain a mutation in the RECA gene (recA).

Cell-free extracts. Although this portion of this example is directed to mammalian systems, similar extracts from plants can be prepared as disclosed elsewhere in this application and used ® as disclosed in this example. We prepare cell-free extracts from HUH? cells or other mammalian cells, as follows. We employ this protocol with essentially any mammalian cell including, for example, H1299 cells (human epithelial carcinoma, non-small cell lung cancer), C1271 (immortal murine mammary epithelial cells), MEF (mouse embryonic fibroblasts), HEC-1-A (human uterine carcinoma), HCT15 (human colon cancer), HCT116 (human colon carcinoma), LoVo (human colon adenocarcinoma), and Hela (human cervical carcinoma). We harvest approximately 2x10° cells. We then wash the cells immediately in cold hypotonic buffer (20 mM HEPES, pH7.5; 5 mM KCI; 1.5 mM MgCl; 1 mM DTT) with 250 mM sucrose.

We then resuspend the cells in cold hypotonic buffer without sucrose and after 15 minutes we lyse the : cells with 25 strokes of a Dounce homogenizer using a tight fitting pestle. We incubate the lysed cells for 60 minutes on ice and centrifuge the sample for 15 minutes at 12000xg. The cytoplasmic fraction is enriched with nuclear proteins due to the extended co-incubation of the fractions following cell breakage.

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--

We then immediately aliquote and freeze the supernatant at-80°C. We determine the protein concentration in the extract by the Bradford assay. } : We also perform these experiments with cell-free extracts obtained from fungal cells, including, for example, S. cerevisiae (yeast), Ustilago maydis, and Candida albicans. For example, we grow yeast cells into log phase in 2L YPD medium for 3 days at 30°C. We then centrifuge the cultures at 5000xg, resuspend the pellets in a 10% sucrose, 50 mM Tris, ImM EDTA lysis solution and freeze them on dry ice. After thawing, we add KCl, spermidine and lyticase to final concentrations of 0.25 mM, 5 mM and 0.1 mg/ml, respectively. We incubate the suspension on ice for 60 minutes, add PMSF and Triton

X100 to final concentrations of 0.1 mM and 0.1% and continue to incubate on ice for 20 minutes. We centrifuge the lysate at 3000xg for 10 minutes to remove larger debris. We then remove the supernatant and clarify it by centrifuging at 30000xg for 15 minutes. We then add glycerol to the clarified extract to a concentration of 10% (v/v) and freeze aliquots at -80°C. We determine the protein concentration of the extract by the Bradford assay.

Reaction mixtures of 50 yl are used, consisting of 10-30 pg protein of cell-free extract, which can be optionally substituted with purified proteins or enriched fractions, about 1.5 pig chimeric double-hairpin oligonucleotide or 0.55 ug single-stranded molecule (35/25G or 6S/25G, see Figure 1), and 1 pg of plasmid DNA (see Figures 2 and 3) in a reaction buffer of 20 mM Tris, pH 7.4, 15 mM MgCl,, 0.4 mM DTT, and 1.0 mM ATP. Reactions are initiated with extract and incubated at 30°C for 45 min.

The reaction is stopped by placing the tubes on ice and then immediately deproteinized by two phenol/chioroform (1:1) extractions. Samples are then ethanol precipitated. The nucleic acid is pelleted at 15,000 r.p.m. at 4°C for 30 min., is washed with 70% ethanol, resuspended in 50 pl H,0, and is stored at-20°C. 5 pl of plasmid from the resuspension (~100 ng) was transfected in 20 ul of DH10B cells by electroporation (400 V, 300 uF, 4 kQ) in a Cell-Porator apparatus (Life Technologies). After electro- poration, cells are transferred to a 14 mi Falcon snap-cap tube with 2 ml SOC and shaken at 37°C for 1 h.

Enhancement of final kan colony counts is achieved by then adding 3 ml SOC with 10 pg/ml kanamycin and the cell suspension is shaken for a further 2 h at 37°C. Cells are then spun down at 3750 x g and the pellet is resuspended in 500 ui SOC. 200 wi is added undiluted to each of two kanamycin (50 pg/ml) agar plates and 200 pi of a 10° dilution is added to an ampicillin (100 g/m) plate. After overnight 37°C incubation, bacterial colonies are counted using an Accucount 1000 (Biologics). Gene conversion effectiveness is measured as the ratio of the average of the kan colonies on both plates per amp colonies multiplied by 10° to correct for the amp dilution.

The following procedure can also be used. 5 pl of resuspended reaction mixtures (total volume 50 pl) are used to transform 20 yl aliquots of electro-competent DH10B bacteria using a : Cell-Porator apparatus (Life Technologies). The mixtures are allowed to recover in 1 mi SOC at 37°C for 1 hour at which time 50 ug/ml kanamycin or 12 ug/ml tetracycline is added for an additional 3 hours. : 5 Prior fo plating, the bacteria are pelleted and resuspended in 200 u1 of SOC. 100 pl aliquots are plated onto kan or tet agar plates and 100 pl of a 10 dilution of the cultures are concurrently plated on agar plates containing 100 g/ml of ampicillin. Plating is performed in triplicate using sterile Pyrex beads.

Colony counts are determined by an Accu-count 1000 plate reader (Biologics). Each plate contains 200- 500 ampicillin resistant colonies or 0-500 tetracycline or kanamycin resistant colonies. Resistant colonies are selected for plasmid extraction and DNA sequencing using an ABI Prism kit on an ABI 310 capillary sequencer (PE Biosystems).

Chimeric single-stranded oligonucleotides. In Figure 1 the upper strands of chimeric oligonucleotides | and Il are separated into pathways resulting in the generation of single-stranded oligo- nucleotides that contain (Figure 1A) 2'-O-methyl RNA nucleotides or (Figure 1B) phosphorothioate linkages. Fold changes in repair activity for correction of kan® in the HUH? cell-free extract are presented in parenthesis. Each single-stranded oligonucleotide is 25 bases in length and contains a G residue mismatched to the complementary sequence of the kan® gene.

Molecules bearing 3, 6, 8, 10 and 12 phosphorothioate linkages in the terminal regions at each end of a backbone with a total of 24 linkages (25 bases) are tested in the kan® system. Alternatively, molecules bearing 2, 4, 5, 7, 9 and 11 inthe terminal regions at each end are tested. The results of one such experiment, presented in Table 1 and Figure 1B, illustrate an enhancement of correction activity directed by some of these modified structures. In this illustrative example, the most efficient molecules contained 3 or 6 phosphorothioate linkages at each end of the 25-mer; the activities are approximately equal (molecules 1X and X with results of 3.09 and 3.7 respectively). A reduction in alteration activity may be observed as the number of modified linkages in the molecule is further increased. Interestingly, a single-strand molecule containing 24 phosphorothioate linkages is minimally active suggesting that this backbone modification when used throughout the molecule supports only a low level of targeted gene repair or alteration. Such a non-altering, completely modified molecule can provide a baseline control for determining efficiency of correction for a specific oligonucleotide molecule of known sequence in defining the optimum oligonucleotide for a particular alteration event.

The efficiency of gene repair directed by phosphorothioate-modified, single-stranded molecules, in a length dependent fashion, led us to examine the length of the RNA modification used in the original chimera as it relates to correction. Construct lil represents the "RNA-containing" strand of chimera | and, as shown in Table 1 and Figure 2A, it promotes inefficient gene repair. But, as shown in the same figure, reducing the RNA residues on each end from 10 to 3 increases the frequency of repair.

At equal levels of modification, however, 25-mers with 2'-0O-methyl ribonucleotides were less effective > 5 gene repair agents than the same oligomers with phosphorothioate linkages. These results reinforce the fact that an RNA containing oligonucleotide is not as effective in promoting gene repair or alteration as a modified DNA oligonucleotide.

Repair of the kanamycin mutation requires a G=#C exchange. To confirm that the specific desired correction alteration was obtained, colonies selected at random from multiple experiments are processed and the isolated plasmid DNA is sequenced. As seen in Figure 4, colonies generated through the action of the single-stranded molecules 3S/25G (IX), 6S/25G (X) and 8S/25G (XI) respectively contained plasmid molecules harboring the targeted base correction. While a few colonies appeared on plates derived from reaction mixtures containing 25-mers with 10 or 12 thioate linkages on both ends, the sequences of the plasmid molecules from these colonies contain nonspecific base changes. In these illustrative examples, the second base of the codon is changed (see Figure 3). These results show that modified single-strands can direct gene repair, but that efficiency and specificity are reduced when the 25-mers contain 10 or more phosphorothioate linkages at each end.

In Figure 1, the numbers 3, 6, 8, 10, 12 and 12.5 respectively indicate how many phosphorothioate linkages (S) or 2-0-methyt RNA nucleotides (R) are at each end of the examplified molecule although other molecules with 2, 4, 5, 7, 9 and 11 modifications at each end can also be tested.

Hence oligo 125/25G represents a 25-mer oligonucleotide which contains 12 phosphorothioate linkages on each side of the central G target mismatch base producing a fully phosphorothioate linked backbone, displayed as a dotted line. The dots are merely representative of a linkage in the figure and do not depict the actual number of linkages of the oligonucleotide. Smooth lines indicate DNA residues, wavy lines indicate 2'-O-methyl RNA residues and the carat indicates the mismatched base site (G).

Correction of a mutant kanamycin gene in cultured mammalian cefls. Although this portion of this example is directed to cultured mammalian cells, comparable methods may be used using ’ cultured plant cells or protoplasts of those cells from the plant species disclosed herein. The experiments are performed using different eukaryotic cells including plant and mammalian cells, including, for example, 293 cells (transformed human primary kidney cells), HeLa cells (human cervical carcinoma), and H1299 (human epithelial carcinoma, non-small cell lung cancer). Hela cells are grown at 37°C and 5% CO, in a humidified incubator to a density of 2 x 10° cells/ml in an 8 chamber slide (Lab-Tek). After replacing the

28-12-2001 01939797 regular DMEM with Optimem, the cells are co-transfected with 10 pg of plasmid pAURNeo(-)FIAsH and pg of modified single-stranded oligonucleotide (35/25G) that is previously complexed with 10 ug lipofectamine, according to the manufacturer's directions (Life Technologies). The cells are treated with the liposome-DNA-oligo mix for 6 hrs at 37°C. Treated cells are washed with PBS and fresh DMEM is added. After a 16-18 hr recovery period, the culture is assayed for gene repair. The same oligonucleotide used in the cell-free extract experiments is used to target transfected plasmid bearing the kan® gene. Correction of the point mutation in this gene eliminates a stop codon and restores full expression. This expression can be detected by adding a small non-fluorescent ligand that bound to a

C-C-R-E-C-C sequence (SEQ ID NO: 2717) in the genetically modified carboxy terminus of the kan protein, to produce a highly fluorescent complex (FIAsH system, Aurora Biosciences Corporation). @® Following a 60 min incubation at room temperature with the ligand (FIAsH-EDT?2), cells expressing full length kan product acquire an intense green fluorescence detectable by fluorescence microscopy using a fluorescein filter set. Similar experiments are performed using the HygeGFP target as described in

Example 2 with a variety of mammalian cells, including, for example, COS-1 and COS-7 cells (African green monkey), and CHO-K1 cells (Chinese hamster ovary). The experiments are also performed with

PG12 cells (rat pheochromocytoma) and ES cells (human embryonic stem cells).

Summary of experimental results. Tables 1, 2 and 3 respectively provide data on the efficiency of gene repair directed by single-stranded oligonucleotides. Table 1 presents data using a cell- free extract from human liver cells (HUH?) to catalyze repair of the point mutation in plasmid pkan®m4021 (see Figure 1). Table 2 illustrates that the oligomers are not dependent on MSH2 or MSH3 for optimal gene repair activity. Table 3 illustrates data from the repair of a frameshift mutation (Figure 3) in the tet ® gene contained in plasmid pTetA208. Table 4 illustrates data from repair of the pkanm4021 point mutation catalyzed by plant cell extracts prepared from canola and musa (banana). Colony numbers are presented as kan" or tet’ and fold increases (single strand versus double hairpin) are presented for kan" in

Table 1.

Figure 5A is a confocal picture of Hel.a cells expressing the corrected fusion protein from an episomal target. Gene repair is accomplished by the action of a modified single-stranded oligonucleo- } tide containing 3 phosphorothioate linkages at each end (3S/25G). Figure 5B represents a "Z-series" of

Hela cells bearing the corrected fusion gene. This series sections the cells from bottom to top and illustrates that the fluorescent signal is "inside the cells".

Results. In summary, we have designed a novel class of single-stranded oligonucleotides with backbone modifications at the termini and demonstrate gene repair/conversion

AMENDED SHEET activity in mammalian and plant cell-free extracts. We confirm that the all DNA strand of the RNA-DNA double-stranded double hairpin chimera is the active component in the process of gene repair. In some . cases, the relative frequency of repair by the. novel oligonucleotides of the invention is elevated approximately 3-4-fold in certain embodiments when compared to frequencies directed by chimeric RNA-

DNA double hairpin oligonucleotides.

This strategy centers around the use of extracts from various sources to correct a mutation in a plasmid using a modified single-stranded or a chimeric RNA-DNA double hairpin oligonucleotide. A mutation is placed inside the coding region of a gene conferring antibiotic resistance in bacteria, here kanamycin or tetracycline. The appearance of resistance is measured by genetic readout in E.coli grown in the presence of the specified antibiotic. The importance of this system is that both phenotypic alteration and genetic inheritance can be measured. Plasmid pK®m4021 contains a mutation (T=3G) at residue 4021 rendering it unable to confer antibiotic resistance in E.coli. This point mutation is targeted for repair by oligonucleotides designed to restore kanamycin resistance. To avoid concerns of plasmid contamination skewing the colony counts, the directed correction is from G=#C rather than G=$T (wild-type). After isolation, the plasmid is electroporated into the DH10B strain of E.coli, which contains inactive RecA protein. The number of kanamycin colonies is counted and normalized by ascertaining the number of ampicillin colonies, a process that controls for the influence of electroporation. The number of colonies generated from three to five independent reactions was averaged and is presented for each experiment. A fold increase number is recorded to aid in comparison. * The original RNA-DNA double hairpin chimera design, e.g., as disclosed in U.S.

Patent 5,565,350, consists of two hybridized regions of a single-stranded oligonucleotide folded into a double hairpin configuration. The double-stranded targeting region is made up of a 5 base pair DNA/DNA segment bracketed by 10 base pair RNA/DNA segments. The central base pair is mismatched to the corresponding base pair in the target gene. When a molecule of this design is used to correct the kan® mutation, gene repair is observed (I in Figure 1A). Chimera li (Figure 1B) differs partly from chimera l in that only the DNA strand of the double hairpin is mismatched to the target sequence. When this chimera was used to correct the kan® mutation, it was twice as active. In the same study, repair function could be further increased by making the targeting region of the chimera a continuous RNA/DNA hybrid.

Frame shift mutations are repaired. By using plasmid pT°A208, described in Figure 1(C) and Figure 3, the capacity of the modified single-stranded molecules that showed activity in correcting a point mutation, can be tested for repair of a frameshift. To determine efficiency of correction of the mutation, a chimeric oligonucleotide (Tet I), which is designed to insert a T residue at position 208, is

28-12-2001 01939797 used. A modified single-stranded oligonucleotide (Tet IX) directs the insertion of a T residue at this same site. Figure 3 illustrates the plasmid and target bases designated for change in the experiments. When all reaction components are present (extract, plasmid, oligomer), tefracycline resistant colonies appear.

The colony count increases with the amount of oligonucleotide used up to a point beyond which the count } falls off (Table 3). No colonies above background are observed in the absence of either extract or oligonucleotide, nor when a modified single-stranded molecule bearing perfect complementarity is used.

Figure 3 represents the sequence surrounding the target site and shows that a T residue is inserted at the correct site. We have isolated plasmids from fifteen colonies obtained in three independent experiments and each analyzed sequence revealed the same precise nucleotide insertion. These data suggest that the single-stranded molecules used initially for point mutation correction can also repair 9 nucleotide deletions.

Comparison of phosphorothioate oligonucleotides to 2'-O-methyl substituted oligonucleotides. From a comparison of molecules VII and XI, it is apparent that gene repair is more subject to inhibition by RNA residues than by phosphorothioate linkages. Thus, even though both of these oligonucleotides contain an equal number of modifications to impart nuclease resistance, X| (with 16 phosphorothioate linkages) has good gene repair activity while VII (with 16 2'-O-methyl RNA residues) is inactive. Hence, the original chimeric double hairpin oligonucleotide enabled correction directed, in large part, by the strand containing a large region of contiguous DNA residues.

Oligonucleotides can target multiple nucleotide alterations within the same template.

The ability of individual single-stranded oligonucleotides to correct multiple mutations in a single target template is tested using the plasmid pK’m4021 and the following single-stranded oligonucleotides ® modified with 3 phosphorothioate linkages at each end (indicated as underlined nucleotides): Oligo1 is a 25-mer with the sequence TICGATAAGCCTATGCTGACCCGTG (SEQ ID NO: 2709) corrects the original mutation present in the kanamycin resistance gene of pK’m4021 as well as directing another alteration 2 basepairs away in the target sequence (both indicated in boldface); Oligo2 is a 70-mer with the 5-end sequence TTCGGCTACGACTGGGCACAACAGACAATTGGC (SEQ ID NO: 2710) with the remaining nucleotides being completely complementary to the kanamycin resistance gene and also ending in 3 phosphorothioate linkages at the 3' end. Oligo2 directs correction of the mutation in pK'm4021 as well as directing another alteration 21 basepairs away in the target sequence (both indicated in boldface).

We also use additional oligonucleotides to assay the ability of individual oligonucleotides to correct multiple mutations in the pK"M4021 plasmid. These include, for example, a second 25-mer that alters two nucleotides that are three nucleotides apart with the sequence 5'-

AMENDED SHEET

28-12-2001 01939797

TTGTGCCCAGTCGTATCCGAATAGC-3' (SEQ ID NO: 2711); a 70-mer that alters two nucleotides that are 21 nucleotides apart with the sequence 5'-CATCAGAGCAGCCAATTGTCTGTTGTGCCCAGTCGTA

GCCGAATAGCCTCTCCACCCAAGCGGCCGGAGA-3 (SEQ ID NO: 2712); and another 70-mer that alters two nucleotides that are 21 nucleotides apart with the sequence 5-GCTGACAGCCGGAACACGG

CGGCATCAGAGCAGCCAATTGTCTGTTGTGCCCAGTCGTAGCCGAATAGCCT-3' (SEQ ID NO: 2713).

The nucleotides in the oligonucleotides that direct alteration of the target sequence are underlined and in boldface. These oligonucleotides are modified in the same way as the other oligonucleotides of the invention.

We assay correction of the original mutation in pK*m4021 by monitoring kanamycin resistance (the second alterations which are directed by Oligo2 and Qligo3 are silent with respect to the ® kanamycin resistance phenotype). In addition, in experiments with Oligo2, we also monitor cleavage of the resulting plasmids using the restriction enzyme Tsp5091 which cuts at a specific site present only when the second alteration has occurred (at ATT in Oligo2). We then sequence these clones to determine whether the additional, silent alteration has also been introduced. The resulls of an analysis are presented below: oigoiesmen | Oigo2(omen

Cuero |» | 7

Cloneswihasngesiechanged | 0 | 2 ® Nuclease sensitivity of unmodified DNA oligonucleotide. Electrophoretic analysis of nucleic acid recovered from the cell-free extract reactions conducted here confirm that the unmodified single-stranded 25-mer did not survive incubation whereas greater than 90% of the terminally modified oligos did survive (as judged by photo-image analyses of agarose gels).

Plant extracts direct repair. The modified single-stranded constructs can be tested in plant cell extracts. We have observed gene alteration using extracts from multiple plant sources, including, for example, Arabidopsis, tobacco, banana, maize, soybean, canola, wheat, spinach as well as spinach chloroplast extract or extracts made from other plant cells disclosed herein. We prepare the extracts by grinding plant tissue or cultured cells under liquid nitrogen with a mortar and pestle. We extract 3 ml of the ground plant tissue with 1.5 ml of extraction buffer (20 mM HEPES, pH7.5; 5 mM KCI;

AMENDED SHEET

1.5 mM MgCl,; 10 mM DTT; and 10% [v/v] glycerol). Some plant cell-free extracts also include about 1% (wiv) PVP. We then homogenize the samples with 15 strokes of a Dounce homogenizer. Following homogenization, we incubate the samples on ice for 1 hour and centrifuge at 3000 x g for 5 minutes to remove plant cell debris. We then determine the protein concentration in the supernatants (extracts) by : 5 Bradford assay. We dispense 100 ig (protein) aliquots of the extracts which we freeze in a dry ice- ethanol! bath and store at -80°C.

We describe experiments using two sources here: a dicot (canola) and a monocot (banana, Musa acuminata cv. Rasthali). Each vector directs gene repair of the kanamycin mutation (Table 4); however, the level of correction is elevated 2-3 fold relative to the frequency observed with the chimeric oligonucleotide. These results are similar to those observed in the mammalian system wherein a significant improvement in gene repair occurred when modified single-stranded molecules were used.

Tables are attached hereto.

: Table I

Gene repair activity is directed by single-stranded oligonucleotides.

Oligonucleotide Plasmid Extract (ug) kan’ colopies Fold increase

I pK°m4021 10 300

I 20 418 1.0x 1 10 537 il 20 748 1.78x m 10 3 m 20 5 0.01x

Iv 10 112

Iv 20 96 0.22x \'% 10 217

AY 20 342 0.81x

VI 10 6

A 20 39 0.093x vi 10 0 vi 20 0 0x via 10 3 vi 20 5 0.01x

IX 10 936

IX 20 1295 3.09x

X 10 1140

X 20 1588 3.7x

Xi 10 480

XI 20 681 1.6x

Xn 10 18

XI 20 25 0.059x

Xm 10 0

Xm 20 4 0.009x . 20 0 :

I - 1

Plasmid pK'm4021 (1pg), the indicated oligonucleotide (1.5 ug chimeric oligonucleotide or 0.55 pg single-stranded oligonucleotide; molar ratio of oligo to plasmid of 360 to 1) and either 10 or 20 pug of HUH? cell-free extract were incubated 45 min at 37°C. Isolated plasmid DNA was electroporated into E. coli (strain DH10B) and the number of kar’ - colonies counted. The data represent the number of kanamycin resistant colonies per 10° ampicillin resistant colonies generated from the same reaction and is the average of three experiments (standard deviation usually less than +/- 15%). Fold increase is defined relative to 418 kan’ colonies (second reaction) and in all reactions was calculated using the 20pug sample.

Table II

Modified single-stranded oligomers are not dependent on MSH2 or MSHS3 for optimal gene repair activity.

A. Oligonucleotide Plasmid Extract "colonies

IX (358/25G) HUH7 637

X (6S/25G) HUH7 836

X MEF2™ 781

X MEF2™ 676

IX MEF3™ 582

X MEF3™ 530 © IX MEF™ 332

X MEF" 497 . MEF2™ 10 - MEF3™ 5 oo MEF 14

Chimeric oligonucleotide (1.5 ug) or modified single-stranded oligonucleotide (0.55 pg) was incubated with 1g of plasmid pK'm4021 and 20ug of the indicated extracts. MEF represents mouse embryonic fibroblasts with either MSH2 (27) or MSH3 (3*) deleted.

MEF" indicates wild-type mouse embryonic fibroblasts. The other reaction components were then added and processed through the bacterial readout system. The data represent the number of kanamycin resistant colonies per 10° ampicillin resistant colonies.

Table III

Frameshift mutation repair is directed by single-stranded oligonucleotides

Oligonucleotide Plasmid Extract tet” colonies

Tet IX (38/25A;0.5 ug) - pT*A208 (lpg) - 0 - 20pg 0

Tet IX (0.5 pg) 48 :

Tet IX (1.5 pg) 130

Tet IX (2.0 pg) 68

Tet I (chimera; 1.5 pg) 48

Each reaction mixture contained the indicated amounts of plasmid and oligonucleotide. :

The extract used for these experiments came from HUH? cells. The data represent the number of tetracycline resistant colonies per 10° ampicillin resistant colonies generated from the same reaction and is the average of 3 independent experiments. TetIisa chimeric oligonucleotide and Tet IX is a modified single-stranded oligonucleotide that are designed to insert a T residue at position 208 of pT*A208. These oligonucleotides are equivalent to structures I and IX in Figure 2.

Table IV

Plant cell-free. extracts support gene repair by single-stranded oligonucleotides

Oligonucleotide Plasmid Extract kan’ colonies

II (chimera) pK°m4021 30g Canola 337

IX (38/25G) Canola 763

X (65/25G) Canola 882 1 : Musa 203

IX Musa - 343

X Musa 746 - Canola 0 - Musa 0

IX - Canola 0

X - Musa 0

Canola or Musa cell-free extracts were tested for gene repair activity on the kanamycin- sensitive gene as previously described in (18). Chimeric oligonucleotide IT (1.5 pg) and modified single-stranded oligonucleotides IX and X (0.55ug) were used to correct pKSth4021. Total number of kan" colonies are present per 107 ampicillin resistant colonies and represent an average of four independent experiments.

: § 2=S%sn i: - OO rem gg

PE 5: : 5 § & ; g § 2 2 2 3 y _ 2 g ny § 3

BEE EE. il a KO. £ : ig ; 855 ig E—— Fig a 2 15 = 8 & a 8

Pd pea. 233 ~ Nn 0] t 4 s2<388 35 § 3 223354 2

EXAMPLE 2

Yeast Cell Targeting Assay Method for Base

Alteration and Preferred Oligonucleotide Selection

In this example, single-stranded oligonucleotides with modified backbones and double- * 5 hairpin oligonucleotides with chimeric, RNA-DNA backbones are used to measure gene repair using two episomal targets with a fusion between a hygromycin resistance gene and eGFP as a target for gene repair. These plasmids are pAURHYG(rep)GFP, which contains a point mutation in the hygromycin resistance gene (Figure 7), pAURHYG(ins)GFP, which contains a single-base insertion in the hygromycin resistance gene (Figure 7) and pAURHYG(A)GFP which has a single base deletion. We also use the plasmid containing a wild-type copy of the hygromycin-eGFP fusion gene, designated pAURHYG(wt)GFP, as a control. These plasmids also contain an aureobasidinA resistance gene. In pAURHYG(rep)GFP, hygromycin resistance gene function and green fluorescence from the eGFP protein are restored when a G at position 137, at codon 46 of the hygromycin B coding sequence, is converted to a C thus removing a premature stop codon in the hygromycin resistance gene coding region. In pAURHYG(ins)GFP, hygromycin resistance gene function and green fluorescence from the eGFP protein are restored when an A inserted between nucleotide positions 136 and 137, at codon 46 of the hygromycin B coding sequence, is deleted and a C is substituted for the T at position 137, thus correcting a frameshift mutation and restoring the reading frame of the hygromycin-eGFP fusion gene.

We synthesize the set of three yeast expression constructs pAURHYG(rep)eGFP, pAURHYG(A)eGFP, pAURHYG(ins)eGFP, that contain a point mutation at nucleotide 137 of the hygromycin-B coding sequence as follows. (rep) indicates a T137=2G replacement, (A) represents a deletion of the G137 and (ins) represents an A insertion between nucleotides 136 and 137. We construct this set of plasmids by excising the respective expression cassettes by restriction digest from pHyg(x)EGFP and ligation into pAUR123 (Panvera, CA). We digest 10 ug pAUR123 vector DNA, as well as, 10 ug of each pHyg(x)EGFP construct with Kpnl and Sali (NEB). We gel purify each of the DNA fragments and prepare them for enzymatic ligation. We ligate each mutated insert into pHygEGFP vector at 3:1 molar ratio using T4 DNA ligase (Roche). We screen clones by restriction digest, confirm by

Sanger dideoxy chain termination sequencing and purify using a Qiagen maxiprep kit.

We use this system to assay the ability of five oligonucleotides (shown in Figure 8) to support correction under a variety of conditions. The oligonucleotides which direct correction of the mutation in pAURHYG(rep)GFP can also direct correction of the mutation in pAURHYG(ins)GFP. Three of the four oligonucleotides (HygE3T/25, HygE3T/74 and HygGG/Rev) share the same 25-base sequence surrounding the base targeted for alteration. HygGG/Rev is an RNA-DNA chimeric double hairpin oligonucleotide of the type described in the prior art. One of these oligonucleotides, HygE3T/74, is a 74- base oligonucleotide with the 25-base sequence centrally positioned. The fourth oligonucleotide, ' designated HygE3T/74a, is the reverse complement of HygE3T/74. The fifth oligonucleotide, designated

Kan70T, is a non-specific, control oligonucleotide which is not complementary to the target sequence.

Alternatively, an oligonucleotide of identical sequence but lacking a mismatch to the target or a completely thioate modified oligonucleotide or a completely 2-0-methylated modified oligonucleotide may be used as a control. Alternatively, oligonucleotides containing one, two, three, four, five, six, eight, ten or more LNA modifications on at least one of the two termini (and preferrably the 3' terminus) may be used in different embodiments.

Oligonucleotide synthesis and cells. We synthesized and purified the chimeric, double- hairpin oligonucleotides and single-stranded oligonucleotides (including those with the indicated modifications) as described in Example 1. Plasmids used for assay were maintained stably in yeast (Saccharomyces cerevisiae) strain LSY678 MAT ar at low copy number under aureobasidin selection.

Plasmids and oligonucleotides are introduced into yeast cells by electroporation as follows: to prepare electrocompetent yeast cells, we inoculate 10 ml of YPD media from a single colony and grow the cultures overnight with shaking at 300 rpm at 30°C. We then add 30 ml of fresh YPD media to the overnight cultures and continue shaking at 30°C until the OD, was between 0.5 and 1.0 (3-5 hours). We then wash the cells by centrifuging at 4°C at 3000 rpm for 5 minutes and twice resuspending the cells in 25 ml ice-cold distilled water. We then centrifuge at 4°C at 3000 rpm for 5 minutes and resuspend in 1 ml ice-cold 1M sorbitol and then finally centrifuge the cells at 4°C at 5000 rpm for 5 minutes and resuspend the cells in 120 pi 1M sorbitol. To transform efectrocompetent cells with plasmids or oligonucleotides, we mix 40 ul of cells with 5 pg of nucleic acid, unless otherwise stated, and incubate on ice for 5 minutes.

We then transfer the mixture to a 0.2 cm electroporation cuvette and electroporate with a BIO-RAD Gene

Pulser apparatus at 1.5 kV, 25 pF, 200 Q for one five-second pulse. We then immediately resuspend the cells in 1 ml YPD supplemented with 1M sorbitol and incubate the cultures at 30°C with shaking at 300 rpm for 6 hours. We then spread 200 pil of this culture on selective plates containing 300 pg/ml hygromycin and spread 200 pi of a 10° dilution of this culture on selective plates containing 500 ng/ml aureobasidinA and/or and incubate at 30°C for 3 days to allow individual yeast colonies to grow. We then : count the colonies on the plates and calculate the gene conversion efficiency by determining the number of hygromycin resistance colonies per 10° aureobasidinA resistant colonies.

Frameshift mutations are repaired in yeast cells. We test the ability of the oligonucleotides shown in Figure 8 to correct a frameshift mutation in vivo using LSY678 yeast cells

28-12-2001 01939797 containing the plasmid pAURHYG(ins)GFP. These experiments, presented in Table 6, indicate that these oligonucleotides can support gene correction in yeast cells. These data reinforce the results described in Example 1 indicating that oligonucleotides comprising phosphorothioate linkages facilitate gene correction much more efficiently than control duplex, chimeric RNA-DNA oligonucleotides. This gene correction activity is also specific as transformation of cells with the control oligonucleotide Kan70T produced no hygromycin resistant colonies above background and thus Kan70T did not support gene correction in this system. In addition, we observe that the 74-base oligonucleotide (HygE3T/74) corrects the mutation in pAURHYG(ins)GFP approximately five-fold more efficiently than the 25-base oligonucleotide (HygE3T/25). We also perform control experiments with LSY678 yeast cells containing the plasmid pAURHYG(wt)GFP. With this strain we observed that even without added oligonucleotides, ® there are too many hygromycin resistant colonies to count.

We also use additional oligonucleotides to assay the ability of individual oligonucleotides to correct multiple mutations in the pAURHYG(x)eGFP plasmid. These include, for example, one that alters two basepairs that are 3 nucleotides apart is a 74-mer with the sequence 5-CTCGTGCTTTCAGCT

TCGATGTAGGAGGGCGTGGGTACGTCCTGCGGGTAAATAGCTGCGCCGATGGTTTCTAC-3' (SEQ ID

NO: 2714); a 74-mer that alters two basepairs that are 15 nucleotides apart with the sequence 5

CTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGATACGTCCTGCGGGTAAACAGCTGCGCCGATG

GTTTCTAC-3' (SEQ ID NO: 2715); and a 74-mer that alters two basepairs that are 27 nucleotides apart with the sequence 5-CTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGATACGTCCTGCGGGTA

AATAGCTGCGCCGACGGTTTCTAC (SEQ ID NO: 2716). The nucleotides in these oligonucleotides that direct alteration of the target sequence are underlined and in boldface. These oligonucleotides are ® modified in the same ways as the other oligonucleotides of the invention.

Oligonucleotides targeting the sense strand direct gene correction more efficiently. We compare the ability of single-stranded oligonucleotides to target each of the two strands of the target sequence of both pAURHYG(ins)GFP and pAURHYG(rep)GFP. These experiments, presented in Tables 7 and 8, indicate that an oligonucleotide, HygE3T/74cx, with sequence complementary to the sense strand (i.e. the strand of the target sequence that is identical to the mRNA) of the target sequence : facilitates gene correction approximately ten-fold more efficiently than an oligonucleotide, HygE3T/74, with sequence complementary to the non-transcribed strand which serves as the template for the synthesis of RNA. As indicated in Table 7, this effect was observed over a range of oligonucleotide concentrations from 0-3.6 Hg, although we did observe some variability in the difference between the two oligonucleotides (indicated in Table 7 as a fold difference between HygE3T/74x and HygE3T/74).

AMENDED SHEET

Furthermore, as shown in Table 8, we observe increased efficiency of correction by HygE3T/74ax relative to HygE3T/74 regardless of whether the oligonucleotides were used to correct the base substitution : mutation in pAURHYG(rep)GFP or the insertion mutation in pAURHYG(ins)GFP. The data presented in

Table 8 further indicate that the single-stranded oligonucleotides correct a base substitution mutation "5 more efficiently than an insertion mutation. However, this last effect was much less pronounced and the oligonucleotides of the invention are clearly able efficiently to correct both types of mutations in yeast cells. In addition, the role of transcription is investigated using plasmids with inducible promoters such as that described in Figure 10.

Optimization of oligonucleotide concentration. To determine the optimal concentration of - oligonucleotide for the purpose of gene alteration, we test the ability of increasing concentrations of

Hyg3T/74x to correct the mutation in pAURHYG(rep)GFP contained in yeast LSY678. We chose this assay system because our previous experiments indicated that it supports the highest level of correction.

However, this same approach could be used to determine the optimal concentration of any given oligonucleotide. We test the ability of Hyg3T/74ax to correct the mutation in pAURHYG(rep)GFP contained in yeast LSY678 over a range of oligonucleotide concentrations from 0-10.0 ug. As shown in

Table 9, we observe that the correction efficiency initially increases with increasing oligonucleotide concentration, but then declines at the highest concentration tested.

Tables are attached hereto.

Table 6

Correction of an insertion mutation in pAURHYG(ins)GFP by HygGG/Rev, HygE3T/25 and HygE3T/74 . Oligonucleotide Tested | Colonies on Colonies on Correction

Hygromycin Aureobasidin (/10°) | Efficiency

HygGG Rey

HyGEST/74

Kan?OT HE

Table 7

An oligonucleotide targeting the sense strand of the target sequence corrects more efficiently.

Amount of Oligonucleotide (ug) Colonies per hygromycin plate ww meer

EET YT

167 (5.8) 367 (15%) * The numbers in parentheses represent the fold increase in efficiency for targeting the non-transcribed strand as compared to the other strand of a DNA duplex that encodes a protein.

Table 8

Correction of a base substitution mutation is more efficient than correction of a frame shift mutation. ; = ET

Table 9

Optimization of oligonucleotide concentration in electroporated yeast cells.

Amount (Mg) Colonies on Colonies on Correction efficiency aa Fee Fe i]

II ECR DT 2 1

Example 3

Cultured Cell Manipulation

Although disclosure in this example is directed to use of stem cells or human blood cells and microinjection, the microinjection procedures may also be used with cultured plant cells or protoplasts using any plant species, including those disclosed herein. Mononuclear cells are isolated from human umbilical cord blood of normal donors using Ficoll Hypaque (Pharmacia Biotech, Uppsala, Sweden) density centrifugation. CD34+ cells are immunomagnetically purified from mononuclear cells using either the progenitor or Multisort Kits (Miltenyi Biotec, Auburn, CA). Lin"CD38" cells are purified from the mononuclear cells using negative selection with StemSep system according to the manufacturer's protocol (Stem Cell Technologies, Vancouver, CA). Cells used for microinjection are either freshly isolated or cryopreserved and cultured in Stem Medium (S Medium) for 2 to 5 days prior to microinjection.

S Medium contains Iscoves’ Modified Dulbecco’s Medium without phenol red (IMDM) with 100 pg/ml glutamine/penicillin/streptomycin, 50 mg/ml bovine serum albumin, 50 ug/ml bovine pancreatic insulin, 1 mg/m! human transferrin, and IMDM; Stem Cell Technologies), 40 pg/ml low-density lipoprotein (LDL;

Sigma, St. Louis, MO), 50 mM HEPEs buffer and 50 uM 2-mercaptoethanol, 20 ng/ml each of thrombopoietin, fit-3 ligand, stem cell factor and human IL-6 (Pepro Tech Inc., Rocky Hill, NJ). After microinjection, cells are detached and transferred in bulk into wells of 48 well plates for culturing. 35 mm dishes are coated overnight at 4° C with 50 pg/ml Fibronectin (FN) fragment Ch- 296 (Retronectin; TaKaRa Biomedicals, Panvera, Madison, W) in phosphate buffered saline and washed with IMDM containing glutamine/penicillin/streptomycin. 300 to 2000 cells are added to cloning rings and attached to the plates for 45 minutes at 37° C prior to microinjection. After incubation, cloning rings are removed and 2 mi of S Medium are added to each dish for microinjection. Pulled injection needles with a range of 0.22 um to 0.3 um outer tip diameter are used. Cells are visualized with a microscope equipped with a temperature controlled stage set at 37° C and injected using an electronically interfaced Eppendorf

Micromanipulator and Transjector. Successfully injected cells are intact, alive and remain attached to the plate post injection. Molecules that are flourescently labeled allow determination of the amount of oligonucleotide delivered to the cells.

For in vitro erythropoiesis from Lin"CD38" cells, the procedure of Malik, 1998 can be used. Cells are cultured in ME Medium for 4 days and then cultured in E Medium for 3 weeks.

Erythropoiesis is evident by glycophorin A expression as well as the presence of red color representing the presence of hemoglobin in the cultured cells. The injected cells are able to retain their proliferative capacity and the ability to generate myeloid and erythoid progeny. CD34+ cells can convert a normal A (B") to sickle T (B°) mutation in the B-globin gene or can be altered using any of the oligonucleotides of } the invention herein for correction or alteration of a normal gene to a mutant gene. Alternatively, stem cells can be isolated from blood of humans having genetic disease mutations and the oligonucleotides of : the invention can be used to correct a defect or to modify genomes within those cells.

Alternatively, non-stem cell populations of cultured cells can be manipulated using any method known to those of skill in the art including, for example, the use of polycations, cationic lipids,

liposomes, polyethylenimine (PEI), electroporation, biolistics, calcium phosphate precipitation, or any other method known in the art.

Biolistic delivery of oligonucleotide into plant cells may be accomplished according to the following method. One milliliter of packed cell volume of plant cell suspensions are subcultured onto plates containing solid medium [with Murashige and Skoog salts from Gibco/BRL, 500 mg/liter Mes, 1 mglliter thiamin, 100 mg/liter myo-inositol, 180 mg/liter KH2PO4, 2.21 mglliter 2,4-dichlorophenoxyacetic acid (2,4-D), and 30 g/liter sucrose (pH 5.7) and having 8 g/liter agar-agar from Sigma added before autoclaving]. By using a helium-driven particle gun such as that from BioRad and following manufacturers directions, oligonucleotides may be introduced to cells after precipitation onto 1 micrometer or comparable gold microcarriers (Bio-Rad). To precipitate onto microcarriers, 35 microliters of a particle suspension (60 mg of microcarriers per mi of 100% ethanol) is transferred to a 1.5 ml microcentrifuge tube, which is agitated on a vortex mixer. Then 40 microliter of resuspended oligonucleotide (60 ng/microliter water) is added; then 75 microliter of ice-cold 2.5 M CaCl2 is added; then 75 microliter of ice- cold 0.1 M spermidine is added. The tube is mixed vigorously or a vortex mixer for 10 min at room temperature. The particles are allowed to settle for 10 min and are centrifuged at 11,750 g for 30 sec.

The supernatant is removed and the particles are resuspended in 50 microliter of 100% ethanol. An aliquot of 10 microliter of the resuspended particles are applied to each macro-projectile which is used to bombard each plate once at 900 psi (1 psi = 6.89 kPa) with a gap distance (distance from power source to macroprojectile) of 1 cm and a target distance (distance from microprojectile launch site to target material) of 10 cm.

An alternative method of delivery can be used as follows. Cultured cells are suspended in liquid

N6 medium and then plated on a VWR Scientific glass fiber filter. About 0.4 microgram of oligonucleotide are precipitated with 15 microliter of 2.5 mM CaCI2 and 5 microliter of 0.1 M spermidine onto 25 microgram of 1.0 micrometer gold particles. Microprojectile bombardment is performed by using a Bio-

Rad PDS-1000 He particle delivery system or comparable machine following manufacturers instructions.

Alterations in oligonucleotide concentrations can be employed to determine the optimum concentration of oligonucleotide according to the procedures described herein for any particular oligonucleotide of the invention. : Alternatively, the oligonucleotide of the invention may be delivered to a plant cell by electroporation of a protoplast derived from a plant part. The protoplasts may be formed by enzymatic treatment of a plant part, particularly a leaf, according to techniques such as those in Gallois et al.,

Methods in Molecular Biology 55: 89-107 by Humana Press. Such conditions for electroporation use about 3 x 10° protoplasts in a total volume of about 0.3 ml with a concentration of oligonucleotide of between 0.6 to 4 microgram per ml.

EXAMPLE 4

Plant Cells

The oligonucleotides of the invention can also be used to repair or direct a mutagenic event in plants and animal cells. Although little information is available on plant mutations amongst : natural cultivars, the oligonucleotides of the invention can be used to produce "knock out" mutations by modification of specific amino acid codons to produce stop codons (e.g., a CAA codon specifying Gin can be modified at a specific site to TAA; a AAG codon specifying Lys can be modified to UAG at a specific site; and a CGA codon for Arg can be modified to a UGA codon at a specific site). Such base pair changes will terminate the reading frame and produce a defective truncated protein, shortened at the site of the stop codon . Alternatively, frameshift additions or deletions can be directed into the genome at a specific sequence to interrupt the reading frame and produce a garbled downstream protein. Such stop or frameshift mutations can be introduced to determine the effect of knocking out the protein in either plant or animal cells.

For introduction of a T-DNA, including the T-DNA in the plasmid of Figure 11, into a plant cell, Agrobacterium tumefaciens is used. These techniques are routine standard techniques known in the art. For example, one method follows. We transform A. tumefaciens is transformed by electroporation (using a BioRad Gene Pulser"). Competent A. tumefaciens is prepared using a method similar to that of preparing competent E. coli by suspending a freshly grown culture three times in ice-cold water and a final resuspension in 10% glycerol. Electroporation conditions are a 0.2 cm gap cuvette at a setting of 25

WF, 200 Q and 2.5 kV.

A. tumefaciens containing a plasmid with a T-DNA is then used to introduce the T-DNA into a plant cell using routine standard techniques known in the art. For example, we transform )

Arabidopsis by vacuum infiltration or by dipping flowers in an Agrobacterium solution containing a surfactant, e.g. L-77. Seeds are then collected, grown and screened for presence of the T-DNA. ) Alternatively, Agrobacterium can be used to transform callus tissue and the callus tissue can then be . used to regenerate transformed plants.

All publications and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and } modifications may be made thereto without departing from the spirit or scope of the appended claims.

Notes on the tables presented below: oo

Each of the following tables presents, for the specified gene, a plurality of mutations that are known to confer a relevant phenotype and, for each mutation, the oligonucleotides that can be used to correct the respective mutation site-specifically in the genome according to the present invention.

The left-most column identifies each alteration or mutation and the phenotype that the alteration/mutation confers.

For most entries, the mutation/alteration is identified at both the nucleic acid and protein level. At the amino acid level, mutations are presented according to the following standard nomenclature. : The centered number identifies the position of the mutated codon in the protein sequence; to the left of the number is the wild type residue and to the right of the number is the mutant codon. Terminator codons are shown as "TERM". At the nucleic acid level, the entire triplet of the wild type and mutated codons is shown.

The middie column presents, for each mutation, four oligonucleotides capable of repairing the mutation site-specifically in the genome or in cloned DNA including DNA in artificial chromosomes, episomes, plasmids, or other types of vectors. The oligonucleotides of the invention, however, may include any of the oligonucleotides sharing portions of the sequence of the 121 base sequence. Thus, oligonucleotides of the invention for each of the depicted targets may be 18, 19, 20 up to about 121 nucleotides in length. Sequence may be added non-symmetrically.

All oligonucleotides are presented, per convention, in the 5' to 3' orientation. The nucleotide that effects the change in the genome is underlined and presented in bold.

The first of the four oligonucleotides for each mutation is a 121 nt oligonucleotide centered about the repair/altering nucleotide. The second oligonucleotide, its reverse complement, ) targets the opposite strand of the DNA duplex for repair/alteration. The third oligonucleotide is the minimal 17 nt domain of the first oligonucleotide, also centered about the repair/alteration nucleotide. The : fourth oligonucleotide is the reverse complement of the third, and thus represents the minimal 17 nt domain of the second.

The third column of each table presents the SEQ ID NO: of the respective repair oligonucleotide.

Example 5

Engineering herbicide resistant plants

Chemical weed control is an important tool of modern agriculture and many herbicides have been developed for this purpose. Their use has resulted in substantial increases in the yields of many crops, including, for example, maize, soybeans, and cotton. Thus while the use of fertilizers and new high-yielding crop varieties have contributed greatly to the “green revolution," chemical weed control has also been at the forefront of technological achievement.

Herbicides having broad-spectrum activity are particularly useful because they obviate the need for multiple herbicides targeting different classes of weeds. The problem with such herbicides is that they typically also affect crops which are exposed fo the herbicide. One way to overcome this is to generate plants which are resistant to one or more broad-spectrum herbicides. Such herbicide-tolerant plants may reduce the need for tillage to control weeds, thereby effectively reducing soil erosion and can reduce the quantity and number of different herbicides applied in the field.

Common herbicides used, for example, include those that inhibit the enzyme 5-enolpyruvyl-3-phosphoshikimic acid synthase (EPSPS), for example N-phosphonomethyi-glycine (e.g. glyphosate), those that inhibit acetolactate synthase (ALS) activity, for example the sulfonylureas and related herbicides, and those that inhibit dihydropteroate synthase, for example methyl[(4-amino- phenyl)sulfonyljcarbamate (e.g. Asulam). Herbicide-tolerant plants can be produced by several methods, including, for example, introducing into the genome of the plant the ability to degrade the herbicide, the capacity to produce a higher level of the targeted enzyme, and/or expressing an herbicide-tolerant allele of the enzyme.

The attached tables disclose exemplary oligonucleotides base sequences which can be used to generate site-specific mutations in plant genes that confer herbicide resistance.

28-12-2001 01939797

Table 10

Genome-Alfering Oligos Conferring Glyphosate Resistance

Phenotype, Gene, . Plant & Targeted .

Alteration Altering Oligos 0:

Clyphosate Resistance | AAGCGTCGGAGATTGTACTTCAACCCATTAGAGAAATCTCCGGTC

EPSPS TTATTAAGCTTCCTGCCTCCAAGTCTCTATCAAATCGGATCCTGC

Arabidopsis thaliana TTCTCGCTGCTCTGTCTGAGGTATATATCAC

Gly97Ala GTGATATATACCTCAGACAGAGCAGCGAGAAGCAGGATCCGATT 2

GGC-GCC TGATAGAGACTTGGAGGCAGGAAGCTTAATAAGACCGGAGATTT

CTCTAATGGGTTGAAGTACAATCTCCGACGCTT

® GCTTCCTGCCTCCAAGT

ACTTGGAGGCAGGAAGC slyphosate Resistance [AAGCTTCAGAGATTGTGCT T1CAACCAATCAGAGAAATCTCGGGT 5

EPSPS TCATTAAGCTACCCGCATCCAAATCTCTCTCCAATCGGATCCTCC

Brassica napus TTCTTGCCGCTCTATCTGAGGTACATATACT

Gly93Ala AGTATATGTACCTCAGATAGAGCGGCAAGAAGGAGGATCCGATT

GGA-GCA GGAGAGAGATTTGGATGCGGGTAGCTTAATGAGACCCGAGATTT

CTCTGATTGGTTGAAGCACAATCTCTGAAGCTT

GCTACCCGCATCCAAAT

ATTTGGATGCGGGTAGC | 8

Glyphosate Resistance | AGCCCAACGAGATTGTGCTGCAACCCATCAAAGATATATCAGGC

EPSPS 1 ACTGTTAAATTGCCTGCTTCTAAATCCCTTTCCAATCGTATTCTCC

Nicotiana tabacum TTCTTGCTGCCCTTTCTAAGGGAAGGACTGT

Gly9bAla ACAGTCCTTCCCTTAGAAAGGGCAGCAAGAAGGAGAATACGATT 10

GGT-GCT GGAAAGGGATTTAGAAGCAGGCAATTTAACAGTGCCTGATATATC ® TTTGATGGGTTGCAGCACAATCTCGTTGGGCT

ATTGCCTGCTTCTAAAT

ATTTAGAAGCAGGCAAT

Glyphosate Resistance | ATTGTTTCCTTGGTACGAAATGTCCTCCTGITCGAATIGTCAGCA 3

EPSPS 2 AGGGAGGCCTTCCCGCAGGGAAGGTAAAGCTCTCTGGATCAATT

Nicotiana tabacum AGCAGCCAGTACTTGACTGCTCTGCTTATGGC

Gly62Ala GCCATAAGCAGAGCAGTCAAGTACTGGCTGCTAATTGATCCAGA 14

GGA-GCA GAGCTTTACCTTCCCTGCGGGAAGGCCTCCCTTGCTGACAATTC

GAACAGGAGGACATTTCGTACCAAGGAAACAAT

CCTTCCCGCAGGGAAGG

CCTTCCCTGCGGGAAGG yphosate Resistance | ATTGTTTCCTTGGCACTGACTGCCCACCTGTTCGTGTCAATGGAA 17 : EPSPS TCGGAGGGCTACCTGCTGGCAAGGTCAAGCTGTCTGGCTCCATC

Zea mays AGCAGTCAGTACTTGAGTGCCTTGCTGATGGC

Gly168Ala GCCATCAGCAAGGCACTCAAGTACTGACTGCTGATGGAGCCAGA | 18

GGT-GCT CAGCTTGACCTTGCCAGCAGGTAGCCCTCCGATTCCATTGACAC

GAACAGGTGGGCAGTCAGTGCCAAGGAAACAAT

1112/01 01:47 pm

AMENDED SHEET 3137.009 — [NY]793559.1

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted . . : Alteration Altering Oligos

GCTACCTGCTGGCAAGG

CCTTGCCAGCAGGTAGC

Glyphosate Resistance [ACTGTTTCCTTGGCACTGAATGCCCACCTGTTCGTGTCAAGGGA 2

EPSPS ATTGGAGGACTTCCTGCTGGCAAGGTTAAGCTCTCTGGTTCCAT

Oryza sativa CAGCAGTCAGTACTTGAGTGCCTTGCTGATGGC

Gly115Ala GCCATCAGCAAGGCACTCAAGTACTGACTGCTGATGGAACCAGA | 22

GGT-GCT GAGCTTAACCTTGCCAGCAGGAAGTCCTCCAATTCCCTTGACAC

GAACAGGTGGGCATTCAGTGCCAAGGAAACAGT

ACTTCCTGCTGGCAAGG

® CCTTGCCAGCAGGAAGT yphosate Resistance | AGCCTTCTGAGATAGTGTTGCAACCCATTAAAGAGATTTCAGGCA 5

EPSPS CTGTTAAATTGCCTGCCTCTAAATCATTATCTAATAGAATTCTCCT

Petunia x hybrida TCTTGCTGCCTTATCTGAAGGAACAACTGT

Gly93Ala ACAGTTGTTCCTTCAGATAAGGCAGCAAGAAGGAGAATTCTATTA | 26

GGC-GCC GATAATGATTTAGAGGCAGGCAATTTAACAGTGCCTGAAATCTCT

TTAATGGGTTGCAACACTATCTCAGAAGGCT

ATTGCCTGCCTCTAAAT

ATTTAGAGGCAGGCAAT

[Glyphosate Resistance | AACCCCATGAGATTGTGCTAGNACCCATCAAAGATATATCTGGTA | 29

EPSPS CTGTTAAATTACCCGCTTCGAAATCCCTTTCCAATCGTATTCTCCT :

Lycopersicon TCTTGCTGCCCTTTCTGAGGGAAGGACTGT esculentum ACAGTCCTTCCCTCAGAAAGGGCAGCAAGAAGGAGAATACGATT 30

Gly97Ala GGAAAGGGATTTCGAAGCGGGTAATTTAACAGTACCAGATATATC

GGT-GCT TTTGATGGGTNCTAGCACAATCTCATGGGGTT ® ATTACCCGCTTCGAAAT

ATTTCGAAGCGGGTAAT

Glyphosate Resistance [ATTGTTTCCTTGGCACTGACTGCCCA TTCGKATCAACGGCA 33

EPSPS TTGGAGGGCTACCTGCTGGCAAGGTTAAGCTGTCTGGTTCCATC

Lolium rigidum AGCAGCCAATACTTGAGTTCCTTGCTGATGGC

Gly107Ala GCCATCAGCAAGGAACTCAAGTATTGGCTGCTGATGGAACCAGA 34

GGT-GCT CAGCTTAACCTTGCCAGCAGGTAGCCCTCCAATGCCGTTGATCG

AACAGGTGGGCAGTCAGTGCCAAGGAAACAAT

GCTACCTGCTGGCAAGG

CCTTGCCAGCAGGTAGC

14112/01 61:47 pm

AMENDED SHEET 3137.000 — [NY}793559.1

28-12-2001 01939797

Table 11

Genome-Altering Oligos Conferring Imidazolinone and Sulfonylurea Herbicide Resistance

Phenotype, Gene, . Plant & Targeted . . | Alteration Altering Oligos

Sulfonylurea AGCGGATTAGCCGATGCGTTGTTAGATAGTGTTCCTCTTGTAGCA | 37

Resistance ATCACAGGACAAGTCICTCGTCGTATGATTGGTACAGATGCGTTT

ALS CAAGAGACTCCGATTGTTGAGGTAACGCGTT

Arabidopsis thaliana | AACGCGTTACCTCAACAATCGGAGTCTCTTGAAACGCATCTGTAC | 38

Pro197Ser CAATCATACGACGAGAGACTTGTCCTGTGATTGCTACAAGAGGAA

CCT-TCT CACTATCTAACAACGCATCGGCTAATCCGCT ® GACAAGTCICTCGTCGT

ACGACGAGAGACTTGTC ulfonylurea AGCGGATIAGCCGATGCG TAGATAGTGTTCCTCTIGTAGCA :

Resistance ATCACAGGACAAGTCCAGCGTCGTATGATTGGTACAGATGCGTTT

ALS CAAGAGACTCCGATTGTTGAGGTAACGCGTT

Arabidopsis thaliana | AACGCGTTACCTCAACAATCGGAGTCTCTTGAAACGCATCTGTAC | 42

Pro197GIn CAATCATACGACGCTGGACTTGTCCTGTGATTGCTACAAGAGGAA

CCT-CAG CACTATCTAACAACGCATCGGCTAATCCGCT

ACAAGTCCAGCGTCGTC

TACGACGCTGGACTTGT ulfonylurea AGCGGATTAGCCGATGCGTTGTTAGATAGTGTICCICTTGTAGCA

Resistance ATCACAGGACAAGTCCAACGTCGTATGATTGGTACAGATGCGTTT

ALS CAAGAGACTCCGATTGTTGAGGTAACGCGTT

Arabidopsis thaliana | AACGCGTTACCTCAACAATCGGAGTCTCTTGAAACGCATCTGTAC | 46 ® Pro197Gin CAATCATACGACGTTGGACTTGTCCTGTGATTGCTACAAGAGGAA

CCT-CAA CACTATCTAACAACGCATCGGCTAATCCGCT

ACAAGTCCAACGTCGTA

TACGACGTIGGACTTGT

Imidazolinone GACCTTACCTGTTGGATGTGATTTGTCCGCACCAAGAACATGTGT | 49

Resistance TGCCGATGATCCCGAACGGTGGCACTTTCAACGATGTCATAACGG

ALS AAGGAGATGGCCGGATTAAATACTGAGAGAT

Arabidopsis thaliana | ATCTCTCAGTATTTAATCCGGCCATCTCCTTCCGTTATGACATCGT | 50

Ser653Asn TGAAAGTGCCACCGTTCGGGATCATCGGCAACACATGTTCTTGGT

AGT-AAC GCGGACAAATCACATCCAACAGGTAAGGTC

GATCCCGAACGGTGGCA

TGCCACCGTTCGGGATC

1112/01 01:47 pm

AMENDED SHEET $137,009 — NYI7E3558.1

Fs 12-2001 01939797

Phenotype, Gene,

Plant & Targeted . : : . Alteration Altering Oligos

Imidazolinone ACCTTACCTGTTGGATGTGATTTGTCCGCACCAAGAACATG ;

Resistance TGCCGATGATCCCGAATGGTGGCACTTTCAACGATGTCATAACGG

ALS AAGGAGATGGCCGGATTAAATACTGAGAGAT

Arabidopsis thaliana | ATCTCTCAGTATTTAATCCGGCCATCTCCTTCCGTTATGACATCGT | 54

Ser653Asn TGAAAGTGCCACCATTCGGGATCATCGGCAACACATGTTCTTGGT

AGT-AAT GCGGACAAATCACATCCAACAGGTAAGGTC

GATCCCGAATGGTGGCA

TGCCACCATTCGGGATC | 56

Suffonylurea TCOGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTICGC | 57 ® Resistance CATCACGGGCCAGGTCTCCCGCCGCATGATCGGCACCGACGCCT

ALS TCCAGGAGACGCCCATAGTCGAGGTCACCCGCT

Oryza sativa AGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGTG

Pro174Ser CCGATCATGCGGCGGGAGACCTGGCCCGTGATGGCGACCATCG

CCC-TCC GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGGA

GCCAGGTCICCCGCCGC

GCGGCGGGAGACCTGGC EN

Sulfonylurea C 5CT GACGCGCTGCTCGACTCCGTCCCGATGGTCGC 61

Resistance ATCACGGGCCAGGTCCAACGCCGCATGATCGGCACCGACGCCTT

ALS CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC

Oryza sativa GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT | 62

Pro171GIn GCCGATCATGCGGCGTTIGGACCTGGCCCGTGATGGCGACCATCG

CCC-CAA GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGG

CCAGGTCCAACGCCGCA

® TGCGGCGITGGACCTGG | 64

Sulfonylurea CGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGCC | 65

Resistance ATCACGGGCCAGGTCCAGCGCCGCATGATCGGCACCGACGCCTT

ALS CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC

Oryza sativa GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT

Pro171GIn GCCGATCATGCGGCGCTGGACCTGGCCCGTGATGGCGACCATCG

CCC-CAG GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGG

CCAGGTCCAGCGCCGCA

TGCGGCGCTGGACCTGG | 8 "25 Imidazolinone GGCCATACTTIGT TGGATATCATCGTCCCGCACCAGGAGCATGTGC

Resistance TGCCTATGATCCCAAATGGGGGCGCATTCAAGGACATGATCCTGG

ALS ATGGTGATGGCAGGACTGTGTATTAATCTAT

Oryza sativa ATAGATTAATACACAGTCCTGCCATCACCATCCAGGATCATGTCCT | 70 le627Asn TGAATGCGCCCCCATTTGGGATCATAGGCAGCACATGCTCCTGGT

ATT-AAT GCGGGACGATGATATCCAACAAGTATGGCC

GATCCCAAATGGGGGCG

11/12/01 vA)

AMENDED SHEET S131.009 — INYIFE35

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted .

Alteration Altering Ofigos

I I uffonylurea TCCGCGGCTCGCCGACGCGCTGCTCGATTCCGTCCCCATGGTCG 73

Resistance CATCACGGGACAGGTGICGCGACGCATGATTGGCACCGACGCCT

ALS TCCAGGAGACGCCCATCGTCGAGGTCACCCGCT

Zea mays AGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCGGT 74

Pro165Ser GCCAATCATGCGTCGCGACACCTGTCCCGTGATGGCGACCATGG

CCG-TCG GGACGGAATCGAGCAGCGCGTCGGCGAGCGCGGA

GACAGGTGICGCGACGC

GCGTCGCGACACCTGTC

® ulfonylurea [CCGCGCTCBCCGACGCBCTGCTCGATICCGTCCCCATGGTCGE 77

Resistance ATCACGGGACAGGTGCAGCGACGCATGATTGGCACCGACGCCTT

ALS CCAGGAGACGCCCATCGTCGAGGTCACCCGCTC

Zea mays GAGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCGG 78

Pro165GIn TGCCAATCATGCGTCGCTGCACCTGTCCCGTGATGGCGACCATG

CCG-CAG GGGACGGAATCGAGCAGCGCGTCGGCGAGCGCGE

ACAGGTGCAGCGACGCA

TGCGTCGCTIGCACCTGT | 80

Imidazolinone GGCCGTACCTCTTGGATATAATCGTCCCACACCAGGAGCATGTGT

Resistance TGCCTATGATCCCTAATGGTGGGGCTTTCAAGGATATGATCCTGG

ALS ATGGTGATGGCAGGACTGTGTACTGATCTAA

Zea mays TTAGATCAGTACACAGTCCTGCCATCACCATCCAGGATCATATCCT | 82

Ser621Asn TGAAAGCCCCACCATTAGGGATCATAGGCAACACATGCTCCTGGT

AGT-AAT GTGGGACGATTATATCCAAGAGGTACGGCC ® GATCCCTAATGGTGGGG

CCCCACCATTAGGGATC | 84

Imidazolinone GGCCGTACCTCTTGGATATAATCGTCCCACACCAGGAGCATGTGT | 85

Resistance TGCCTATGATCCCTAACGGTGGGGCTTTCAAGGATATGATCCTGG

ALS ATGGTGATGGCAGGACTGTGTACTGATCTAA

Zea mays TTAGATCAGTACACAGTCCTGCCATCACCATCCAGGATCATATCCT

Ser621Asn TGAAAGCCCCACCGTTAGGGATCATAGGCAACACATGCTCCTGGT

AGT-AAC GTGGGACGATTATATCCAAGAGGTACGGCC

GATCCCTAACGGTGGGG

CCCCACCGTTAGGGATC

Sulfonylurea TCCGCGLTCGCCGACGCCCTCCTCGACTCCATCCCCATGGTGGC

Resistance CATCACGGGGCAGGTCTCGCGCCGCATGATCGGCACGGACGCCT

ALS TCCAGGAGACGCCCATCGTCGAGGTCACCCGCT

Lolium multiflorum AGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCCATG

Pro167Ser CCGATCATGCGGCGCGAGACCTGCCCCGTGATGGCCACCATGG

CCG-TCG GGATGGAGTCGAGGAGGGCGTCGGCGAGCGCGGA 14/12/01 01:47 pm

AMEN DED SHEET . 3137.009 — [NY]793559.1

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted .

Alteration Altering Oligos

GGCAGGTCICGCGCCGC

: GCGGCGCGAGACCTGCC

Sufonylurea ~~ J CCGCGCTCGCCGACGLCC ICC ICGACTCOAICCCCATGGIGGCE | 03

Resistance ATCACGGGGCAGGTCCAGCGCCGCATGATCGGCACGGACGCCTT

ALS CCAGGAGACGCCCATCGTCGAGGTCACCCBCTC

Lolium muttiflorum GAGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCCGT| 94

Pro167GIn GCCGATCATGCGGCGCTGGACCTGCCCCGTGATGGCCACCATGG

CCG-CAG GGATGGAGTCGAGGAGGGCCTCGGCCAGCGCGG

GCAGGTCCAGCGCCGCA | 95 9 TGCGGCGCTGGACCTAE % [Imidazolinone |CTGGGCCATACTTGITGGATATCATCGTGCGTCACCAGGAGCATG | 97

Resistance TGCTGCCTATGATCCCTAACGGTGGTGCTTTCAAGGACATTATCA

ALS TGGAAGGTGATGGCAGGATTTCGTATTAAAC

Lolium multiflorum GTTTAATACGAAATCCTGCCATCACCTTCCATGATAATGTCCTTGA

Ser623Asn AAGCACCACCGTTAGGGATCATAGGCAGCACATGCTCCTGGTGA

AGC-AAC GGGACGATGATATCCAACAAGTATGGCCCAG

GATCCCTAACGGTGGTG El

CACCACCGTTAGGGATC

Sulfonylurea ~~ | TCCGCGCTCGCCGACGCTCTCCTCGACTCCATCOCCATGGTCGC 01

Resistarice CATCACGGGCCAGGTCTCACGCCGCATGATCGGCACGGACGCGT

ALS TCCAGGAGACGCCCATAGTGGAGGTCACGCGCT

Hordeum vulgare AGCGCGTGACCTCCACTATGGGCGTCTCCTGGAACGCGTCCGTG | 102

PS Pro68Ser CCGATCATGCGGCGTGAGACCTGGCCCGTGATGGCGACCATGG

CCA-TCA GGATGGAGTCGAGGAGAGCGTCGGCGAGCGCGGA

GCCAGGTCTCACGCCGC

GCGGCGTGAGACCTGGC

Sulfonylurea _ CCGCGCTCGCCGACGCTCTCCTCGACTCCATCGCCATGGICGCC | 105

Resistance ATCACGGGCCAGGTCCAACGCCGCATGATCGGCACGGACGCGTT

ALS CCAGGAGACGCCCATAGTGGAGGTCACGCGCTC

Hordeum vulgare GAGCGCGTGACCTCCACTATGGGCGTCTCCTGGAACGCGTCCGT | 106

Pro68GIn GCCGATCATGCGGCGTIGGACCTGGCCCGTGATGGCGACCATGG

CCA-CAA GGATGGAGTCGAGGAGAGCGTCGGCGAGCGCGG

CCAGGTCCAACGCCGCA

TGCGGCGTTGGACCTGG

1112/01 01:47 pm

AME NDED SHEET 13137.009 — (NY}793559.1

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted .

Alteration Altering Oligos

Imidazolinone CCCAGGG ACCTGCTGGATATCATTGICCCGCATCAGGAGC :

Resistance ACGTGCTGCCTATGATCCCAAACGGTGGTGCTTTCAAGGACATGA

ALS TCATGGAGGGTGATGGCAGGACCTCGTACTGA

Hordeum vulgare TCAGTACGAGGTCCTGCCATCACCCTCCATGATCATGTCCTTGAA 110

Ser524Asn AGCACCACCGITTGGGATCATAGGCAGCACGTGCTCCTGATGCG

AGC-AAC GGACAATGATATCCAGCAGGTACGGCCCTGGG

GATCCCAAACGGTGGTG

CACCACCGTITTGGGATC

Sulfonylurea AGIGGTCICGCTGATGCAATGCTCGATAGTATCCCICTCGTGGCG | 113 ® Resistance ATCACTGGTCAAGTCICTCGTCGGATGATCGGTACCGATGCTTTC

ALS CAGGAAACTCCAATTGTTGAGGTAACAAGGT

Gossypium hirsutum | ACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTAC 114

Pro186Ser CGATCATCCGACGAGAGACTTGACCAGTGATCGCCACGAGAGGG

CCT-TCT ATACTATCGAGCATTGCATCAGCGAGACCACT

GTCAAGTCICTCGTCGG

CCGACGAGAGACTTGAC

Sulfonylurea GTGGTCTOGCTGATGCAATGC TCGATAGTATCCCTCTCGTGGCGA 7

Resistance TCACTGGTCAAGTCCAACGTCGGATGATCGGTACCGATGCTTTCC

ALS AGGAAACTCCAATTGTTGAGGTAACAAGGTC

Gossypium hirsutum | GACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTA | 118

Pro186Gin CCGATCATCCGACGITGGACTTGACCAGTGATCGCCACGAGAGG

CCT-CAA GATACTATCGAGCATTGCATCAGCGAGACCAC

TCAAGTCCAACGTCGGA

® TCCGACGTTGGACTTGA ulfonylurea TGGICTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTIGGCGA | 121

Resistance TCACTGGTCAAGTCCAGCGTCGGATGATCGGTACCGATGCTTTCC

ALS AGGAAACTCCAATTGTTGAGGTAACAAGGTC

Gossypium hirsutum | GACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTA | 122

Pro186GIn CCGATCATCCGACGCTGGACTTGACCAGTGATCGCCACGAGAGG

CCT-CAG GATACTATCGAGCATTGCATCAGCGAGACCAC

TCAAGTCCAGCGTCGGA

TCCGACGCTGGACTTGA

Imidazolinone GACCTTACTIGTTGGATGTGATTGTCCCACATCAAGAACATGTCCT | 125

Resistance GCCTATGATCCCCAATGGAGGCGCTTTCAAAGATGTGATCACAGA

ALS GGGTGATGGAAGAACACAATATTGACCTCA

Gossypium hirsutum | TGAGGTCAATATTGTGTTCTTCCATCACCCTCTGTGATCACATCTT | 126

Ser642Asn TGAAAGCGCCTCCATTGGGGATCATAGGCAGGACATGTTCTTGAT

AGT-AAT GTGGGACAATCACATCCAACAAGTAAGGTC

GATCCCCAATGGAGGCG

1142/01 01:47 pm

AMENDED SHEET 13137.009 — (NY]793559.1

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted . .

Alteration Altering Oligos ulfonylurea TCIGGICTTGOTGATGCACT TCTTGACTCAGTCCCICTTGICGCCA | 129

Resistance TTACTGGGCAAGTTTCCCGGCGTATGATTGGTACTGATGCTTTTCA

ALS AGAGACTCCAATTGTTGAGGTAACTCGAT

Amaranthus ATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTACC | 130 retroflexus AATCATACGCCGGGAAACTTGCCCAGTAATGGCGACAAGAGGGA

Pro192Ser CTGAGTCAAGAAGTGCATCAGCAAGACCAGA

CCC-TCC GGCAAGTTICCCGGCGT ® ACGCCGGGAAACTTGCC

Sulfonylurea CIGGICTTGCTGATGCACTICTTGACTCAGTCCCTCTTGTCGCCAT| 133

Resistance TACTGGGCAAGTTCAACGGCGTATGATTGGTACTGATGCTTTTCA

ALS AGAGACTCCAATTGTTGAGGTAACTCGATC

Amaranthus GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC| 134 retroflexus CAATCATACGCCGITGAACTTGCCCAGTAATGGCGACAAGAGGGA

Pro192Gin CTGAGTCAAGAAGTGCATCAGCAAGACCAG

CCC-CAA GCAAGTTCAACGGCGTA

TACGCCGTTGAACTTGC

Suffonylurea | CTGGTCTTGGIGATGCACT TGTTGACTCAGICCCTCTTGTCGCCAT | 137

Resistance TACTGGGCAAGTTCAGCGGCGTATGATTGGTACTGATGCTTTTCA

ALS AGAGACTCCAATTGTTGAGGTAACTCGATC

Amaranthus GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC| 138 retroflexus CAATCATACGCCGCTGAACTTGCCCAGTAATGGCGACAAGAGGG é Pro192Gin ACTGAGTCAAGAAGTGCATCAGCAAGACCAG

CCC-CAG GCAAGTTCAGCGGCGTA

TACGCCGCTGAACTTGC

Imidazolinone GACCGTATCTG ATGTAATCGTACCACATCAGGAGCATGTGC | 141

Resistance TGCCTATGATCCCTAACGGTGCCGCCTTCAAGGACACCATAACAG

ALS AGGGTGATGGAAGAAGGGCTTATTAGTTGGT

Amaranthus ACCAACTAATAAGCCCTTCTTCCATCACCCTCTGTTATGGTGTCCT | 142 retroflexus TGAAGGCGGCACCGTTAGGGATCATAGGCAGCACATGCTCCTGA

Ser652Asn TGTGGTACGATTACATCCAGCAGATACGGTC

AGC-AAC GATCCCTAACGGTGCCG

CGGCACCGITAGGGATC

1412/01 01:47 pm

AMENDED SHEET 13137.009 — (NY}793559.1

28-12-2001 01939797

Phenotype, Gene, scan!

Plant & Targeted . .

Alteration Altering Oligos ulfonylurea AGCGGCCTCGC TGACGCGCTACTGGATAGCGTCCCCATIGTIGC A5

Resistance TATAACAGGTCAAGTGICACGTAGGATGATAGGTACTGATGCTTTT

ALS 1 CAGGAAACTCCTATTGTTGAGGTAACTAGAT

Nicotiana tabacum | ATCTAGTTACCTCAACAATAGGAGTTTCCTGAAAAGCATCAGTACC | 146

Pro194Ser TATCATCCTACGTGACACTTGACCTGTTATAGCAACAATGGGGAC

CCA-TCA GCTATCCAGTAGCGCGTCAGCGAGGCCGCT

GTCAAGTGTCACGTAGG

CCTACGTGACACTTGAC

Sulfonylurea GCGGCCTCGCTGACGCGCTACTGGATAGCGTCCCCATTGTTGC 149 ® Resistance ATAACAGGTCAAGTGCAACGTAGGATGATAGGTACTGATGCTTTT

ALS 1 CAGGAAACTCCTATTGTTGAGGTAACTAGATC

Nicotiana tabacum | GATCTAGTTACCTCAACAATAGGAGTTTCCTGAAAAGCATCAGTAC | 150

Pro194GIn CTATCATCCTACGTIGCACTTGACCTGTTATAGCAACAATGGGGA

CCA-CAA CGCTATCCAGTAGCGCGTCAGCGAGGCCGC

TCAAGTGCAACGTAGGA

TCCTACGTTGCACTTGA midazoinone | GGCCATACTIGT TGGATGTGATTGTACCTCATCAGGAACATGITIT | 153

Resistance ACCTATGATTCCCAATGGCGGAGCTTTCAAAGATGTGATCACAGA

ALS 1 GGGTGACGGGAGAAGTTCCTATTGAGTTTG

Nicotiana tabacum | CAAACTCAATAGGAACTTCTCCCGTCACCCTCTGTGATCACATCTT | 154

Ser650Asn TGAAAGCTCCGCCATTGGGAATCATAGGTAAAACATGTTCCTGAT

AGT-AAT GAGGTACAATCACATCCAACAAGTATGGCC

GATTCCCAATGGCGGAG

® CTCCGCCATTGGGAATC ulfonylurea AGTGGOC TCGCGGACGCCCTACTGGATAGCGTCCCCATIG] 151

Resistance TATAACCGGTCAAGTGTCACGTAGGATGATCGGTACTGATGCTTT

ALS 2 TCAGGAAACTCCGATTGTTGAGGTAACTAGAT

Nicotiana tabacum | ATCTAGTTACCTCAACAATCGGAGTTTCCTGAAAAGCATCAGTACC | 158

Pro191Ser GATCATCCTACGTGACACTTGACCGGTTATAGCAACAATGGGGAC

CCA-TCA GCTATCCAGTAGGGCGTCCGCGAGGCCACT

GTCAAGTGTCACGTAGG

CCTACGTGACACTTGAC uffonylurea GIGGCCTCGCGGACGCCCTACTGGATAGCGTCCCCATTGTTGCT

Resistance ATAACCGGTCAAGTGCAACGTAGGATGATCGGTACTGATGCTTTT : ALS 2 CAGGAAACTCCGATTGTTGAGGTAACTAGATC

Nicotiana tabacum GATCTAGTTACCTCAACAATCGGAGTTTCCTGAAAAGCATCAGTAC | 162

Pro191Gin CGATCATCCTACGTTGCACTTGACCGGTTATAGCAACAATGGGGA

CCA-CAA CGCTATCCAGTAGGGCGTCCGCGAGGCCAC

TCAAGTGCAACGTAGGA

11/12/01 01:47 pm

AMENDED SHEET 137.009 — PTE38581

28-12-2001 01939797 -h5.

Phenotype, Gene, scan

Plant & Targeted , .

Alteration Altering Oligos

I LC BC midazolinone CATACTTGTTGGATGTGATTGTACCTCATCAGGAACATGTICT 2

Resistance ACCTATGATTCCCAATGGCGGGGCTTTCAAAGATGTGATCACAGA

ALS 2 GGGTGACGGGAGAAGTTCCTATTGACTTTG

Nicotiana tabacum CAAAGTCAATAGGAACTTCTCCCGTCACCCTCTGTGATCACATCTT | 166

Ser647Asn TGAAAGCCCCGCCATTGGGAATCATAGGTAGAACATGTTCCTGAT

AGT-AAT GAGGTACAATCACATCCAACAAGTATGGCC

GATTCCCAATGGCGGGG

CCCCGCCATTGGGAATC

@ Sulfonylurea AGTGGTCTTGCTGATGCTTTATTAGACAGIGITCCAATGGTTGGTA

Resistance TTACTGGTCAAGTTICCAGGAGAATGATTGGAACAGATGCGTTTC

ALS AAGAAACCCCTATTGTTGAGGTAACACGTT

Xanthium spp. AACGTGTTACCTCAACAATAGGGGTTTCTTGAAACGCATCTGTTCC | 170

Pro175Ser AATCATTCTCCTGGAAACTTGACCAGTAATAGCAACCATTGGAACA ccc-TcC CTGTCTAATAAAGCATCAGCAAGACCACT

GTCAAGTTTCCAGGAGA

TCTCCTGGAAACTTGAC

Suffonylurea GTGGTCTTGCTGATGCTTTATTAGACAGTGTTCCAATGGTTGCTAT

Resistance TACTGGTCAAGTTCAAAGGAGAATGATTGGAACAGATGCGTTTCA

ALS AGAAACCCCTATTGTTGAGGTAACACGTTC

Xanthium spp. GAACGTGTTACCTCAACAATAGGGGTTTCTTGAAACGCATCTGTTC| 174

Pro175Gin : CAATCATTCTCCTITGAACTTGACCAGTAATAGCAACCATTGGAAC

CCC-CAA ACTGTCTAATAAAGCATCAGCAAGACCAC ® TCAAGTTCAAAGGAGAA

TTCTCCTITGAACTTGA

Sulfonylurea GIGGTCTTGCTGATGCTTTATTAGACAGTGTTCCAATGGTTGCTAT 77

Resistance TACTGGTCAAGTTCAGAGGAGAATGATTGGAACAGATGCGTTTCA

ALS AGAAACCCCTATTGTTGAGGTAACACGTTC

Xanthium spp. GAACGTGTTACCTCAACAATAGGGGTTTCTTGAAACGCATCTGTTC| 178

Pro175GIn CAATCATTCTCCTCTGAACTTGACCAGTAATAGCAACCATTGGAAC

CCC-CAG ACTGTCTAATAAAGCATCAGCAAGACCAC

TCAAGTTCAGAGGAGAA

: TTCTCCTCTGAACTTGA imidazolinone GGGCCTTACTIGTTGGATGTGATCGTGCCCCATCAAGAACATGIG | 18 : Resistance TTGCCCATGATCCCGAATGGTGGAGGTTTCATGGATGTGATCACC

ALS GAAGGCGACGGCAGAATGAAATATTGAGCTT

Xanthium spp. AAGCTCAATATTTCATTCTGCCGTCGCCTTCGGTGATCACATCCAT | 182

Alab31Asn GAAACCTCCACCATTCGGGATCATGGGCAACACATGTTCTTGATG

GCT-AAT GGGCACGATCACATCCAACAAGTAAGGCCC 1112/01 01:47 pm

AM EN DED S HEET 03137.009 — [NY)793559.1

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted . .

Alteration Altering Oligos

RRR rp——

TGATCCCGAATGGTGGA

TCCACCATTCGGGATCA

Suffonylurea ~ [TCCGGGITIGCTGATGCT CGATTCCGTTCCACTGGTGGC 18

Resistance ATCACGGGGCAGGTGICGCGGCGAATGATTGGGACGGATGCTTT

ALS TCAGGAGACTCCTATTGTTGAGGTAACACGGT

Bassia scoparia ACCGTGTTACCTCAACAATAGGAGTCTCCTGAAAAGCATCCGTCC 186

Pro189Ser CAATCATTCGCCGCGACACCTGCCCCGTGATCGCCACCAGTGGA

CCG-TCG ACGGAATCGAGCAAAGCATCAGCAAACCCGGA

GGCAGGTGICGCGGCGA

® TCGCCGCGACACCTGRC

Sulfonylurea CGGGTTTGCTGATGCITTGCTCGATICGCGTTCCACTGGTGGCGA 7]

Resistance TCACGGGGCAGGTGCAGCGGCGAATGATTGGGACGGATGCTTTT

ALS CAGGAGACTCCTATTGTTGAGGTAACACGGTC

Bassia scoparia GACCGTGTTACCTCAACAATAGGAGTCTCCTGAAAAGCATCCGTC | 190

Pro189GIn CCAATCATTCGCCGCTIGCACCTGCCCCGTGATCGCCACCAGTGG

CCG-CAG AACGGAATCGAGCAAAGCATCAGCAAACCCGG

GCAGGTGCAGCGGCGAA

TTCGCCGCTGCACCTGC

Imidazolinone GACCTTACCTGCTTGATGTGATTGTACCTCATCAGGAGCAIGTGC | 193

Resistance TGCCTATGATTCCTAATGGTGCAGCCTTCAAGGATATCATTAACGA

ALS AGGTGATGGAAGAACAAGTTATTGATGTTC

Bassia scoparia GAACATCAATAACTTGTTCTTCCATCACCTTCGTTAATGATATCCTT | 194

Ser649Asn GAAGGCTGCACCATTAGGAATCATAGGCAGCACATGCTCCTGATG ® AGT-AAT AGGTACAATCACATCAAGCAGGTAAGGTC

GATTCCTAATGGTGCAG

CTGCACCATTAGGAATC ulfonylurea AG AGCAGACGCGATGCTTGACAGIGITCCICTIGTCGE 97

Resistance ATTACAGGACAGGTCTCTCGCCGGATGATCGGTACTGACGCCTTC

ALS 1 CAAGAGACACCAATCGTTGAGGTAACGAGGT

Brassica napus ACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTAC | 198

Pro182Ser CGATCATCCGGCGAGAGACCTGTCCTGTAATGGCGACAAGAGGA

CCT-TCT ACACTGTCAAGCATCGCGTCTGCTAACCCGCT : GACAGGTCICTCGCCGG

CCGGCGAGAGACCTGTC

1/12/01 01:47 pm

AMENDED SHEET 317.008 — (NVI7S35a 1

28-12-2001 01939797 —

Phenotype, Gene,

Plant & Targeted . .

Alteration Altering Oligos

Sulfonylurea [GCGGGTTAGCAGACGCGATGCTTGACAGIGTTCCTCITGTCGCCAT 207

Resistance TTACAGGACAGGTCCAACGCCGGATGATCGGTACTGACGCCTTC : ALS 1 CAAGAGACACCAATCGTTGAGGTAACGAGGTC

Brassica napus GACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTA | 202

Pro182Gin CCGATCATCCGGCGITGGACCTGTCCTGTAATGGCGACAAGAGE

CCT-CAA AACACTGTCAAGCATCGCGTCTGCTAACCCGC

ACAGGTCCAACGCCGGA

Es CC LR

Sulfonylurea GCGGGTTAGCAGACGCGATG ACAGTGTTCCTCITGICGGCA| 205 ® Resistance TTACAGGACAGGTCCAGCGCCGGATGATCGGTACTGACGCCTTC

ALS 1 CAAGAGACACCAATCGTTGAGGTAACGAGGTC

Brassica napus GACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTA | 206

Pro182Gin CCGATCATCCGGCGCTGGACCTGTCCTGTAATGGCGACAAGAGG

CCT-CAG AACACTGTCAAGCATCGCGTCTGCTAACCCGC

ACAGGTCCAGCGCCGGA

TOCGRBREIRGRSTST

Imidazolinone | GACCATACCTGTTGGATGIGATATGICCGCACCAAGAACATGIGT 200]

Resistance TACCGATGATCCCAAATGGTGGCACTTTCAAAGATGTAATAACAG

ALS 1 AAGGGGATGGTCGCACTAAGTACTGAGAGAT

Brassica napus ATCTCTCAGTACTTAGTGCGACCATCCCCTTCTGTTATTACATCTTT| 210

Ser638Asn GAAAGTGCCACCATTTGGGATCATCGGTAACACATGTTCTTGGTG

AGT-AAT CGGACATATCACATCCAACAGGTATGGTC

GATCCCAAATGGTGGCA

® TGCCACCATTTGGGATC

Resistance CATTACAGGACAGGTTCCTCGCCGGATGATCGGTACTGACGCCTT

ALS 2 CCAAGAGACACCAATCGTTGAGGTAACGAGG

Brassica napus CCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTACC | 214

Pro126Ser GATCATCCGGCGAGGAACCTGTCCTGTAATGGCGACAAGAGGAA

CCC-TCC CACTGTCAAGCATCGCGTCTGCTAACCCGCTG

EE NO

CGGCGAGGAACCTGTCC

Suffonylurea AG TTAGCAGACGCGATGCTTGACAGIGTTCCICTIGICGGG I 217

Resistance ATTACAGGACAGGTCACTCGCCGGATGATCGGTACTGACGCCTTC

ALS 2 CAAGAGACACCAATCGTTGAGGTAACGAGGT

Brassica napus ACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTAC | 218

Pro126GIn CGATCATCCGGCGAGTGACCTGTCCTGTAATGGCGACAAGAGGA

CCC-CAG ACACTGTCAAGCATCGCGTCTGCTAACCCGCT

GACAGGTCACTCGCCGG

1112/01 01.47 pm

AMENDED SHEET THT 008 — NI7Saess.

2-2001 01Ys59/9/

Phenotype, Gene,

Plant & Targeted . . SEQID

Eid Altering Oligos NO: midazolinone GACCATACCTGTTGGA ATATGICCGCACCAAGAACATGIGT | 2

Resistance TACCGATGATCCCAAATGGTGGCACTTTCAAAGATGTAATAACAG

ALS 2 AAGGGGATGGTCGCACTAAGTACTGAGAGAT

Brassica napus ATCTCTCAGTACTTAGTGCGACCATCCCCTTCTGTTATTACATCTTT| 222

Ser582Asn GAAAGTGCCACCATTTGGGATCATCGGTAACACATGTTCTTGGTG

AGT-AAT CGGACATATCACATCCAACAGGTATGGTC

GATCCCAAATGGTGGCA

TGCCACCATTTGGGATC

» Sulfonylurea AGCGGGTIAGCCGACGCGATGCTTGACAGTGTICCICTCGTCGC | 225

Resistance CATCACAGGACAGGTCTCTCGCCGGATGATCGGTACTGACGCGT

ALS 3 TCCAAGAGACGCCAATCGTTGAGGTAACGAGGT

Brassica napus ACCTCGTTACCTCAACGATTGGCGTCTCTTGGAACGCGTCAGTAC 226 : Pro179Ser CGATCATCCGGCGAGAGACCTGTCCTGTGATGGCGACGAGAGGA

CCT-TCT ACACTGTCAAGCATCGCGTCGGCTAACCCGCT

GACAGGTCTCTCGCCGG

CCGGCGAGAGACCTGTC

Sulfonylurea GCGGGTTAGCCGACGCGATGCTTGACAGTGTTCCTCTCGTCGCC 0

Resistance ATCACAGGACAGGTCCAACGCCGGATGATCGGTACTGACGCGTT

ALS 3 CCAAGAGACGCCAATCGTTGAGGTAACGAGGTC

Brassica napus GACCTCGTTACCTCAACGATTGGCGTCTCTTGGAACGCGTCAGTA | 230

Pro179GIn CCGATCATCCGGCGITGGACCTGTCCTGTGATGGCGACGAGAGG

CCT-CAA AACACTGTCAAGCATCGCGTCGGCTAACCCGC ® ACAGGTCCAACGCCGGA

TCCGGCGITGGACCTGT uffonylurea G AGCCGACGCGATGCTTGACAGTGTTCCTCTCGTCG 733

Resistance ATCACAGGACAGGTCCAGCGCCGGATGATCGGTACTGACGCGTT

ALS 3 CCAAGAGACGCCAATCGTTGAGGTAACGAGGTC

Brassica napus GACCTCGTTACCTCAACGATTGGCGTCTCTTGGAACGCGTCAGTA | 234

Pro179Gin CCGATCATCCGGCGCTGGACCTGTCCTGTGATGGCGACGAGAGG

CCT-CAG AACACTGTCAAGCATCGCGTCGGCTAACCCGC

ACAGGTCCAGCGCCGGA

TCCGGCGCTGGACCTGT

I RN SR

Imidazofinone GACCGTACCTGTTGGATGTCATCTGTCCGCACCAAGAACATGTGT 37

Resistance TACCGATGATCCCAAATGGTGGCACTTTCAAAGATGTAATAACCG

ALS 3 AAGGGGATGGTCGCACTAAGTACTGAGAGAT

Brassica napus ATCTCTCAGTACTTAGTGCGACCATCCCCTTCGGTTATTACATCTT | 238

Ser635Asn TGAAAGTGCCACCATTTGGGATCATCGGTAACACATGTTCTTGGT

AGT-AAT GCGGACAGATGACATCCAACAGGTACGGTC . 112/01 01:47 pm

AMENDED SHEET 03137.009 ~ [NY]793559.1

28-12-2001 01939797 : Phenotype, Gene, : scan

Plant & Targeted : :

Alteration Altering Oligos

GATCCCAAATGGTGGCA

: TGCCACCATTTGGGATC

Sulfonylurea TCCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGE | 241

Resistance CATCACGGGCCAGGTCTCCCGCCGCATGATCGGCACCGACGCCT

ALS TCCAGGAGACGCCCATAGTCGAGGTCACCCGCT

Oryza sativa AGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGTG | 242

Pro171Ser CCGATCATGCGGCGGGAGACCTGGCCCGTGATGGCGACCATCG

CCC-TCC GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGGA

GCCAGGTCTCCCGCCGC

® GCGGCGGGAGACCTGGC

Sulfonylurea CCGCGCTCGCCGACGLGCTGCTCBACTCCGICCCGATGGICGLC | 245

Resistance ATCACGGGCCAGGTCCAACGCCGCATGATCGGCACCGACGCCTT

ALS CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC

Oryza sativa { GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT | 246

Pro171Gin GCCGATCATGCGGCGITGGACCTGGCCCGTGATGGCGACCATCG

CCC-CAA GGACGGAGTCGAGCAGCGCGBTCGGCGAGCGCGG

CCAGGTCCAACGCCGCA

TGCGGCGTIGGACCTGG ulfonylurea CGCTCGCCGACG GCICGACTCCGTCCCGATGGICGCC | 249 ~~" | Resistance ATCACGGGCCAGGTCCAGCGCCGCATGATCGGCACCGACGCCTT |ALS CCAGGAGACGCCCATAGTCGAGGTCACCCGLTC

Oryza sativa GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT | 250

Pro171GIn GCCGATCATGCGGCGCTGGACCTGGCCCGTGATGGCGACCATCS ® CCC-CAG GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGEG

CCAGGTCCAGCGCCGCA

TGCGGCGCTGGACCTGG

Imidazolinone CATACTTGTTGGATATCATCGICCCGCACCAGGAGCATGTG 25

Resistance TGCCTATGATCCCAAATGGGGGCGCATTCAAGGACATGATCCTGG

ALS ATGGTGATGGCAGGACTGTGTATTAATCTAT

Oryza sativa ATAGATTAATACACAGTCCTGCCATCACCATCCAGGATCATGTCCT | 254

Ser627Asn TGAATGCGCCCCCATTTGGGATCATAGGCAGCACATGCTCCTGGT

AGT-AAT GCGGGACGATGATATCCAACAAGTATGGCC

GATCCCAAATGGGGGCG

CGCCCCCATTTGGGATC

: 11/42/01 01.47 pm

AMENDED SHEET J3437.009 — NYIP3559

28-12-2001 01939797

Phenotype, Gene, : scan

Plant & Targeted . .

Alteration Altering Oligos

Sulfonylurea — GCTCGCAGACGCGTTGCTCGACTCCGICCCCATGGTC 257

Resistance CATCACGGGACAGGTGICGCGACGCATGATTGGCACCGACGCCT : ALS TTCAGGAGACGCCCATCGTCGAGGTCACCCGCT

Zea mays AGCGGGTGACCTCGACGATGGGCGTCTCCTGAAAGGCGTCGGTG | 258

Pro165Ser CCAATCATGCGTCGCGACACCTGTCCCGTGATGGCGACCATGGG

CCG-TCG GACGGAGTCGAGCAACGCGTCTGCGAGCGCAGA

GACAGGTGTCGCGACGC

GCGTCGCGACACCTGTC

Sulfonylurea GCGC AGACGCGTTGCTCGACTCCGICCCCATGGICGCE | 261 ® Resistance ATCACGGGACAGGTGCAGCGACGCATGATTGGCACCGACGCCTT

ALS TCAGGAGACGCCCATCGTCGAGGTCACCCGCTC

Zea mays GAGCGGGTGACCTCGACGATGGGCGTCTCCTGAAAGGCGTCGGT | 262

Pro165GIn GCCAATCATGCGTCGCIGCACCTGTCCCGTGATGGCGACCATGG

CCG-CAG GGACGGAGTCGAGCAACGCGTCTGCGAGCGCAG

ACAGGTGCAGCGACGCA

TGCGTCGCTGCACCTGT

Imdazolinone CCGTACCTCTTGGATATAATCGICCCGCACCAGGAGCATGTGT | 266

Resistance TGCCTATGATCCCTAATGGTGGGGCTTTCAAGGATATGATCCTGG

ALS ATGGTGATGGCAGGACTGTGTATTGATCCGT

Zea mays ACGGATCAATACACAGTCCTGCCATCACCATCCAGGATCATATCC | 266

Ser621Asn TTGAAAGCCCCACCATTAGGGATCATAGGCAACACATGCTCCTGG

AGT-AAT TGCGGGACGATTATATCCAAGAGGTACGGCC

GATCCCTAATGGTGGGG

PS CCCCACCATTAGGGATC

Sulfonylurea AGTGGTCTCGCTGATGCAATGCTCGATAGTATCCCICTCGTGGE 269

Resistance ATCACTGGTCAAGTCTCTCGTCGGATGATCGGTACCGATGCTTTC

ALS CAGGAAACTCCAATTGTTGAGGTAACAAGGT

Gossypium hirsutum | ACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTAC | 270

Pro186Ser CGATCATCCGACGAGAGACTTGACCAGTGATCGCCACGAGAGGG

CCT-TCT ATACTATCGAGCATTGCATCAGCGAGACCACT

GTCAAGTCTCICGTCGG :

CCGACGAGAGACTTGAC

. 25 Sulfonylurea GTGGICTCGCTGATGCAATGCTCGATAGTIATCCCTICTCGIGGLGA 73

Resistance TCACTGGTCAAGTCCAACGTCGGATGATCGGTACCGATGCTTTCC

ALS AGGAAACTCCAATTGTTGAGGTAACAAGGTC

Gossypium hirsutum | GACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTA | 274

Pro186Gin CCGATCATCCGACGITGGACTTGACCAGTGATCGCCACGAGAGGE

CCT-CAA GATACTATCGAGCATTGCATCAGCGAGACCAC

TCAAGTCCAACGTCGGA

11/12/01 01:47 pm

AM EN DED SHEET 03137.009 — [NY]753559.1

28-12-2001 01939797

Phenotype, Gene, : scan

Plant & Targeted .

Alteration Altering Oligos _ ——— — —— ——— — — — ——— ———— — —— = —

CT] TCCGACGTTGGACTTGA ulfonylurea TGGTCTCGCTGATGCAATGCTCGATAGTATCCCICTCGTGGCGA [| 277

Resistance TCACTGGTCAAGTCCAGCGTCGGATGATCGGTACCGATGCTTTCC

ALS AGGAAACTCCAATTGTTGAGGTAACAAGGTC

Gossypium hirsutum | GACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTA 278

Pro186GIn CCGATCATCCGACGCTGGACTTGACCAGTGATCGCCACGAGAGG

CCT-CAG GATACTATCGAGCATTGCATCAGCGAGACCAC

TCAAGTCCAGCGTCGGA

TCCGACGCTIGGACTTGA

® Imidazolinone GACCTTACTTGTTGGATGTGATTGTCCCACATCAAGAACAIGICCT | 281

Resistance GCCTATGATCCCCAATGGAGGGGCTTTCAAAGATGTGATCACAGA

ALS GGGTGATGGAAGAACACAATATTGACCTCA

Gossypium hirsutum | TGAGGTCAATATTGTGTTCTTCCATCACCCTCTGTGATCACATCTT | 282

Ser642Asn TGAAAGCCCCTCCATTGGGGATCATAGGCAGGACATGTTCTTGAT

AGT-AAT GTGGGACAATCACATCCAACAAGTAAGGTC

GATCCCCAATGGAGGGG

CCCCTCCATTGGGGATC

[ Sulfonylurea TCTGGTCTTGCTGATGCACTTCTTGACTCAGICCCICTTGICGCCA| 285

Resistance TTACTGGGCAAGTTICCCGGCGTATGATTGGTACTGATGCTTTTCA

ALS AGAGACTCCAATTGTTGAGGTAACTCGAT

Amaranthus powelli | ATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTACC | 286

Pro192Ser AATCATACGCCGGGAAACTTGCCCAGTAATGGCGACAAGAGGGA

CCC-TCC CTGAGTCAAGAAGTGCATCAGCAAGACCAGA ° I NA

ACGCCGGGAAACTTGCC

Sulfonylurea CTGGTCIT ATGCACTTCTTGACTCAGTCCCTCTTGTGCGGGAT | 289

Resistance TACTGGGCAAGTTCAACGGCGTATGATTGGTACTGATGCTTTTCA

ALS AGAGACTCCAATTGTTGAGGTAACTCGATC

Amaranthus powelli | GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC | 290

Pro192GIn CAATCATACGCCGTTGAACTTGCCCAGTAATGGCGACAAGAGGGA

CCC-CAA CTGAGTCAAGAAGTGCATCAGCAAGACCAG

GCAAGTTCAACGGCGTA

: TACGCCGITGAACTTGC : uffonylurea GTCTTGCTGATGCACTTCTTGACTCAGTCCCICTIGTCGGCAT | 203 : Resistance TACTGGGCAAGTTCAGCGGCGTATGATTGGTACTGATGCTTTTCA

ALS AGAGACTCCAATTGTTGAGGTAACTCGATC

Amaranthus powelli | GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC | 294

Pro192Gin CAATCATACGCCGCTGAACTTGCCCAGTAATGGCGACAAGAGGS

CCC-CA ACTGAGTCAAGAAGTGCATCAGCAAGACCAG 111201 01:47pm

AM E N DED S H EET J3137.009 — [NY]793559.1

28-12-2001 01939797 : Phenotype, Gene,

Plant & Targeted . : Alteration Altering Oligos

GCAAGTTCAGCGGCGTA

TACGCCGCTGAACTTGC midazolinone GACCGTATCTGCTGGATGTAATCGTACCACATCAGGAGCATGIGC | 297

Resistance TGCCTATGATCCCTAACGGTGCCGCCTTCAAGGACACCATAACAG

ALS AGGGTGATGGAAGAAGGGCTTATTAGTTGGT

Amaranthus powelli | ACCAACTAATAAGCCCTTCTTCCATCACCCTCTGTTATGGTGTCCT | 298

Ser652Asn TGAAGGCGGCACCGITAGGGATCATAGGCAGCACATGCTCCTGA

AGC-AAC TGTGGTACGATTACATCCAGCAGATACGGTC

GATCCCTAACGGTGCCG

® CGGCACCGITAGGGATC 14/12/01 01:47 pm

AMENDED SHEET 1ate7.000 — BreaEEat

28-12-2001 01939797

Table 12

Genome-Altering Oligos Conferring Porphyric Herbicide Resistance

Phenotype, Gene, cao

Plant & Targeted . .

Alteration Altering Oligos

Porphyric Herbicide | TCTTGCGCCCTCTTTCTGAATCTGCTGCAAATGCACTCTCAAAACT | 301

Resistant ATATTACCCACCAATGGCAGCAGTATCTATCTCGTACCCGAAAGA

PPO AGCAATCCGAACAGAATGTTTGATAGATGG

Arabidopsis thaliana | CCATCTATCAAACATTCTGTTCGGATTGCTTCTTTCGGGTACGAGA | 302

Val365Met TAGATACTGCTGCCATTGGTGGGTAATATAGTTTTGAGAGTGCATT

GTT-ATG TGCAGCAGATTCAGAAAGAGGGCGCAAGA ® CCCACCAATGGCAGCAG

CTGCTGCCATTGGTGGG

Porphyric Herbicide | TATTACGTCCITCTTTCGGTTGCCGCAGCAGATGCACTTTCAAATT 305

Resistant CTACTATCCCCCAATGGGAGCAGTCACAATTTCATATCCTCAAGAA

PPO GCTATTCGTGATGAGCGTCTGGTTGATGS

Nicotiana tabacum CCATCAACCAGACGCTCATCACGAATAGCTTCTTGAGGATATGAA | 306

Val376Met | ATTGTGACTGCTCCCATTGGGGGATAGTAGAAATTTGAAAGTGCA

GTT-ATG TCTGCTGCGGCAACCGAAAGAGGACGTAATA

TCCCCCAATGGGAGCAG

CTGCTCCCATTGGGGGA

Porphyric Herbicide TGTTGCGTCCGCTTTCGTTGGGTGCAGCAGATGCATTGTCAAAAT 09

Resistant TTTATTATCCTCCGATGGCAGCTGTATCAATTTCATATCCAAAAGA

PPO CGCAATTCGTGCTGACCGGCTGATTGATGG

Cichorium intybus CCATCAATCAGCCGGTCAGCACGAATTGCGTCTTTTGGATATGAA | 310

Val383Met ATTGATACAGCTGCCATCGGAGGATAATAAAATTTTGACAATGCAT ® GTT-ATG CTGCTGCACCCAACGAAAGCGGACGCAACA

TCCTCCGATGGCAGCTG

CAGCTGCCATCGGAGGA

' |Porphyric Herbicide | TCCTTCGTCCACTTTCAGATGTCGCCGCAGAATCTCTTTCAAAATT | 313

Resistant TCATTATCCACCAATGGCAGCTGTGTCACTTTCCTATCCTAAAGAA

PPO GCAATTAGATCAGAGTGCTTGATTGACGG

Spinacia oleracea CCGTCAATCAAGCACTCTGATCTAATTGCTTCTTTAGGATAGGAAA | 314

Val390Met GTGACACAGCTGCCATTGGTGGATAATGAAATTTTGAAAGAGATT . GTT-ATG CTGCGGCGACATCTGAAAGTGGACGAAGGA

TCCACCAATGGCAGCTG

CAGCTGCCATTGGTGGA

11/12/01 01:47 pm

AMENDED SHEET 03137.000 — [NY]763559.1

28-12-2001 01939797

Phenotype, Gene, sca

Plant & Targeted pe

Alteration. Altering Oligos : REET OT I ———————————— —— —— — —————

Porphyric Herbicide TTTGCGTCCACTTTCAAGCGATGCTGCAGATGCTCTATCAAGATT | 317

Resistant CTATTATCCACCGATGGCTGCTGTAACTGTTTCGTATCCAAAGGAA : PPO GCAATTAGAAAAGAATGCTTAATTGATGG

Zea mays CCATCAATTAAGCATTCTTTTCTAATTGCTTCCTTTGGATACGAAAC | 318

Val363Met AGTTACAGCAGCCATCGGTGGATAATAGAATCTTGATAGAGCATC

GTT-ATG TGCAGCATCGCTTGAAAGTGGACGCAAAA

Rg I

CAGCAGCCATCGGTGGA

Porphyric Herbicide | TCTTGCGGCCACTTTCAAGTGATGCAGCAGATGCTCTGTCAATATT | 321 ® Resistant CTATTATCCACCAATGGCTGCTGTAACTGTTTCATATCCAAAAGAA

PPO GCAATTAGAAAAGAATGCTTAATTGACGG

Oryza sativa CCGTCAATTAAGCATTCTTTTCTAATTGCTTCTTTTGGATATGAAAC | 322

Val364Met AGTTACAGCAGCCATTGGTGGATAATAGAATATTGACAGAGCATC

GTT-ATG TGCTGCATCACTTGAAAGTGGCCGCAAGA

TCCACCAATGGCTGCTG

CAGCAGCCATTGGTGGA

Porphyric Herbicide CTGGTCAAGGAGCAGGCGCCCGCCGCCGCCGAGR CT 325

Resistant CCTTCGACTACCCGCCGATGGGCBCCGTGACGCTGTCGTACCCG

PPO CTGAGCGCCGTGCGGGAGGAGCGCAAGGCCTCGG

Chlamydomonas CCGAGGCCTTGCGCTCCTCCCGCACGGCGCTCAGCGGGTACGAC| 326 reinhardtii AGCGTCACGGCGCCCATCGGCGGGTAGTCGAAGGAGCCCAGGG

Val389Met CCTCGGCGGCGGCGGGCGCCTGCTCCTTGACCAG ° EE

A I

Table 13

Genome-Altering Oligos Conferring Triazine Resistance oe eR ee Ee ————— cece

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration ee ___}

Triazine Resistant AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT | 329 } D1 Protein TTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCTT

Arabidopsis thaliana | AGCGGCTTGGCCGGTAGTAGGTATTTG

Ser264Thr CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA | 330

AGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAAATCAATCGGCCAAAAT

AACCGTGAGCAGCTACAATGTTGTAAGTTT

1112/01 01:47 pm

AMENDED SHEET 03137.009 — [NY]7S3659.

28-12-2001 01939797 ee

Phenotype, Gene,

Plant & Targeted Altering Oligos ) Alteration

LL ~erdon 4 —

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

Triazine Resistant AAACTTATAACATCGTAGCCGCTCATGGTTATITIG GATTGAT 33

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Nicotiana tabacum | TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA| 334

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATGTTATAAGTTT

ATATGCTACTTTCAACA

® TGTTGAAAGTAGCATAT

Triazine Resistant AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT 37

D1 Protein CTTCCAATATGCTACTTTTAACAACTCTCGCTCTTTACATTTCTTCT

Populus deltoides TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAAGAAGAAATGTAAAGAG | 338

AGT-ACT CGAGAGTTGTTAAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATATTATAAGTTT

ATATGCTACTTTTAACA

TGTTAAAAGTAGCATAT

Triazine Resistant AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT | 341

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Petunia x hybrida TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA | 342

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATATTATAAGTTT

® ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

D1 Protein CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCC

Magnolia pyramidata | TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA | 346

AGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCAGCTACGATATTATAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACATTTCTTCC : Medicago sativa TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA | 350

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCTGCTACAATATTATAGGTTT

ATATGCTACTTTCAACA

11/12/01 01:47 pm

AM ENDED SHEET 03137.009 — [NY]793553,1

28-12-2001 01939797 —_—

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration . =5 SS EE __| TGTTGAMGTAGCATAT nazine Resistant AAACCTATAATATTGTAGCTGCTCATGGTIATITTGGCCGATTGAT 353 : D1 Protein CTTCCAATATGCAACTTTCAACAATTCTCGTTCTTTACATTTCTTCT

Glycine max TAGCTGCTTGGCCTGTAGTAGGTATTTG

Ser264Thr CAAATACCTACTACAGGCCAAGCAGCTAAGAAGAAATGTAAAGAA | 354

AGT-ACT CGAGAATTGTTGAAAGTTGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCAGCTACAATATTATAGGTTT

ATATGCAACTTTCAACA

TGTTGAAAGTTGCATAT

PY Triazine Resistant | AMACTIACAACATTGTAGCTGGTCACGOT TAT TTGOCCoATI Cn 357

D1 Protein CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCT

Brassica napus TAGCGGCTTGGCCGGTAGTAGGTATTTG

Gly264Thr CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA | 358

GGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

Triazine Resistant | AAMACTTATAATATTGTGGCCGCTCATGGTTATTTTGGCCGATTAAT | 361

D1 Protein CTTCCAATATGCTACTTTTAACAACTCTCGTTCTTTACACTTCTTCT

Oryza sativa TGGCTGCTTGGCCTGTAGTAGGGATTTG

Ser264Thr CAAATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA | 362

AGT-ACT CGAGAGTTGTTAAAAGTAGCATATTGGAAGATTAATCGGCCAAAAT

AACCATGAGCGGCCACAATATTATAAGTTT

ATATGCTACTTTTAACA

Py TGTTAAAAGTAGCATAT

Triazine Resistant AGACTTATAATATTGTGGCTGCTCACGGTIATTTTGGTCGATTAAT 1 365

D1 Protein CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACACTTCTTCT

Zea mays TGGCTGCTTGGCCTGTAGTAGGGATCTG

Ser264Thr CAGATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA | 366

AGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAGATTAATCGACCAAAAT

AACCGTGAGCAGCCACAATATTATAAGTCT

ATATGCTACTTTCAACA

GTTGAAAGTAGCATAT

Triazine Resistant AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT 369 : D1 Protein TTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCTT

Arabidopsis thaliana | AGCGGCTTGGCCGGTAGTAGGTATTTG

Ser264Thr CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA | 370

AGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAAATCAATCGGCCAAAAT

AACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

372 1112/01 01:47 pm

AMENDED SHEET 03137.008 — [NY]793558.1

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Triazine Resistant | AAACTTATAACATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT | 373

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Nicotiana tabacum | TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA | 374

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATGTTATAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

D1 Protein CTTCCAATATGCTACTTTTAACAACTCTCGCTCTTTACATTTCTTCT ® Populus deltoides | TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAAGAAGAAATGTAAAGAG | 378

AGT-ACT CGAGAGTTGTTAAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATATTATAAGTTT

ATATGCTACTTTTAACA

TGTTAAAAGTAGCATAT

Triazine Resistant | AMCTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT | 381

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Petunia x hybrida | TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA | 382

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATATTATAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

PS

D1 Protein CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCC

Magnolia pyramidata | TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA | 386

AGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCAGCTACGATATTATAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACATTTCTTCC

Medicago sativa TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATAGCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA | 390

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCTGCTACAATATTATAGGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

1112/01 01:47 pm

AMENDED SHEET ’ 03137.009 — [NY]793559.1

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Triazine Resistant AAACCTATAATATTGTAGCTGCTCATGGTTATTITGGCCGATTGA 303

D1 Protein CTTCCAATATGCAACTTTCAACAATTCTCGTTCTTTACATTTCTTCT : Glycine max TAGCTGCTTGGCCTGTAGTAGGTATTTG

Ser264Thr CAAATACCTACTACAGGCCAAGCAGCTAAGAAGAAATGTAAAGAA | 394

AGT-ACT CGAGAATTGTTGAAAGTTGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCAGCTACAATATTATAGGTTT

ATATGCAACTTTCAACA

TGTTGAAAGTTGCATAT

Triazine Resistant ARACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT | 397

D1 Protein CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCT @® Brassica napus TAGCGGCTTGGCCGGTAGTAGGTATTTG

Gly264Thr CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA | 308

GGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT [400

Triazine Resistant ~~ | AAACTTATAATATTGTGGCCGCTCATGGTTATTTTGGCCGATTAAT | 401

D1 Protein CTTCCAATATGCTACTTTTAACAACTCTCGTTCTTTACACTTCTTCT

Oryza sativa TGGCTGCTTGGCCTGTAGTAGGGATTTG

Ser264Thr CAAATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA | 402

AGT-ACT CGAGAGTTGTTAAAAGTAGCATATTGGAAGATTAATCGGCCAAAAT

AACCATGAGCGGCCACAATATTATAAGTTT

ATATGCTACTTTTAACA

TGTTAAAAGTAGCATAT

CC) Triazine Resistant AGACTTATAATATTGTGGCTGCTCACGGTTATTTTGGTCGATTAAT ]

D1 Protein CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACACTTCTTCT

Zea mays TGGCTGCTTGGCCTGTAGTAGGGATCTG

Ser264Thr CAGATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA | 406

AGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAGATTAATCGACCAAAAT

AACCGTGAGCAGCCACAATATTATAAGTCT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

Triazine Resistant ~~ | AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT | 409

D1 Protein TTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCTT

Arabidopsis thaliana | AGCGGCTTGGCCGGTAGTAGGTATTTG

Ser264Thr CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA | 410

AGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAAATCAATCGGCCAAAAT

AACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

11/12/04 01:47 pm

AMENDED SHEET 03137.009 — [NY]793559.1

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration

Triazine Resistant AAACCTACAATATTGTGGCT ACGGTTATTTCGGCCGATTGAT | 413

D1 Protein CTTCCAGTATGCTACTTTCAACAACTCCCGTTCTTTACATTTCTTCT

Picea abies TAGCTGCTTGGCCCGTAGCAGGTATCTG

Ser264Thr CAGATACCTGCTACGGGCCAAGCAGCTAAGAAGAAATGTAAAGAA | 414

AGT-ACT CGGGAGTTGTTGAAAGTAGCATACTGGAAGATCAATCGGCCGAAA

TAACCGTGAGCAGCCACAATATTGTAGGTTT

GTATGCTACTTTCAACA

TGTTGAAAGTAGCATAC

Triazine Resistant AAACCTATAATATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT | 417

D1 Protein CTTCCAATATGCTACTTTCAACAATTCTCGCTCTTTACATTTCTTCC ® Vicia faba TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAG | 418

AGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATATTATAGGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

Triazine Resistant AGACTTATAATATTGTGGCTGCTCATGGTTATTTTGGCCGATTAAT | 421

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACACTTCTTCT

Hordeum vulgare TGGCTGCTTGGCCTGTAGTAGGAATCTG

Ser264Thr CAGATTCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA | 422

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATTAATCGGCCAAAA "| TAACCATGAGCAGCCACAATATTATAAGTCT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

®

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACACTTCTTCT

Triticum aestivum TGGCTGCTTGGCCTGTAGTAGGAATCTG

Ser264Thr CAGATTCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA | 426

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATTAATCGGCCAAAA

TAACCATGAGCAGCCACAATATTATAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT riazine Resistant RAACTTATAATATTGTAGC TGCTCATGGTTATTTITGGCCGATIAATC | 429

D1 Protein TTCCAATATGCAACTTTCAACAATTCTCGTTCTTTACATTTCTTCCT

Vigna unguiculata AGCTGCTTGGCCTGTAGTAGGTATTTG

Ser264Thr CAAATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA | 430 . 25 AGT-ACT CGAGAATTGTTGAAAGTTGCATATTGGAAGATTAATCGGCCAAAAT

AACCATGAGCAGCTACAATATTATAAGTTT

ATATGCAACTTTCAACA

TGTTGAAAGTTGCATAT

14/12/01 01:47 pm

AMENDED SHEET 031a7.008 — II7EA598.

28-12-2001 01939797 _ — ee

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration - _—_— TT TIE, BERR. riazine Resistant AAACCTATAATATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT | 43

D1 Protein CTTCCAATATGCAACTTTCAACAACTCTCGTTCTTTACACTTCTTCT : Lotus japonicus TAGCTGCTTGGCCTGTTGTAGGTATCTG

Ser264Thr CAGATACCTACAACAGGCCAAGCAGCTAAGAAGAAGTGTAAAGAA | 434

AGT-ACT CGAGAGTTGTTGAAAGTTGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATATTATAGGTTT

ATATGCAACTTTCAACA

TGTTGAAAGTTGCATAT riazine Resistant AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT | 43

D1 Protein CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCT ® Sinapis alba TAGCGGCTTGGCCGGTAGTAGGTATTTG

Ser264Thr CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA | 438

AGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT ] | 440

Triazine Resistant ~~ | AAACCTATAATATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT | 441

D1 Protein CTTCCAATATGCTACTTTCAACAATTCTCGCTCTTTACATTTCTTCC

Pisum sativum TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAG | 442

AGT-ACT CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATATTATAGGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

®

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACACTTCTTCT

Spinacia oleracea TAGCTGCTTGGCCTGTAGTAGGTATTTG

Ser264Thr CAAATACCTACTACAGGCCAAGCAGCTAAGAAGAAGTGTAAAGAA | 446

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGACCAAAA

TAACCATGAGCAGCTACGATATTATAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

LL OVOAMIABCATAL | 448

Triazine Resistant | AAACTTATAACATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Nicotiana debneyi | TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA | 450

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATGTTATAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

1 STeBIASAIAY | 452 11112001 0147 pm

AMEN DED SHEET 03137.009 — [NY]793669.1

28-12-2001 01939797 —_—

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Triazine Resistant AAA ATAATATCGTAGCCGCTCATGGTTATTIT GATTGA 4

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Solanum nigrum TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA | 454

AGT-ACT CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATATTATAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT

D1 Protein CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC ® Nicotiana TAGCTGCTTGGCCTGTAGTAGGTATCTG plumbaginifolia CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA

Ser264Thr CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

AGT-ACT TAACCATGAGCGGCTACGATGTTATAAGTTT

ATATGCTACTTTCAACA

TGTTGAAAGTAGCATAT | 460 1112/01 01:47 pm

AM EN DED Ss H EET 13137.009 — [NY]793556.1

28-12-2001 01939797

Example 6

Engineering male- or female-sterile plants

Flower development in distantly related dicot plant species is increasingly better understood : and appears to be regulated by a family of genes which encode regulatory proteins. These genes include, for example, AGAMOUS (AG), APETALA1 (AP1), and APETALA3 (AP3) and PISTILLATA (Pl) in

Arabidopsis thaliana, and DEFICIENS A (DEFA), GLOBOSA (GLO), SQUAMOSA (SQUA), and PLENA (PLE) in Antirrhinum majus. Genetic studies have shown that the DEFA, GLO and AP3 genes are essential for petal and stamen development. Sequence analysis of these genes revealed that the gene products contain a conserved MADS box region, a DNA-binding domain. Using these clones as probes, MADS box

C genes have also been isolated from other species including tomato, tobacco, petunia, Brassica napus, and maize.

Altering the expression of these genes results in altered floral morphology. For example, mutations in AP3 and PJ result in male-sterile flowers because petals develop in place of stamens.

The attached tables disclose exemplary oligonucleotide base sequences which can be used to generate site-specific mutations that confer altered floral structures in plants.

Table 14

Oligonucleotides to produce male-sterile plants

Phenotype, Gene,

Plant & Targeted Altering Oligos ) Alteration

Male-sterile TTGTCCTCTCCACCAAATCTCTTCAACAAAAAGATTAAACAAAGAG | 461

AP3 AGAAGAATATGGCGTGAGGGAAGATCCAGATCAAGAGGATAGAGA

Arabidopsis thaliana |ACCAGACAAACAGACAAGTGACGTATTCAA

Arg3Term TTGAATACGTCACTTGTCTGTTTGTCTGGTTCTCTATCCTCTTGATC | 462

AGA-TGA TGGATCTTCCCTCACGCCATATTCTTCTCTCTTTGTTTAATCTTTIT

GTTGAAGAGATTTGGTGGAGAGGACAA

ATATGGCGTGAGGGAAG

CTTCCCTCACGCCATAT

: Male-sterile TCTCCACCAAATCTCTTCAACAAAAAGATTAAACAAAGAGAGAAGA | 465

AP3 ATATGGCGAGAGGGTAGATCCAGATCAAGAGGATAGAGAACCAGA

Arabidopsis thaliana |CAAACAGACAAGTGACGTATTCAAAGAGAA

Lys5Term TTCTCTTTGAATACGTCACTTGTCTGTTTGTCTGGTTCTCTATCCTC | 466

AAG-TAG TTGATCTGGATCTACCCTCTCGCCATATTCTTCTCTCTTTGTTTAAT

CTTTTTGTTGAAGAGATTTGGTGGAGA

CGAGAGGGTAGATCCAG

14/12001 04:47 pm

AMENDED SH EET 13137.009 — [NY]793559.1

28-12-2001 01939797 [_ CTGGATCTACCCTCICG

Male-sterile CCAAATCTCTTCAACAAAAAGATTAAACAAAGAGAGAAGAATATGG | 469 : AP3 CGAGAGGGAAGATCTAGATCAAGAGGATAGAGAACCAGACAAACA

Arabidopsis thaliana | GACAAGTGACGTATTCAAAGAGAAGGAATG

Gin7Term CATTCCTTCTCTTTGAATACGTCACTTGTCTGTTTGTCTGGTTCTCT | 470

CAG-TAG ATCCTCTTGATCTAGATCTTCCCTCTCGCCATATTCTTCTCTCTTTG

TTTAATCTTTTTGTTGAAGAGATTTGG

GGAAGATCTAGATCAAG

CTTGATCTAGATCTTCC

Male-sterile CTCTTCAACAAAAAGATTAAACAAAGAGAGAAGAATATGGCGAGAG| 473

AP3 GGAAGATCCAGATCTAGAGGATAGAGAACCAGACAAACAGACAAG

Arabidopsis thaliana | TGACGTATTCAAAGAGAAGGAATGGTTTAT ® Lys9Term ATAAACCATTCCTTCTCTTTGAATACGTCACTTGTCTGTTTGTCTGG | 474

AAG-TAG TTCTCTATCCTCTAGATCTGGATCTTCCCTCTCGCCATATTCTTCTC

TCTTTGTTTAATCTTTTTGTTGAAGAG

TCCAGATCTAGAGGATA

TATCCTCTAGATCTGGA

Malesternie |AGAGGGAAGATCCAGAICAAGAGGATAGAGAACCAGACCAACCG | 477

AP3 ACAAGTGACGTATTCTTAGAGAAGAAATGGTTTGTTCAAGAAAGCT

Brassica oleracea CACGAGCTTACAGTTTTATGTGATGCTAGGG

Lys23Term CCCTAGCATCACATAAAACTGTAAGCTCGTGAGCTTTCTTGAACAA | 478

AAG-TAG ACCATTTCTTCTCTAAGAATACGTCACTTGTCGGTTGGTCTGGTTC : TCTATCCTCTTGATCTGGATCTTCCCTCT

CGTATTCTTAGAGAAGA

TCTTCTCTAAGAATACG

® AP3 GTGACGTATTCTAAGTGAAGAAATGGTTTGTTCAAGAAAGCTCACG

Brassica oleracea AGCTTACAGTTTTATGTGATGCTAGGGTTT

Arg24Term AAACCCTAGCATCACATAAAACTGTAAGCTCGTGAGCTTTCTTGAA | 482

AGA-TGA CAAACCATTTCTTCACTTAGAATACGTCACTTGTCGGTTGGTCTGG

TTCTCTATCCTCTTGATCTGGATCTTCCC

ATTCTAAGTIGAAGAAAT

ATTTCTTCACTTAGAAT

Male-sterile AAGATCCAGATCAAGAGGATAGAGAACCAGACCAACCGACAAGTG | 485

AP3 ACGTATTCTAAGAGATGAAATGGTTTGTTCAAGAAAGCTCACGAGC

Brassica oleracea TTACAGTTTTATGTGATGCTAGGGTTTCGA

Arg25Term TCGAAACCCTAGCATCACATAAAACTGTAAGCTCGTGAGCTTTCTT | 486

AGA-TGA GAACAAACCATTTCATCTCTTAGAATACGTCACTTGTCGGTTGGTC : TGGTTCTCTATCCTCTTGATCTGGATCTT

CTAAGAGATGAAATGGT

ACCATTTCATCTCTTAG

1112/01 01:47 pm

AMENDED SHEET 03137.009 — [NY[73559.1

28-12-2001 01939797

Male-sterile TCAAGAGGATAGAGAACCAGACCAACCGACAAGTGACGTATTCTA | 489

AP3 AGAGAAGAAATGGTTAGTTCAAGAAAGCTCACGAGCTTACAGTTTT } Brassica oleracea ATGTGATGCTAGGGTTTCGATTATCATGTT

Leu28Term AACATGATAATCGAAACCCTAGCATCACATAAAACTGTAAGCTCGT

TTG-TAG GAGCTTTCTTGAACTAACCATTTCTTCTCTTAGAATACGTCACTTGT

CGGTTGGTCTGGTTCTCTATCCTCTTGA

AAATGGTTAGTTCAAGA

TCTTGAACTAACCATTT

\/ ale-sterile A AA A A A AA A ATA A AA AAA AA A AQ

AP3 CAGGCAGGTCACCTAGTCCAAGAGAAGAAATGGTTTGTTCAAGAA Bo

Brassica napus AGCACACGAGCTCTCTGTTCTCTGTGATGCT

Tyr21Term AGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTTGAACAAACC | 494 é TAC-TAG ATTTCTTCTCTTGGACTAGGTGACCTGCCTGTTTGTTTGGTTCTCTA

TCCTCTTAATCTGGATCTTCCCTCGAGCC

GTCACCTAGTCCAAGAG

CTCTTGGACTAGGTGAC | 496

Male-sterile CGAGGGAAGATCCAGATTAAGAGGATAGAGAACCAAACAAACAGG | 497

AP3 CAGGTCACCTACTCCTAGAGAAGAAATGGTTTGTTCAAGAAAGCAC

Brassica napus ACGAGCTCTCTGTTCTCTGTGATGCTAAAG

Lys23Term CTTTAGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTTGAACAA | 498

AAG-TAG ACCATTTCTTCTCTAGGAGTAGGTGACCTGCCTGTTTGTTTGGTTC

TCTATCCTCTTAATCTGGATCTTCCCTCG

CCTACTCCTAGAGAAGA | 499

TCTTCTCTAGGAGTAGG

Male-sterile GGGAAGATCCAGATTAAGAGGATAGAGAACCAAACAAACAGGCAG

AP3 GTCACCTACTCCAAGIGAAGAAATGGTTTGTTCAAGAAAGCACACG

Brassica napus AGCTCTCTGTTCTCTGTGATGCTAAAGTTT @ Arg24Term AAACTTTAGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTTGAA | 502

AGA-TGA CAAACCATTTCTTCACTTGGAGTAGGTGACCTGCCTGTTTGTTTGG

TTCTCTATCCTCTTAATCTGGATCTTCCC

ACTCCAAGTGAAGAAAT

ATTTCTTCACTTGGAGT | 504

Male-sterile AAGATCCAGATTAAGAGGATAGAGAACCAAACAAACAGGCAGGTC

AP3 ACCTACTCCAAGAGATGAAATGGTTTGTTCAAGAAAGCACACGAG

Brassica napus CTCTCTGTTCTCTGTGATGCTAAAGTTTCCA

Arg25Term TGGAAACTTTAGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTT © 25 AGA-TGA GAACAAACCATTTCATCTCTTGGAGTAGGTGACCTGCCTGTTTGTT

TGGTTCTCTATCCTCTTAATCTGGATCTT

: CCAAGAGATGAAATGGT

ACCATTTCATCTCTTGG | 508 1142/01 01:47 pm

AMENDED SHEET 03137.009 — [NY)763559.1

28-12-2001 01939797

Viale-s erie A A! AAA AAA AS AR AAAA AA A A A (JU

DEFA TAGTGGTTCGATGGCTIGAGGGAAGATCCAGATTAAGAGGATAGA

Antirrhinum majus GAACCAAACAAACAGGCAGGTCACCTACTCCA

Arg3Term TGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTCTATCCTCTTAAT | 510

CGA-TGA CTGGATCTTCCCTCAAGCCATCGAACCACTACCACTACTGCTCTTG

TTTTCTTCTTCCAGCTTTCCTTTCTCTCC

CGATGGCTTGAGGGAAG

CTTCCCTCAAGCCATCG

Male-sterile AAAGGAAAGCTGGAAGAAGAAAACAAGAGCAGTAGTGGTAGTGGT | 513

DEFA TCGATGGCTCGAGGGTAGATCCAGATTAAGAGGATAGAGAACCAA

Antirrhinum majus ACAAACAGGCAGGTCACCTACTCCAAGAGAA

Lys5Term TTCTCTTGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTCTATCCT | 514 é AAG-TAG CTTAATCTGGATCTACCCTCGAGCCATCGAACCACTACCACTACTG

CTCTTGTTITCTTCTTCCAGCTTTCCTTT

CTCGAGGGTAGATCCAG

CTGGATCTACCCTCGAG

Male-sterile AAGCTGGAAGAAGAAAACAAGAGCAGTAGTGGTAGTGGTTCGATG | 517

DEFA GCTCGAGGGAAGATCTAGATTAAGAGGATAGAGAACCAAACAAAC

Antirrhinum majus AGGCAGGTCACCTACTCCAAGAGAAGAAATG

Gln7Term CATTTCTTCTCTTGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTC | 518

CAG-TAG TATCCTCTTAATCTAGATCTTCCCTCGAGCCATCGAACCACTACCA

CTACTGCTCTTGTTTTCTTCTTCCAGCTT

GGAAGATCTAGATTAAG [519

CTTAATCTAGATCTTCC

Male-sterile GAAGAAGAAAACAAGAGCAGTAGTGGTAGTGGTTCGATGGCTCGA | 521

DEFA GGGAAGATCCAGATTTAGAGGATAGAGAACCAAACAAACAGGCAG

Antirrhinum majus GTCACCTACTCCAAGAGAAGAAATGGTTTGT o Lys9Term ACAAACCATTTCTTCTCTTGGAGTAGGTGACCTGCCTGTTTGTTIG | 522

AAG-TAG GTTCTCTATCCTCTAAATCTGGATCTTCCCTCGAGCCATCGAACCA

CTACCACTACTGCTCTTGTTTTICTTCTTC

TCCAGATTTAGAGGATA

TATCCTCTAAATCTGGA

AP3 TATGGCTCGTGGGTAGATCCAGATCAAGAGAATAGAGAACCAAAC BH

Nicotiana tabacum AAACAGACAAGTCACTTATTCTAAGAGAA

Lys5Term TTCTCTTAGAATAAGTGACTTGTCTGTTTGTTTGGTTCTCTATICTC | 526 © 25 AAG-TAG TTGATCTGGATCTACCCACGAGCCATAGTTTTTTTTTCTTTTTGCTC

AAAGTTTGAGATCTTAAGAATTACTGA

CTCGTGGGTAGATCCAG

CTGGATCTACCCACGAG

1412/01 01:47 pm

AMENDED SHEET 03137.009 — [NY1790559.

28-12-2001 01939797

Male-sterile ATTCTTAAGATCTCAAACTTTGAGCAAAAAGAAAAAAAAACTATGGC| 529

AP3 TCGTGGGAAGATCTAGATCAAGAGAATAGAGAACCAAACAAACAG

Nicotiana tabacum ACAAGTCACTTATTCTAAGAGAAGAAATG

GIn7Term CATTTCTTCTCTTAGAATAAGTGACTTGTCTGTTTGTTTGGTTCTCT | 530

CAG-TAG ATTCTCTTGATCTAGATCTTCCCACGAGCCATAGTTTTTTTTTCTTT

TTGCTCAAAGTTTGAGATCTTAAGAAT

GGAAGATCTAGATCAAG

CTTGATCTAGATCTTCC

Male-sterile AAGATCTCAAACTTTGAGCAAAAAGAAAAAAAAACTATGGCTCGTG | 533

AP3 GGAAGATCCAGATCTAGAGAATAGAGAACCAAACAAACAGACAAG

Nicotiana tabacum TCACTTATTCTAAGAGAAGAAATGGACTTT

Lys9Term AAAGTCCATTTCTTCTCTTAGAATAAGTGACTTGTCTGTTTGTTTGG | 534

AAG-TAG TTCTCTATTCTCTAGATCTGGATCTTCCCACGAGCCATAGTTTTTTT ® TTCTTTTTGCTCAAAGTTTGAGATCTT

TATTCTCTAGATCTGGA

Male-sterie ~~ |ATCTCAAACTTTGAGCAAAAAGAAAAAAAAACTATGGCTCGTGGGA | 537

AP3 AGATCCAGATCAAGTGAATAGAGAACCAAACAAACAGACAAGTCA

Nicotiana tabacum CTTATTCTAAGAGAAGAAATGGACTTTTCA

Arg10Tem TGAAAAGTCCATTTCTTCTCTTAGAATAAGTGACTTGTCTGTTTGTT | 538

AGA-TGA TGGTTCTCTATTCACTTGATCTGGATCTTCCCACGAGCCATAGTTT

TTTTTTCTTTTTGCTCAAAGTTTGAGAT :

AGATCAAGTGAATAGAG

CTCTATTCACTTGATCT | 540

AP3 CAGACAAGTAACTTAGTCAAAACGAAGGGATGGTCTTTTCAAGAAG BB

Medicago sativa GCCAATGAGCTCACTGTTCTTTGTGATGCT

C Tyr21Term AGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAAAAGACCA| 542

TAC-TAG TCCCTTCGTTTTGACTAAGTTACTTGTCTGTTCGTTGTGTTCTCTAT

TCTCTTGATCTGGATCTTTCCTCGAGCC

GTAACTTAGTCAAAACG

CGTTTTGACTAAGTTAC

Male-sterile CTCGAGGAAAGATCCAGATCAAGAGAATAGAGAACACAACGAACA | 545

AP3 GACAAGTAACTTACTGAAAACGAAGGGATGGTCTTTTCAAGAAGG

Medicago sativa CCAATGAGCTCACTGTTCTTTGTGATGCTAA

Ser22Term TTAGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAAAAGAC | 546

TCA-TGA CATCCCTTCGTTTTCAGTAAGTTACTTGTCTGTTCGTTGTGTTICTCT

ATTCTCTTGATCTGGATCTTTCCTCGAG

AACTTACTGAAAACGAA

TTCGTTTTCAGTAAGTT

11201 01:47 pm

AMENDED SHEET 13137.009 — (NY]783559.1

28-12-2001 01939797

Male-sterile CGAGGAAAGATCCAGATCAAGAGAATAGAGAACACAACGAACAGA | 549

AP3 CAAGTAACTTACTCATAACGAAGGGATGGTCTTTTCAAGAAGGCCA ) Medicago sativa ATGAGCTCACTGTTCTTTGTGATGCTAAGG

Lys23Term CCTTAGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAAAAG | 550

AAA-TAA ACCATCCCTTCGTTATGAGTAAGTTACTTGTCTGTTCGTTGTGTTCT : CTATTCTCTTGATCTGGATCTTTCCTCG

CTTACTCATAACGAAGG

CCTTCGTTATGAGTAAG

AP3 GTAACTTACTCAAAATGAAGGGATGGTCTTTTCAAGAAGGCCAATG

Medicago sativa AGCTCACTGTTCTTTGTGATGCTAAGGTTT

Arg24Term AAACCTTAGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAA | 554

CGA-TGA AAGACCATCCCTTCATTTTGAGTAAGTTACTTGTCTGTTCGTTGTGT ® TCTCTATTCTCTTGATCTGGATCTTTCC

ACTCAAAATGAAGGGAT

ATCCCTTCATTTTGAGT

DEF4 AGGCAAGTGACTTAGTCAAAGAGAAGAAATGGGCTATTCAAGAAG

Solanum tuberosum |GCTAATGAACTTACAGTTCTTTGTGATGCT

Tyr21Term AGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAATAGCCCA

TAT-TAG TTTCTTCTCTTTGACTAAGTCACTTGCCTATTTGTTTGGTTTTCTATT

TTCTTGATCTGGATCTTACCACGAGCC

GTGACTTAGTCAAAGAG

CTCTTTGACTAAGTCAC | 560

Male-sterile CTCGTGGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAATAG | 561 : DEF4 GCAAGTGACTTATTGAAAGAGAAGAAATGGGCTATTCAAGAAGGC

Solanum tuberosum | TAATGAACTTACAGTTCTTTGTGATGCTAA

C Ser22Term TTAGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAATAGCC | 562

TCA-TGA CATTTCTTCTCTTTCAATAAGTCACTTGCCTATTTGTTTGGTTTTCTA

TTTTCTTGATCTGGATCTTACCACGAG

GACTTATTGAAAGAGAA

TTCTCTTTCAATAAGTC

Male-sterile CGTGGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAATAGG | 565

DEF4 CAAGTGACTTATTCATAGAGAAGAAATGGGCTATTCAAGAAGGCTA

Solanum tuberosum |ATGAACTTACAGTTCTTTGTGATGCTAAAG

Lys23Term CTTTAGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAATAG

AAG-TAG CCCATTTCTTCTCTATGAATAAGTCACTTGCCTATTTGTTTGGTTTT

CTATTTTCTTGATCTGGATCTTACCACG

CTTATTCATAGAGAAGA

[ICTICTCTATGAATAAG | 568 14/12/01 01:47 pm

AMENDED SHEET 03137.009 — INY}7935531

28-12-2001 01939797

Male-sterile GGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAATAGGCAA | 569

DEF4 GTGACTTATTCAAAGTGAAGAAATGGGCTATTCAAGAAGGCTAATG

Solanum tuberosum [AACTTACAGTTCTTTGTGATGCTAAAGTTT

Arg24Term AAACTTTAGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAAT | 570

AGA-TGA AGCCCATTTCTTCACTTTGAATAAGTCACTTGCCTATTTGTTTGGTT : TTCTATTTTCTTGATCTGGATCTTACC

ATTCAAAGTGAAGAAAT

ATTTCTTCACTTTGAAT

[Male-sfenle AATGAZ A A AAA AA i

AP3 GATTTCTAGTACTTGAAAACTTCATGAGTTTATAAGTCCCTCTATCA

Lycopersicon CGACCAAACAATTGTTCGATCTGTACC esculentum GGTACAGATCGAACAATTGTTTGGTCGTGATAGAGGGACTTATAAA | 574 é Gly27Term CTCATGAAGTTTTCAAGTACTAGAAATCATAACAATTGAAACTTTAG

GGA-TGA CATCACAAAGAACAGTAAGTTCATTAGC

CTAGTACTTIGAAAACTT

AAGTTTTCAAGTACTAG

Male-sterile AATGAACTTACTGTTCTTTGTGATGCTAAAGTTTCAATTGTTATGAT | 577

AP3 TTCTAGTACTGGATAACTTCATGAGTTTATAAGTCCCTCTATCACGA

Lycopersicon CCAAACAATTGTTCGATCTGTACCAGA esculentum TCTGGTACAGATCGAACAATTGTTTGGTCGTGATAGAGGGACTTAT | 578

Lys28Term AAACTCATGAAGTTATCCAGTACTAGAAATCATAACAATTGAAACTT

AAA-TAA TAGCATCACAAAGAACAGTAAGTTCATT

GTACTGGATAACTTCAT

ATGAAGTTATCCAGTAC 580

Male-sterile ACTGTTCTTTGTGATGCTAAAGTTTCAATTGTTATGATTTCTAGTAC | 581

AP3 TGGAAAACTTCATTAGTTTATAAGTCCCTCTATCACGACCAAACAAT

Lycopersicon TGTTCGATCTGTACCAGAAGACTATTG ( esculentum CAATAGTCTTCTGGTACAGATCGAACAATTGTTTGGTCGTGATAGA | 582

Glu31Term GGGACTTATAAACTAATGAAGTTTTCCAGTACTAGAAATCATAACA

GAG-TAG ATTGAAACTTTAGCATCACAAAGAACAGT

AACTTCATTAGTTTATA

TATAAACTAATGAAGTT | 584

Male-sterile ATTGTTATGATTTCTAGTACTGGAAAACTTCATGAGTTTATAAGTCC | 585

AP3 CTCTATCACGACCTAACAATTGTTCGATCTGTACCAGAAGACTATT

Lycopersicon GGAGTTGATATTTGGACTACTCACTATG esculentum CATAGTGAGTAGTCCAAATATCAACTCCAATAGTCTTCTGGTACAG

Lys40Term ATCGAACAATTGTTAGGTCGTGATAGAGGGACTTATAAACTCATGA

AAA-TAA AGTTTTCCAGTACTAGAAATCATAACAAT

TCACGACCTAACAATTG

CAATTGTTAGGTCGTGA

1112/01 01:47 pm

AMENDED SHEET 03137.009 — [NY]793559.1

28-12-2001 | 01939797 1 ale-sterile AAGA A ) AAA R AGAA A A oe

AP3 ACAGGCAGGTGACCTAGTCCAAGCGCCGGTCGGGGATCATGAAG

Triticum aestivum AAGGCGCGGGAGCTCACCGTGCTCTGCGACGCC : Tyr21Term GGCGTCGCAGAGCACGGTGAGCTCCCGCGCCTTCTTCATGATCC

TAC-TAG CCGACCGGCGCTTGGACTAGGTCACCTGCCTGTTGGTGGCGTTCT

CGATCCGCTTTATCTCAATCTTCCCCCGCCCC

GTGACCTAGTCCAAGCG

CGCTTGGACTAGGTCAC

Male-sterile CGGGGGAAGATTGAGATAAAGCGGATCGAGAACGCCACCAACAG | 593

AP3 GCAGGTGACCTACTCCTAGCGCCGGTCGGGGATCATGAAGAAGG

Triticum aestivum CGCGGGAGCTCACCGTGCTCTGCGACGCCCAGG

Lys23Term CCTGGGCGTCGCAGAGCACGGTGAGCTCCCGCGCCTTCTTCATG | 594

AAG-TAG ATCCCCGACCGGCGCTAGGAGTAGGTCACCTGCCTGTTGGTGGC ® GTTCTCGATCCGCTTTATCTCAATCTTCCCCCG

CCTACTCCTAGCGCCGG

CCGGCGCTAGGAGTAGG | 596

Male-sterile TTGAGATAAAGCGGATCGAGAACGCCACCAACAGGCAGGTGACCT | 597

AP3 ACTCCAAGCGCCGGTAGGGGATCATGAAGAAGGCGCGGGAGCTC

Triticum aestivum ACCGTGCTCTGCGACGCCCAGGTCGCCATCAT

Ser26Term ATGATGGCGACCTGGGCGTCGCAGAGCACGGTGAGCTCCCGCGC

TCG-TAG CTTCTTCATGATCCCCTACCGGCGCTTGGAGTAGGTCACCTGCCT

GTTGGTGGCGTTCTCGATCCGCTTTATCTCAA

GCGCCGGTAGGGGATCA

: TGATCCCCTACCGGCGC | 600

Male-sterile CGGATCGAGAACGCCACCAACAGGCAGGTGACCTACTCCAAGCG | 601

AP3 CCGGTCGGGGATCATGTAGAAGGCGCGGGAGCTCACCGTGCTCT

Triticum aestivum GCGACGCCCAGGTCGCCATCATCATGTTCTCCT @® Lys30Term AGGAGAACATGATGATGGCGACCTGGGCGTCGCAGAGCACGGTG | 602

AAG-TAG AGCTCCCGCGCCTTCTACATGATCCCCGACCGGCGCTTGGAGTAG

GTCACCTGCCTGTTGGTGGCGTTCTCGATCCG

GGATCATGTAGAAGGCG

CGCCTTCTACATGATCC

Viale-sterile AAGA AGA AR A AGAA A s ol

Sitky1 ACCGCCAGGTGACCTAGTCCAAGCGCCGGACGGGGATCATGAAG

Zea mays AAGGCACGCGAGCTCACCGTGCTCTGCGACGCC

Tyr21Term GGCGTCGCAGAGCACGGTGAGCTCGCGTGCCTTCTTCATGATCCC © 25 TAC-TAG CGTCCGGCGCTTGGACTAGGTCACCTGGCGGTTGGTGGCGTTCT

CGATCCGCTTGATCTCGATCTTGCCGCGCCCC

GTGACCTAGTCCAAGCG

CGCTTGGACTAGGTCAC

11/12/01 01:47 pm

AMENDED SHEET 03137.009 — [NY]793559.1

28-12-2001 01939797

Male-sterile CGCGGCAAGATCGAGATCAAGCGGATCGAGAACGCCACCAACCG

Sitky1 CCAGGTGACCTACTCCTAGCGCCGGACGGGGATCATGAAGAAGG

Zea mays CACGCGAGCTCACCGTGCTCTGCGACGCCCAGG

Lys23Term CCTGGGCGTCGCAGAGCACGGTGAGCTCGCGTGCCTTCTTCATG | 610

AAG-TAG ATCCCCGTCCGGCGCTAGGAGTAGGTCACCTGGCGGTTGGTGGC

GTTCTCGATCCGCTTGATCTCGATCTTGCCGCG

CCTACTCCTAGCGCCGG

CCGGCGCTAGGAGTAGG

Male-sterile CGGATCGAGAACGCCACCAACCGCCAGGTGACCTACTCCAAGCG | 613

Silky1 CCGGACGGGGATCATGTAGAAGGCACGCGAGCTCACCGTGCTCT

Zea mays GCGACGCCCAGGTCGCCATCATCATGTTCTCCT

Lys30Term AGGAGAACATGATGATGGCGACCTGGGCGTCGCAGAGCACGGTG | 614

AAG-TAG AGCTCGCGTGCCTTCTACATGATCCCCGTCCGGCGCTTGGAGTAG ® GTCACCTGGCGGTTGGTGGCGTTCTCGATCCG

GGATCATGTAGAAGGCA

TGCCTTCTACATGATCC

Male-sterile ATCGAGAACGCCACCAACCGCCAGGTGACCTACTCCAAGCGCCG | 617

Silky GACGGGGATCATGAAGTAGGCACGCGAGCTCACCGTGCTCTGCG

Zea mays ACGCCCAGGTCGCCATCATCATGTTCTCCTCCA

Lys31Term TGGAGGAGAACATGATGATGGCGACCTGGGCGTCGCAGAGCACG | 618

AAG-TAG GTGAGCTCGCGTGCCTACTTCATGATCCCCGTCCGGCGCTTGGAG

TAGGTCACCTGGCGGTTGGTGGCGTTCTCGAT

TCATGAAGTAGGCACGC

GCGTGCCTACTTCATGA

Vale-stenle 4 A AGAGAGATA A :

AP3 GGCCATGGGGAGGGGCTAGATCGAGATCAAGCGGATCGAGAACG

Oryza sativa CGACCAACAGGCAGGTGACCTACTCGAAGCGCC

C Lys5Term GGCGCTTCGAGTAGGTCACCTGCCTGTTGGTCGCGTTCTCGATCC | 622

AAG-TAG GCTTGATCTCGATCTAGCCCCTCCCCATGGCCGCCCCCTGCAGCA

GCTATCTCTCTCGCCGGACAATGCAGCTAGC

GGAGGGGCTAGATCGAG

CTCGATCTAGCCCCTCC

AP3 GGGGAGGGGCAAGATCTAGATCAAGCGGATCGAGAACGCGACCA

Oryza sativa ACAGGCAGGTGACCTACTCGAAGCGCCGCACGG

Glu7 Term CCGTGCGGCGCTTCGAGTAGGTCACCTGCCTGTTGGTCGCGTTCT | 626

GAG-TAG CGATCCGCTTGATCTAGATCTTGCCCCTCCCCATGGCCGCCCCCT

GCAGCAGCTATCTCTCTCGCCGGACAATGCA

: GCAAGATCTAGATCAAG

CTTGATCTAGATCTTGC

111201 01:47 pm

AMENDED SHEET 03137.009 — [NY]793559.1

28-12-2001 01939797

Male-sterile GTCCGGCGAGAGAGATAGCTGCTGCAGGGGGCGGCCATGGGGA | 629

AP3 GGGGCAAGATCGAGATCTAGCGGATCGAGAACGCGACCAACAGG

Oryza sativa CAGGTGACCTACTCGAAGCGCCGCACGGGGATCA

Lys9Term TGATCCCCGTGCGGCGCTTCGAGTAGGTCACCTGCCTGTTGGTCG| 630

AAG-TAG CGTTCTCGATCCGCTAGATCTCGATCTTGCCCCTCCCCATGGCCG : CCCCCTGCAGCAGCTATCTCTCTCGCCGGAC

TCGAGATCTAGCGGATC

GATCCGCTAGATCTCGA

GAGAGATAGCTGCTGCAGGGGGCGGCCATGGGGAGGGGCAAGA | 633

AP3 TCGAGATCAAGCGGATCTAGAACGCGACCAACAGGCAGGTGACCT

Oryza sativa ACTCGAAGCGCCGCACGGGGATCATGAAGAAGG

Giu12Term CCTTCTTCATGATCCCCGTGCGGCGCTTCGAGTAGGTCACCTGCC | 634

GAG-TAG TGTTGGTCGCGTTCTAGATCCGCTTGATCTCGATCTTGCCCCTCCC ® CATGGCCBCCCCCTGCAGCAGCTATCTCTC

AGCGGATCTAGAACGCG

CGCGTTCTAGATCCGCT

Table 15

Oligonucleotides to produce male-sterile plants

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Male-sterile TCTGTACTAATCAAATTTTGCCCTAAACGTTTTTGGCTTTGGAGCA | 637

AG GCAATCACGGCGTAGCAATCGGAGCTAGGAGGAGATTCCTCTCC

® Arabidopsis thaliana | CTTGAGGAAATCTGGGAGAGGAAAGATCGAA

Tyr35Term TTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGAATCTCCT | 638

TAC-TAG CCTAGCTCCGATTGCTACGCCGTGATTGCTGCTCCAAAGCCAAAA

ACGTTTAGGGCAAAATTTGATTAGTACAGA

ACGGCGTAGCAATCGGA

TCCGATTGCTACGCCGT

AG CAATCACGGCGTACTAATCGGAGCTAGGAGGAGATTCCTCTCCCT

Arabidopsis thaliana | TGAGGAAATCTGGGAGAGGAAAGATCGAAA

GIn36Term TTTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGAATCTCC | 642

CAA-TAA TCCTAGCTCCGATTAGTACGCCGTGATTGCTGCTCCAAAGCCAAA

AACGTTTAGGGCAAAATTTGATTAGTACAG

CGGCGTACTAATCGGAG

CTCCGATTAGTACGCCG

111201 01:47 pm

AMENDED SHEET 03137009 — [NY}783550.1

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration

Male-sterile ACTAATCAAATTTTGCCCTAAACGTTTTTGGCTTTGGAGCAGCAAT

AG CACGGCGTACCAATAGGAGCTAGGAGGAGATTCCTCTCCCTTGA

Arabidopsis thaliana | GGAAATCTGGGAGAGGAAAGATCGAAATCAA

Ser37Term TTGATTTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGAAT

TCG-TAG CTCCTCCTAGCTCCTIATTGGTACGCCGTGATTGCTGCTCCAAAGC

CAAAAACGTTTAGGGCAAAATTTGATTAGT

GTACCAATAGGAGCTAG

CTAGCTCCTATTGGTAC

Male-sterie | IAATCAAATTITGCCCTAAACGTTTTTGGCTTTGGAGCAGCAATCA

AG CGGCGTACCAATCGTAGCTAGGAGGAGATTCCTCTCCCTTGAGGA

® Arabidopsis thaliana | AATCTGGGAGAGGAAAGATCGAAATCAAAC

Glu38Term GTTTGATTTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGA

GAG-TAG ATCTCCTCCTAGCTACGATTGGTACGCCGTGATTGCTGCTCCAAA

GCCAAAAACGTTTAGGGCAAAATTTGATTA

ACCAATCGTAGCTAGGA | 651

TCCTAGCTACGATTGGT

Viale-sterile CTCTCCCACTICTTTTCGGIGGTTTATTCATTTGGTGACGATATCA | 653

AG CAGAAGCAATGGATTAAGGTGGGAGTAGTCACGATGCAGAGAGT

Brassica napus AGCAAGAAGATAGGTAGAGGGAAGATAGAGA

Glu3Term TCTCTATCTTCCCTCTACCTATCTTCTTGCTACTCTCTGCATCGTGA | 654

GAA-TAA CTACTGCCACCTTAATCCATTGCTTCTGTGATATCGTCACCAAATG

AATAAACCACCGAAAAGAAGTGGGAGAG

CAATGGATIAAGGTGGG | 655

CCCACCTTAATCCATTG | 656 ® ale-sterile ATTCATTTGGTGACGATATCACAGAAGCAATGGATGAAGGTGGG | 657

AG AGTAGTCACGATGCATAGAGTAGCAAGAAGATAGGTAGAGGGAA

Brassica napus GATAGAGATAAAGAGGATAGAGAACACAACAA

Glu11Term TTGTTGTGTTCTCTATCCTCTTTATCTCTATCTTCCCTCTACCTATC

GAG-TAG TTCTTGCTACTCTATGCATCGTGACTACTCCCACCTTCATCCATTG

CTTCTGTGATATCGTCACCAAATGAATA

ACGATGCATAGAGTAGC

GCTACTCTATGCATCGT | 660

Male-sterile GGTGACGATATCACAGAAGCAATGGATGAAGGTGGGAGTAGTCA | 661

AG CGATGCAGAGAGTAGCTAGAAGATAGGTAGAGGGAAGATAGAGA

Brassica napus TAAAGAGGATAGAGAACACAACAAATCGTCAAG

Lys14Term CTTGACGATTTGTTGTGTTCTCTATCCTCTTTATCTCTATCTTCCCT | 662

AAG-TAG CTACCTATCTTCTAGCTACTCTCTGCATCGTGACTACTCCCACCTT

CATCCATTGCTTCTGTGATATCGTCACC

AGAGTAGCTAGAAGATA

TATCTTCTAGCTACTCT

1412/01 01:47 pm

AMENDED SHEET 03137.003 — [NY}793559.1

28-12-2001 01939797

Phenotype, Gene, . Plant & Targeted Altering Oligos

Alteration

Male-sterile ACGATATCACAGAAGCAATGGATGAAGGTGGGAGTAGTCACGA | 665

AG TGCAGAGAGTAGCAAGTAGATAGGTAGAGGGAAGATAGAGATAAA

Brassica napus GAGGATAGAGAACACAACAAATCGTCAAGTAA

Lys15Term TTACTTGACGATTTGTTGTGTTCTCTATCCTCTTTATCTCTATCTTC

AAG-TAG CCTCTACCTATCTACTTGCTACTCTCTGCATCGTGACTACTCCCAC

CTTCATCCATTGCTTCTGTGATATCGTC

GTAGCAAGTAGATAGGT

ACCTATCTACTTGCTAC

"Male-sterile CAACCAAAAAACTTAAAAA CTCTTTCCITTICCTTACAAGGIGA| 669

AG AGTAATGGACTTCTAAAGTGATCTAACCAGAGAGATCTCACCACAA

@® Lycopersicon AGGAAACTAGGAAGGGGGAAAATTGAGA esculentum TCTCAATTTTCCCCCTTCCTAGTTTCCTTTGTGGTGAGATCTCTCT | 670

Glu4Term GGTTAGATCACTTTAGAAGTCCATTACTTCACCTTGTAAGGAAAGG

CAA-TAA AAAGAGAAGATTTTTAAGTTTTTTGGTTG

TGGACTTCTAAAGTGAT

ATCACTTTAGAAGTCCA

Male-sterile | AAAATCTTCICITTCCTITCCTTACAAGGTGAAGTAATGGACTTCC | 673

AG AAAGTGATCTAACCTGAGAGATCTCACCACAAAGGAAACTAGGAA

Lycopersicon GGGGGAAAATTGAGATCAAAAGGATCGAAA esculentum TTTCGATCCTTTTGATCTCAATTTTCCCCCTTCCTAGTTTCCTTTGT | 674

Arg9Term GGTGAGATCTCTCAGGTTAGATCACTTTGGAAGTCCATTACTTCAC

AGA-TGA CTTGTAAGGAAAGGAAAGAGAAGATTTT

ATCTAACCTGAGAGATC

GATCTCTCAGGTTAGAT

® Male-sterile ATCTTCTCTTTCCTTTCCTTACAAGGTGAAGTAATGGACTTCCAAA | 677

AG GTGATCTAACCAGATAGATCTCACCACAAAGGAAACTAGGAAGGG

Lycopersicon GGAAAATTGAGATCAAAAGGATCGAAAACA esculentum TGTTTTCGATCCTTTTGATCTCAATTTTCCCCCTTCCTAGTTTCCTT | 678

Glu10Term TGTGGTGAGATCTATCTGGTTAGATCACTTTGGAAGTCCATTACTT

GAG-TAG CACCTTGTAAGGAAAGGAAAGAGAAGAT

TAACCAGATAGATCTCA

TGAGATCTATCTGGTTA

: Male-sterile CTITCGTTTCCTTACAAGGTGAAGTAATGGACTTCCAAAGTGATGT

AG AACCAGAGAGATCTGACCACAAAGGAAACTAGGAAGGGGGAAAA

Lycopersicon TTGAGATCAAAAGGATCGAAAACACGACGAA esculentum TTCGTCGTGTTTTCGATCCTTTTGATCTCAATTTTCCCCCTTCCTAG | 682

Ser12Term TTTICCTTTGTGGTCAGATCTCTCTGGTTAGATCACTTTGGAAGTCC

TCA-TGA ATTACTTCACCTTGTAAGGAAAGGAAAG

AGAGATCTGACCACAAA

11/12/01 04:47 pm

AMENDED SHEET 13137.000 ~ [NY]783658.3

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration — Tommomm |W

Male-sterile GTACTCTCTATTTTCATCTTCCAACCCTITCTTICCTTACCAGGTGA

NAG1 AAGTATGGACTTCTAAAGTGATCTAACAAGAGAGATCTCTCCACAA

Nicotiana tabacum AGGAAACTGGGAAGAGGAAAGATTGAGA

GIndTerm TCTCAATCTTTCCTCTTCCCAGTTTCCTTTGTGGAGAGATCTCTCTT| 686

CAA-TAA GTTAGATCACTTTAGAAGTCCATACTTTCACCTGGTAAGGAAAGAA

AGGGTTGGAAGATGAAAATAGAGAGTAC

TGGACTTCTAAAGTGAT

ATCACTTTAGAAGTCCA

® ale-sterile ATCITCCAACCCTTICTTICCTTACCAGGTGAAAGTATGGACTTCC | 689

NAG1 AAAGTGATCTAACATGAGAGATCTCTCCACAAAGGAAACTGGGAA

Nicotiana tabacum GAGGAAAGATTGAGATCAAACGGATCGAAA

Arg9Term TTTCGATCCGTTTGATCTCAATCTTTCCTCTTCCCAGTTTCCTTTGT

AGA-TGA GGAGAGATCTCTCATGTTAGATCACTTTGGAAGTCCATACTTTCAC

CTGGTAAGGAAAGAAAGGGTTGGAAGAT

ATCTAACATGAGAGATC

GATCTCTCATGTTAGAT ~

Male-sterile TTCCAACCCTTTCTTTCCTTACCAGGTGAAAGTATGGACTTCCAAA | 693

NAGH1 GTGATCTAACAAGATAGATCTCTCCACAAAGGAAACTGGGAAGAG

Nicotiana tabacum | GAAAGATTGAGATCAAACGGATCGAAAACA

Glu10Term TGTTTTCGATCCGTTTGATCTCAATCTTTCCTCTTCCCAGTTTCCTT | 694

GAG-TAG TGTGGAGAGATCTATCTTGTTAGATCACTTTGGAAGTCCATACTTT

CACCTGGTAAGGAAAGAAAGGGTTGGAA

TAACAAGATAGATCTCT

® AGAGATCTATCTTGTTA | 696

Male-sterile TTICCTTACCAGGTGAAAGTATGGACTTCCAAAGTGATCTAACAA

NAG1 GAGAGATCTCTCCATAAAGGAAACTGGGAAGAGGAAAGATTGAGA

Nicotiana tabacum TCAAACGGATCGAAAACACAACGAATCGTC

Gin14Term GACGATTCGTTGTGTTTTCGATCCGTTTGATCTCAATCTTITCCTCTT| 698

CAA-TAA CCCAGTTTCCTTTATGGAGAGATCTCTCTTGTTAGATCACTTTGGA

AGTCCATACTTTCACCTGGTAAGGAAAG

TCTCTCCATAAAGGAAA [699

TTTCCTTTATGGAGAGA

Male-sterile GCCTATGAAAACAAACCCAACACGGTCCTGGACGCTGATGCCCAAT 70

AG AGAAGATTGGGAAGGTGAAAGATCGAGATCAAGCGGATCGAAAA

Rosa hybrida CACCACCAATCGTCAAGTCACCTTCTGCAAAA

Gly22Term TTTTGCAGAAGGTGACTTGACGATTGGTGGTGTTTTICGATCCGCT | 702

GGA-TGA TGATCTCGATCTTTCACCTTCCCAATCTTCTTTGGGCATCAGCGTC

CAGGACCGTGTTGGGTTTGTTTTCATAGGC

TGGGAAGGTGAAAGATC

GATCTTTCACCTTCCCA

1112001 04:47 pm

AMENDED SHEET 137.009 NYP93558-1

28-12-2001 01939797

Phenotype, Gene, N } Plant & Targeted Altering Oligos

Alteration fale-sterile TATGAAAACAAACCCAACACGGTCCTGGACGCTGATGCCCAAAGA | 705

AG AGATTGGGAAGGGGATAGATCGAGATCAAGCGGATCGAAAACAC

Rosa hybrida CACCAATCGTCAAGTCACCTTCTGCAAAAGGC

Lys23Term GCCTTTTGCAGAAGGTGACTTGACGATTGGTGGTGTTTTCGATCC | 706

AAG-TAG GCTTGATCTCGATCTATCCCCTTCCCAATCTTCTTTGGGCATCAGC

GTCCAGGACCGTGTTGGGTTTGTTTTCATA

GAAGGGGATAGATCGAG

CTCGATCTATCCCCTTC

Male-sterile AACAAACCCAACACGGTCCTGGACGCTGATGCCCAAAGAAGATTG | 709

AG GGAAGGGGAAAGATCTAGATCAAGCGGATCGAAAACACCACCAA

@ Rosa hybrida TCGTCAAGTCACCTTCTGCAAAAGGCGCAATG

Glu25Term CATTGCGCCTTTTGCAGAAGGTGACTTGACGATTGGTGGTGTTTT | 710

GAG-TAG CGATCCGCTTGATCTAGATCTTTCCCCTTCCCAATCTTCTTTGGGC

ATCAGCGTCCAGGACCGTGTTGGGTTTGTT

GAAAGATCTAGATCAAG

CTTGATCTAGATCTTTC

Male-sterile CCCAACACGGICCTGGACGCTGATGCCCAAAGAAGATTIGGGAAG | 713

AG GGGAAAGATCGAGATCTAGCGGATCGAAAACACCACCAATCGTCA

Rosahybrida ~~ |AGTCACCTTCTGCAAAAGGCGCAATGGTTTGC

Lys27 GCAAACCATTGCGCCTTTTGCAGAAGGTGACTTGACGATTGGTGG | 714

AAG-TAG TGTTTTCGATCCGCTAGATCTCGATCTTTCCCCTTCCCAATCTTCT

TTGGGCATCAGCGTCCAGGACCGTGTTGGG

TCGAGATCTAGCGGATC

GATCCGCTAGATCTCGA

® far GTCTCTAAGCGATTAATCGACCGAGGTATCGCCCGAGAGGAAAAT

Antirrhinum majus CGGGAGAGGAAAGATCGAGATCAAACGGA

Gin7Term TCCGTTTGATCTCGATCTTTCCTCTCCCGATTTTCCTCTCGGGCGA | 718

CAA-TAA TACCTCGGTCGATTAATCGCTTAGAGACGCCATTTCTACTTAGTCA

AAAAGAAAAAAAATAAAAACAGGCAATTG

TAAGCGATTAATCGACC

GGTCGATTAATCGCTTA ale-sterile TTTTIATITITTTICTITTTGACTAAGTAGAAATGGCGTCTCTAAG | 7 far CGATCAATCGACCTAGGTATCGCCCGAGAGGAAAATCGGGAGAG

Antirthinum majus GAAAGATCGAGATCAAACGGATCGAAAACA

Glu10Term TGTTTTCGATCCGTTTGATCTCGATCTTTCCTCTCCCGATTTTCCTC | 722 © 25 GAG-TAG TCGGGCGATACCTAGGTCGATTGATCGCTTAGAGACGCCATTTCT

ACTTAGTCAAAAAGAAAAAAAATAAAAAC

AATCGACCTAGGTATCG

CGATACCTAGGTCGATT

111201 01:47 pm

AMENDED SHEET 03137.009 — [NY]793559.1 e———— 28-12-2001 01939797

Phenotype, Gene, . Plant & Targeted Altering Oligos

Alteration

Male-sterile TTTCTTTTTGACTAAGTAGAAATGGCGTCTCTAAGCGATCAATCGA | 725 far CCGAGGTATCGCCCTAGAGGAAAATCGGGAGAGGAAAGATCGAG

Antirrhinum majus ATCAAACGGATCGAAAACAAAACAAATCAAC

Glu14Term GTTGATTTGTTTTGTTTTCGATCCGTTTGATCTCGATCTTTCCTCTC | 726

GAG-TAG CCGATTTTCCTCTAGGGCGATACCTCGGTCGATTGATCGCTTAGA

GACGCCATTTCTACTTAGTCAAAAAGAAA

TATCGCCCTAGAGGAAA

TTTCCTCTAGGGCGATA far GTATCGCCCGAGAGGTAAATCGGGAGAGGAAAGATCGAGATCAA ® Antirrhinum majus ACGGATCGAAAACAAAACAAATCAACAGGTTA

Lys16Term TAACCTGTTGATTTGTTTTGTTTTCGATCCGTTTGATCTCGATCTTT | 730

AAA-TAA CCTCTCCCGATTTACCTCTCGGGCGATACCTCGGTCGATTGATCG

CTTAGAGACGCCATTTCTACTTAGTCAAA

CCGAGAGGTAAATCGGG

CCCGATTTACCTCTCGG

Male-sterile | TGTCCAAGCATTAICAGTCACCACTCACAAGAATGATTAAGGAAGA| 733

AG AGGAAAGGGTAAGTAGCAAATAAAGGGGATGTTCCAGAATCAAGA

Cucumis sativus AGAGAAGATGTCAGACTCGCCTCAGAGGAA

Leu21Term TTCCTCTGAGGCGAGTCTGACATCTTCTCTTCTTGATTCTGGAACA | 734

TTG-TAG TCCCCTTTATTTGCTACTTACCCTTTCCTTCTTCCTTAATCATTCTT

GTGAGTGGTGACTGATAATGCTTGGACA

GGGTAAGTAGCAAATAA

TTATTTGCTACTTACCC

®

AG GGAAAGGGTAAGTTGTAAATAAAGGGGATGTTCCAGAATCAAGAA

Cucumis sativus GAGAAGATGTCAGACTCGCCTCAGAGGAAGA

Gin22Term TCTTCCTCTGAGGCGAGTCTGACATCTTCTCTTCTTGATTCTGGAA | 738

CAA-TAA CATCCCCTTTATTTACAACTTACCCTTTCCTTCTTCCTTAATCATTC

TTGTGAGTGGTGACTGATAATGCTTGGA

GTAAGTTGTAAATAAAG

CTTTATTTACAACTTAC

Male-sterile CATTATCAGTCACCACTCACAAGAATGATTAAGGAAGAAGGAAAG | 741

AG GGTAAGTTGCAAATATAGGGGATGTTCCAGAATCAAGAAGAGAAG

Cucumis sativus ATGTCAGACTCGCCTCAGAGGAAGATGGGAA

Lys24 Term TTCCCATCTTCCTCTGAGGCGAGTCTGACATCTTCTCTTCTTGATT | 742 - 25 AAG-TAG CTGGAACATCCCCTATATTTGCAACTTACCCTTTCCTTCTTCCTTAA

TCATTCTTGTGAGTGGTGACTGATAATG

TGCAAATATAGGGGATG

CATCCCCTATATTTGCA

1112/01 01:47 pm

AM EN DED S H EET 3437.009 — [NY)783559.1

28-12-2001 01939797

Phenotype, Gene, } Plant & Targeted Altering Oligos

Alteration ale-sterile CCACTCACAAGAATGATTAAGGAAGAAGGAAAGGGTAAGTTGCAA | 745

AG ATAAAGGGGATGTTCTAGAATCAAGAAGAGAAGATGTCAGACTCG

Cucumis sativus CCTCAGAGGAAGATGGGAAGAGGAAAGATTG

Gln28Term CAATCTTTCCTCTTCCCATCTTCCTCTGAGGCGAGTCTGACATCTT | 746

CAG-TAG CTCTTCTTGATTCTAGAACATCCCCTTTATTTGCAACTTACCCTTTC

CTTCTTCCTTAATCATTCTTGTGAGTGG

GGATGTTCIAGAATCAA

TTGATTCTAGAACATCC

Male-sterile CCACCACCACCACCACCACCACCACCACACCATGCTCAACATGAT | 749

AG GACTGATCTGAGCTGAGGGCCGTCGTCCAAGGTCAAGGAGCAGG

® Zea mays TGGCGGCGGCGCCGACGGGCTCCGGCGACAGS

Cys10Term CCTGTCGCCGGAGCCCGTCGGCGCCGCCGCCACCTGCTCCTTGA | 750

TGC-TGA CCTTGGACGACGGCCCTCAGCTCAGATCAGTCATCATGTTGAGCA

TGGTGTGGTGGTGGTGGTGGTGGTGGTGGTIGG

CTGAGCTGAGGGCCGTC

GACGGCCCICAGCTCAG

Male-sterile ACCACCACCACCACCACCACACCATGCTCAACATGATGACTGA 753

AG TGAGCTGCGGGCCGTAGTCCAAGGTCAAGGAGCAGGTGGCGGC

Zea mays GGCGCCGACGGGCTCCGGCGACAGGCAGGGGCA

Ser13Tem TGCCCCTGCCTGTCGCCGGAGCCCGTCGGCGCCGCCGCCACCT | 754

TCG-TAG GCTCCTTGACCTTGGACTACGGCCCGCAGCTCAGATCAGTCATCA

TGTTGAGCATGGTGTGGTGGTGGTGGTGGTGGT

CGGGCCGTAGTCCAAGG

CCTTGGACTACGGCCCG

® Male-sterile CACCACCACCACCACACCATGCTCAACATGATGACTGATCTGAGC | 757

AG TGCGGGCCGTCGTCCTAGGTCAAGGAGCAGGTGGCGGCGGCGC

Zea mays CGACGGGCTCCGGCGACAGGCAGGGGCAGGGGA

Lys15Term TCCCCTGCCCCTGCCTGTCGCCGGAGCCCGTCGGCGCCGCCGE | 758

AAG-TAG CACCTGCTCCTTGACCTAGGACGACGGCCCGCAGCTCAGATCAG

TCATCATGTTGAGCATGGTGTGGTGGTGGTGGTG

CGTCGTCCTAGGTCAAG

CTTGACCTAGGACGACG

Male-sterile CACCACCACACCATGCTCAACATGATGACTGATCTGAGCTGCGGG | 76

AG CCGTCGTCCAAGGTCTAGGAGCAGGTGGCGGCGGCGCCGACGG

Zea mays GCTCCGGCGACAGGCAGGGGCAGGGGAGAGGCA

Lys17Term TGCCTCTCCCCTGCCCCTGCCTGTCGCCGGAGCCCGTCGGCGCE | 762 + 25 AAG-TAG GCCGCCACCTGCTCCTAGACCTTGGACGACGGCCCGCAGCTCAG

ATCAGTCATCATGTTGAGCATGGTGTGGTGGTG

CCAAGGTCTAGGAGCAG

CTGCTCCTAGACCTTGG

1112101 01:47 pm

AMENDED SHEET 137.005 — [NYI793553.1

28-12-2001 01939797

SE N_—

Phenotype, Gene, . Plant & Targeted Altering Oligos

Alteration

Malesterle | TCCTACCTITICTCCTTCAGACCTCAAAATCTGTGTGATAGGAACA | 765

AG AGAGCATGCACATCTGAGAAGAGGAGGCTACACCATCCACAGTAA

Zea mays CAGGCATCATGTCGACCCTGACTTCGGCGG

ArgdTerm CCGCCGAAGTCAGGGTCGACATGATGCCTGTTACTGTGGATGGT | 766

CGA-TGA GTAGCCTCCTCTTCTCAGATGTGCATGCTCTTGTTCCTATCACACA

GATTTTGAGGTCTGAAGGAGAAAAGGTAGGA

TGCACATCTGAGAAGAG

CTCTTCTCAGATGTGCA

Male-sterile TACCTTT1GTCCTTCAGACCTCAAAATCTGTGTGATAGGAACAAGA | 769

AG GCATGCACATCCGATAAGAGGAGGCTACACCATCCACAGTAACAG

® Zea mays GCATCATGTCGACCCTGACTTCGGCGGGGC

GlusTerm GCCCCGCCGAAGTCAGGGTCBACATGATGCCTGTTACTGTGGAT | 770

GAA-TAA GGTGTAGCCTCCTCTTATCGGATGTGCATGCTCTTGTTCCTATCAC

ACAGATTTTGAGGTCTGAAGGAGAAAAGGTA

ACATCCGATAAGAGGAG

CTCCTCTTATCGGATGT | 772

Male-sterile CTTTTCTCCTTCAGACCTCAAAATCTGTGTGATAGGAACAAGAGCA | 773

AG TGCACATCCGAGAATAGGAGGCTACACCATCCACAGTAACAGGCA

Zea mays TCATGTCGACCCTGACTTCGGCGGGGCAGC

GlugTerm GCTGCCCCGCCGAAGTCAGGGTCGACATGATGCCTGTTACTGTG | 774 16 GAG-TAG GATGGTGTAGCCTCCTATTCTCGGATGTGCATGCTCTTGTTCCTAT

CACACAGATTTTGAGGTCTGAAGGAGAAAAG

TCCGAGAATAGGAGGCT

AGCCTCCTATTCTCGGA ee ® Male-sterile CTCCTTCAGACCTCAAAATCTGTGTGATAGGAACAAGAGCATG | 777

AG CACATCCGAGAAGAGTAGGCTACACCATCCACAGTAACAGGCATC

Zea mays ATGTCGACCCTGACTTCGGCGGGGCAGCAGA

Glu7Term TCTGCTGCCCCGCCGAAGTCAGGGTCGACATGATGCCTGTTACT | 778

GAG-TAG GTGGATGGTGTAGCCTACTCTTCTCGGATGTGCATGCTCTTGTTC

CTATCACACAGATTTTGAGGTCTGAAGGAGAA

GAGAAGAGTAGGCTACA

TGTAGCCTACTCTTCTC ee

Male-sterile TGGGTCAGGATCGTCGGCGGCGGTGGCGGCGGGGAGCAGC | 78

AG GAGAAGATGGGGAGGGGGTAGATCGAGATAAAGCGGATCGAGAA

Oryza sativa CACGACGAACCGGCAGGTGACCTTCTGCAAGCGCC

Lys5Term GGCGCTTGCAGAAGGTCACCTGCCGGTTCGTCGTGTTCTCGATC | 782 © 25 AAG-TAG CGCTTTATCTCGATCTACCCCCTCCCCATCTTCTCGCTGCTCCCC

GCCGCCACCGCCGCCGACGATCCTGACCCAGC

GGAGGGGGTAGATCGAG

CTCGATCTACCCECTCC | 784 |] 11/12/01 01:47 pm

AMENDED SHEET 3137.009 ~ [NY]793559.1

Eee — 28-12-2001 01939797

Phenotype, Gene, } Plant & Targeted Altering Oligos

Alteration

Male-sterile TCAGGATCGTCGGCGGCGGTGGCGGCGGGGAGCAGCGAGAAGA | 785

AG TGGGGAGGGGGAAGATCTAGATAAAGCGGATCGAGAACACGACG

Oryza sativa AACCGGCAGGTGACCTTCTGCAAGCGCCGCAATG

Glu7Term CATTGCGGCGCTTGCAGAAGGTCACCTGCCGGTTCGTCGTGTTCT | 786

GAG-TAG CGATCCGCTTTATCTAGATCTTCCCCCTCCCCATCTTCTCGCTGCT

CCCCGCCGCCACCGCCGCCGACGATCCTGA

GGAAGATCTAGATAAAG

____ |CTITATCTAGATCTTCC | 788

Male-sterile TCGTCGGCGGCGGTGGCGGCGGGGAGCAGCGAGAAGATGGGG 789

AG AGGGGGAAGATCGAGATATAGCGGATCGAGAACACGACGAACCG

@® Oryza sativa GCAGGTGACCTTCTGCAAGCGCCGCAATGGCCTCC

Lys9Term GGAGGCCATTGCGGCGCTTGCAGAAGGTCACCTGCCGGTTCGTC | 790

AAG-TAG GTGTTCTCGATCCGCTATATCTCGATCTTCCCCCTCCCCATCTTCT

CGCTGCTCCCCGCCGCCACCGCCGCCGACGA

TCGAGATATAGCGGATC

GATCCGCTATATCTCGA ale-sterile GCGGTGGCGGCGGGGAGCAGCGAGAAGATGGGGAGGGGGAAG | 79

AG ATCGAGATAAAGCGGATCTAGAACACGACGAACCGGCAGGTGAC

Oryza sativa CTTCTGCAAGCGCCGCAATGGCCTCCTGAAGAAGG

Glu12Term CCTTCTTCAGGAGGCCATTGCGGCGCTTGCAGAAGGTCACCTGC 794

GAG-TAG CGGTTCGTCGTGTTCTAGATCCGCTTTATCTCGATCTTCCCCCTCC

CCATCTTCTCGCTGCTCCCCGCCGCCACCGC

AGCGGATCTAGAACACG

CGTGTTCTAGATCCGCT

Table 16

Oligonucleotides to produce male-sterile plants

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration .

PI TAGACAAGTTACATAGTCAAAGAGAAGAAATGGTATCATCAAAAAA

Cucumis sativus GCCAAAGAAATTACTGTTCTTTGCGATGCT ’ Tyr21Term AGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATGATACCAT | 798

TAT-TAG TTCTTCTCTTTGACTATGTAACTTGTCTATTGCTTGAGTTCTCTATTC ____ ITTTTTATITCTATTITCCCTCTTCCC 111201 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene, B

Plant & Targeted Altering Oligos ’ Alteration

GTTACATAGTCAAAGAG

CTCTTTGACTATGTAAC

AAAATAGAAATAAAAAGAATAGAGAACTCAAGCAATA | 801

Pl GACAAGTTACATATTGAAAGAGAAGAAATGGTATCATCAAAAAAGC

Cucumis sativus CAAAGAAATTACTGTTCTTTGCGATGCTCA

Ser22Term TGAGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATGATAC | 802

TCA-TGA CATTTCTTCTCTTTCAATATGTAACTTGTCTATTGCTTGAGTTCTCTA

TTCTTTTTATTTCTATTTTCCCTCTTC

TACATATTGAAAGAGAA

¢ TTCTCTTTCAATATGTA

Male-sterile AGAGGGAAAATAGAAATAAAAAGAATAGAGAACTCAAGCAATAGAC

Pi AAGTTACATATTCATAGAGAAGAAATGGTATCATCAAAAAAGCCAA

Cucumis sativus AGAAATTACTGTTCTTTGCGATGCTCAAG

Lys23Term CTTGAGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATGATA| 806

AAG-TAG CCATTTCTTCTCTATGAATATGTAACTTGTCTATTGCTTGAGTTCTC

TATTCTTTTTATTTCTATTTTCCCTCT

CATATTCATAGAGAAGA

TCTTCTCTATGAATATG

Male-sterile GGGAAAATAGAAATAAAAAGAATAGAGAACTCAAGCAATAGACAAG| 809

PI TTACATATTCAAAGTGAAGAAATGGTATCATCAAAAAAGCCAAAGA

Cucumis sativus AATTACTGTTCTTTGCGATGCTCAAGTTT

Arg24Term AAACTTGAGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATG| 810

AGA-TGA ATACCATTTCTTCACTTTGAATATGTAACTTGTCTATTGCTTGAGTT

CTCTATTCTTTTTATTTCTATTTTCCC

® ATTCAAAGTGAAGAAAT

ATTTCTTCACTTTGAAT

PI CAGGCAGGTGACCTAGTCCAAGAGGAGGAATGGGATTATCAAGAA

Malus domestica GGCAAAGGAGATCACTGTTCTATGTGATGCT

Tyr21Term AGCATCACATAGAACAGTGATCTCCTTTGCCTTCTTGATAATCCCA | 814

TAC-TAG TTCCTCCTCTTGGACTAGGTCACCTGCCTGTTACTTGAGTTCTCAA

TCCTCTTGATCTCAACCTTCCCACGTCCC

GTGACCTAGTCCAAGAG

CTCTTGGACTAGGTCAC

1112/01 01:47 pm 03137.009 — [NY)793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

Male-sterile CGTGGGAAGGTTGAGATCAAGAGGATTGAGAACTCAAGTAACAGG | 817

Pl CAGGTGACCTACTCCTAGAGGAGGAATGGGATTATCAAGAAGGCA

Malus domestica AAGGAGATCACTGTTCTATGTGATGCTAAAG

Lys23Term CTTTAGCATCACATAGAACAGTGATCTCCTTTGCCTTCTTGATAATC| 818

AAG-TAG CCATTCCTCCTCTAGGAGTAGGTCACCTGCCTGTTACTTGAGTTCT

CAATCCTCTTGATCTCAACCTTCCCACG

CCTACTCCTAGAGGAGG | 819

CCTCCTCTAGGAGTAGG

Male-sterile AGGATTGAGAACTCAAGTAACAGGCAGGTGACCTACTCCAAGAGG | 821

Pl AGGAATGGGATTATCTAGAAGGCAAAGGAGATCACTGTTCTATGT ® Malus domestica GATGCTAAAGTATCTCTTATCATTTATTCTA

Lys30Term TAGAATAAATGATAAGAGATACTTTAGCATCACATAGAACAGTGAT | 822

AAG-TAG CTCCTTTGCCTTCTAGATAATCCCATTCCTCCTCTTGGAGTAGGTC

ACCTGCCTGTTACTTGAGTTCTCAATCCT

GGATTATCTAGAAGGCA

TGCCTTCTAGATAATCC

Male-sterile ATTGAGAACTCAAGTAACAGGCAGGTGACCTACTCCAAGAGGAGG | 825

PI AATGGGATTATCAAGTAGGCAAAGGAGATCACTGTTCTATGTGATG

Malus domestica CTAAAGTATCTCTTATCATTTATTCTAGCT

Lys31Term AGCTAGAATAAATGATAAGAGATACTTTAGCATCACATAGAACAGT | 826

AAG-TAG GATCTCCTTTGCCTACTTGATAATCCCATTCCTCCTCTTGGAGTAG

GTCACCTGCCTGTTACTTGAGTTCTCAAT

TTATCAAGTAGGCAAAG

CTTTGCCTACTTGATAA

@® globosa AAACAAAAAAATGTGAAGAGGAAAAATTGAGATCAAAAGAATTGAG CC

Antirthinum majus AACTCAAGCAACAGGCAGGTTACTTACT

Gly2Term AGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTCTTTTGATCTCA| 830

GGA-TGA ATTTTTCCTCTTCACATTTTTTTGTTTTTGTTITTICTCTCTTGTTTTTG

TTTGCAGATAACTATTGTAAAAATG

AAAAAATGTGAAGAGGA

TCCTCTTCACATTTIIT

AAAAACAAGAGAGAAAAACAAAAA] 833 globosa CAAAAAAATGGGATGAGGAAAAATTGAGATCAAAAGAATTGAGAAC

Antirrhinum majus TCAAGCAACAGGCAGGTTACTTACTCAA : Arg3Term TTGAGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTCTTITGATC | 834

AGA-TGA TCAATTTTTCCTCATCCCATTTTTITTGTTTTIGTTTTTCTCTCTTGTTT

TTGTTTGCAGATAACTATTGTAAAA

1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ; Alteration

AAATGGGATGAGGAAAA

: TTTTCCTCATCCCATTT

Male-sterile TACAATAGTTATCTGCAAACAAAAACAAGAGAGAAAAACAAAAACA | 837 globosa AAAAAATGGGAAGATGAAAAATTGAGATCAAAAGAATTGAGAACTC

Antirrhinum majus AAGCAACAGGCAGGTTACTTACTCAAAGA

Gly4Term TCTTTGAGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTCTTTTG | 838

GGA-TGA ATCTCAATTTTTCATCTTCCCATTTTTTTGTTTTTGTTTTTCTCTCTTG

TTTTTGTTTGCAGATAACTATTGTA

TGGGAAGATGAAAAATT

® AATTTTTCATCTTCCCA

Male-sterile AATAGTTATCTGCAAACAAAAACAAGAGAGAAAAACAAAAACAAAA | 841 globosa AAATGGGAAGAGGATAAATTGAGATCAAAAGAATTGAGAACTCAAG

Antirrhinum majus CAACAGGCAGGTTACTTACTCAAAGAGAA

Lys5Term TTCTCTTTGAGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTCTT | 842

AAA-TAA TTGATCTCAATTTATCCTCTTCCCATTTTTTTGTTTTTGTTTITCTCT

CTTGTTTTTGTTTGCAGATAACTATT

GAAGAGGATAAATTGAG

CTCAATTTATCCTCTTC

Viale-steriie A : A AGAA of

PI AGTATGGGGCGCGGCTAGATCAAGATCAAGAGGATCGAGAACTCT

Zea mays ACCAACCGGCAGGTGACCTTCTCCAAGCGCC

Lys5Term GGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTCTCGATCC| 846

AAG-TAG TCTTGATCTTGATCTAGCCGCGCCCCATACTGCGTTCTCCACTCCC

AAACAGATCCAAGGGCAGCAAGAGCTCAGC

® GGCGCGGCTAGATCAAG

CTTGATCTAGCCGCGCC

Male-sterile CTCTTGCTGCCCTTGGATCTGTTTGGGAGTGGAGAACGCAGTATG | 849

Pl GGGCGCGGCAAGATCTAGATCAAGAGGATCGAGAACTCTACCAAC

Zea mays CGGCAGGTGACCTTCTCCAAGCGCCGGGCCG

Lys7Term CGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTC

AAG-TAG TCGATCCTCTTGATCTAGATCTTGCCGCGCCCCATACTGCGTTCTC

CACTCCCAAACAGATCCAAGGGCAGCAAGAG

GCAAGATCTAGATCAAG

CTTGATCTAGATCTTGC

1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797 -03.

Phenotype, Gene,

Plant & Targeted Altering Oligos " Alteration

Male-sterile CTCTTGCTGCCCTTGGATCTGTTTGGGAGTGGAGAACGCAGTATG | 853

P) GGGCGCGGCAAGATCTAGATCAAGAGGATCGAGAACTCTACCAAC

Zea mays CGGCAGGTGACCTTCTCCAAGCGCCGGGCCG

Lys9Term CGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTIC | 854

AAG-TAG TCGATCCTCTTGATCTAGATCTTGCCGCGCCCCATACTGCGTTCTC

CACTCCCAAACAGATCCAAGGGCAGCAAGAG

GCAAGATCTAGATCAAG

CTTGATCTAGATCTTGC

Pi CAAGATCAAGAGGATCTAGAACTCTACCAACCGGCAGGTGACCTT @® Zea mays CTCCAAGCGCCGGGCCGGACTGGTCAAGAAGG

Glu12Term CCTTCTTGACCAGTCCGGCCCGGCGCTTGGAGAAGGTCACCTGC | 858

GAG-TAG CGGTTGGTAGAGTTCTAGATCCTCTTGATCTTGATCTTGCCGCGCC

CCATACTGCGTTCTCCACTCCCAAACAGATC

AGAGGATCTAGAACTCT | 859

AGAGTTCTAGATCCTCT

Viale-sterile GA AA ; A AGAL 55

PI AGTATGGGGCGCGGCTAGATCGAGATCAAGAGGATCGAGAACTCT

Zea mays ACCAACCGGCAGGTGACCTTCTCCAAGCGCC

Lys5Term GGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTCTCGATCC| 862

AAG-TAG TCTTGATCTCGATCTAGCCGCGCCCCATACTCCGTTCTCCACTCCC

TAACAGATTCAAGGGCAGCAAGAGCTCAGC

GGCGCGGCTAGATCGAG

CTCGATCTAGCCGCGCC

® Male-sterile CTCTTGCTGCCCTTGAATCTGTTAGGGAGTGGAGAACGGAGTATG

Pl GGGCGCGGCAAGATCTAGATCAAGAGGATCGAGAACTCTACCAAC

Zea mays CGGCAGGTGACCTTCTCCAAGCGCCGGGCCG

Glu7Term CGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTC | 866

GAG-TAG TCGATCCTCTTGATCTAGATCTTGCCGCGCCCCATACTCCGTTCTC

CACTCCCTAACAGATTCAAGGGCAGCAAGAG

GCAAGATCTAGATCAAG

CTTGATCTAGATCTTGC

Male-sterile CTGCCCTTGAATCTGTTAGGGAGTGGAGAACGGAGTATGGGGCG | 869 g Pl CGGCAAGATCGAGATCTAGAGGATCGAGAACTCTACCAACCGGCA

Zea mays GGTGACCTTCTCCAAGCGCCGGGCCGGACTGG

Lys9Term CCAGTCCGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTA | 670

AAG-TAG GAGTTCTCGATCCTCTAGATCTCGATCTTGCCGCGCCCCATACTC

CGTTCTCCACTCCCTAACAGATTCAAGGGCAG

11/12/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene, }

Plant & Targeted Altering Oligos . Alteration

TCGAGATCTAGAGGATC

: GATCCTCTAGATCTCGA

Male-sterile AATCTGTTAGGGAGTGGAGAACGGAGTATGGGGCGCGGCAAGAT | 873

Pl CGAGATCAAGAGGATCTAGAACTCTACCAACCGGCAGGTGACCTT

Zea mays CTCCAAGCGCCGGGCCGGACTGGTCAAGAAGG

Glui2Term CCTTCTTGACCAGTCCGGCCCGGCGCTTGGAGAAGGTCACCTGC | 874

GAG-TAG CGGTTGGTAGAGTTCTAGATCCTCTTGATCTCGATCTTGCCGCGC

CCCATACTCCGTTCTCCACTCCCTAACAGATT

AGAGGATCTAGAACTCT

® AGAGTTCTAGATCCTCT 'Male-s er 2) AA A A AA /\ /\ /\ A A A 3

Pi CGGGATGGCGCGCGGGTAGATCGAGATCAAGAGGATCGAGAACT

Oryza sativa CCACCAACCGCCAGGTGACCTTCTCCAAGCGCA

Lys5Term TGCGCTTGGAGAAGGTCACCTGGCGGTTGGTGGAGTTCTCGATCC| 878

AAG-TAG TCTTGATCTCGATCTACCCGCGCCCCATCCCGCCTCCTCCTCCTC

CTCCTCCTTCCTCCAGCTAGCTTAGCAGCAA

GGCGCGGGTAGATCGAG

CTCGATCTACCCGCGCC

Male-sterile CTAAGCTAGCTGGAGGAAGGAGGAGGAGGAGGAGGAGGCGGGA | 881

PI TGGGGCGCCGGAAGATCTAGATCAAGAGGATCGAGAACTCCACC

Oryza sativa AACCGCCAGGTGACCTTCTCCAAGCGCAGGAGCG

Glu7Term CGCTCCTGCGCTTGGAGAAGGTCACCTGGCGGTTGGTGGAGTTCT| 882

GAG-TAG CGATCCTCTTGATCTAGATCTTCCCGCGCCCCATCCCGCCTCCTC

CTCCTCCTCCTCCTTCCTCCAGCTAGCTTAG

® GGAAGATCTAGATCAAG

CTTGATCTAGATCTTCC

Male-sterile TAGCTGGAGGAAGGAGGAGGAGGAGGAGGAGGCGGGATGGGGC | 885

PI GCGGGAAGATCGAGATCTAGAGGATCGAGAACTCCACCAACCGC

Oryza sativa CAGGTGACCTTCTCCAAGCGCAGGAGCGGGATCC

Lys9Term GGATCCCGCTCCTGCGCTTGGAGAAGGTCACCTGGCGGTTGGTG | 886

AAG-TAG GAGTTCTCGATCCTCTAGATCTCGATCTTCCCGCGCCCCATCCCG

CCTCCTCCTCCTCCTCCTCCTTCCTCCAGCTA

TCGAGATCTAGAGGATC

GATCCTCTAGATCTCGA

11/12/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene, :

Plant & Targeted Altering Oligos .

Male-sterile GAAGGAGGAGGAGGAGGAGGAGGCGGGATGGGGCGCGGGAAG | 889 . PI ATCGAGATCAAGAGGATCTAGAACTCCACCAACCGCCAGGTGACC

Oryza sativa TTCTCCAAGCGCAGGAGCGGGATCCTCAAGAAGG

Glut2Term CCTTCTTGAGGATCCCGCTCCTGCGCTTGGAGAAGGTCACCTGGC | 890

GAG-TAG GGTTGGTGGAGTTCTAGATCCTCTTGATCTCGATCTTCCCGCGCC

CCATCCCGCCTCCTCCTCCTCCTCCTCCTTC

AGAGGATCTAGAACTCC

GGAGTTCTAGATCCTCT

14/42/01 01:47 pm 03137.000 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Example 7

Engineering plants for abiotic stress tolerance

Environmental stresses, such as drought, increased soil salinity, soil contamination with heavy metals, and extreme temperature, are major factors limiting plant growth and productivity. The worldwide loss in yield of three major cereal crops, rice, maize, and wheat due to water stress (drought) has been estimated to be over ten billion dollars annually and many currently marginal soils could be brought into cultivation if suitable plant varieties were available.

Physiological and biochemical responses to high levels of ionic or nonionic solutes and ® decreased water potential have been studied in a variety of plants. It is known, for example, that increasing levels of alcohol dehydrogenase can confer enhances flooding resistance in plants. There are also several possible mechanisms to enhance plant salt tolerance. For example, one mechanism underlying the adaptation or tolerance of plants to osmotic stresses is the accumulation of compatible, low molecular weight osmolytes such as sugar alcohols, special amino acids, and glycinebetaine. Such accumulation can be engineered, for example, by removing feedback inhibition on 1-pyrroline-t-carboxylate synthetase, which 16 results in accumulation of proline. Additionally, recent experiments suggest that altering the expression or activity of specific sodium or potassium transporters can confer enhanced salt tolerance.

Plant tolerance of contamination by heavy metals such as lead and aluminum in soils has also been investigated and one mechanism underlying tolerance is the production of dicarboxylic acids such as oxalate and citrate. In addition, individual genes involved in heavy metal sensitivity have been identified. ® The attached tables disclose exemplary oligonucleotide base sequences which can be used to generate site-specific mutations that confer stress tolerance in plants.

OL LC MRL Sik

AMENDED SHEET

28-12-2001 01939797

Table 17

J Genome-Altering Oligos Conferring Stress Tolerance

In Phenotype, Gene, . Plant & Targeted Altering Oligos

Salt Tolerance CGTCTTTTTGTGTGGTAGTTGGATGTGACGGTTGCTCAAATGCTT | 893

P5CS GTGACCGATAGCAGTGCTAGAGATAAGGATTTCAGGAAGCAACTT

Arabidopsis thaliana | AGTGAAACTGTCAAAGCGATGCTGAGGATGA

Phe128Ala TCATCCTCAGCATCGCTTTGACAGTTTCACTAAGTTGCTTCCTGAA | 894

TTT-GCT ATCCTTATCTCTAGCACTGCTATCGGTCACAAGCATTTGAGCAACC

GTCACATCCAACTACCACACAAAAAGACG

ATAGCAGTGCTAGAGAT

® ATCTCTAGCACTGCTAT

Salt Tolerance GAGACTATGTTTGACCAGCTGGATGTGA GCTCAGCTGCIG| 897

P5CS 1 GTGAATGACAGTAGTGCCAGAGACAAGGAGTTCAGGAAGCAACTT

Brassica napus AATGAGACAGTGAAGTCCATGCTTGATTTGA

Phe128Ala TCAAATCAAGCATGGACTTCACTGTCTCATTAAGTTGCTTCCTGAA | 898

TTC-GCC CTCCTTGTCTCTGGCACTACTGTCATTCACCAGCAGCTGAGCAGC

CGTCACATCCAGCTGGTCAAACATAGTCTC

ACAGTAGTGCCAGAGAC

GTCTCTGGCACTACTGT 900

Salt Tolerance GAGACTATGTTTGACCAGATGGATGTGACGGTGGCTCAAATGCTG

P5CS 2 GTGACTGATAGCAGTGTCAGAGATAAGGATTTCAGGAAGCAACTT

Brassica napus AGTGAGACAGTCAAAGCTATGCTGAAAATGA

Phe129Ala TCATTTTCAGCATAGCTTTGACTGTCTCACTAAGTTGCTTCCTGAA | 902

TTC-GCC ATCCTTATCTCTGACACTGCTATCAGTCACCAGCATTTGAGCCACC

PS GTCACATCCATCTGGTCAAACATAGTCTC

ATAGCAGTGTCAGAGAT

ATCTCTGACACTGCTAT | 904

Salt Tolerance GATATGTTGTTTAACCAACTGGATGTCTCGTCATCTCAACTTCTTG | 905

P5CS TCACCGACAGTGATGCTGAGAACCCAAAGTTCCGGGAGCAACTCA

Oryza sativa CTGAAACTGTTGAGTCATTATTAGATCTTA

Phe128Ala TAAGATCTAATAATGACTCAACAGTTTCAGTGAGTTGCTCCCGGAA

TTT-GCT CTTTGGGTTCTCAGCATCACTGTCGGTGACAAGAAGTTGAGATGA ; CGAGACATCCAGTTGGTTAAACAACATATC a ACAGTGATGCTGAGAAC

GTTCTCAGCATCACTGT

11/12/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos © Alteration

Salt Tolerance GATATTTTGTTTAGTCAGCTGGATGTGACATCTGCTCAGCTTCTTG

P5CS TTACTGACAATGATGCTAGAGACCAAGATTTTAGAAAGCAACTTTC

Medicago sativa TGAAACTGTGAGATCACTTCTAGCACTAA

Phe128Ala TTAGTGCTAGAAGTGATCTCACAGTTTCAGAAAGTTGCTTTCTAAA | 910

TTT-GCT ATCTTGGTCTCTAGCATCATTGTCAGTAACAAGAAGCTGAGCAGAT

GTCACATCCAGCTGACTAAACAAAATATC

ACAATGATGCTAGAGAC

GTCTCTAGCATCATTGT

Salt Tolerance GATACATTGTTTAGTCAGCTGGATGTGACATCAGCTCAGCTACTC | 913

P5CS GTTACTGATAATGATGCTAGGGATCCAGAATTCAGGAAGCAACTT ® Actinidia deliciosa ACTGAAACTGTAGAATCACTATTGAATTTGA

Phe128Ala TCAAATTCAATAGTGATTCTACAGTTTCAGTAAGTTGCTTCCTGAAT | 914

TTT-GCT TCTGGATCCCTAGCATCATTATCAGTAACGAGTAGCTGAGCTGAT

GTCACATCCAGCTGACTAAACAATGTATC

ATAATGATGCTAGGGAT

ATCCCTAGCATCATTAT ]

Salt Tolerance GACACACTCTTCAGTCAACTGGATGTGACATCAGCACAGCTTCTT | 917

P5CS GTAACAGATAATGACGCCAGAAGTCCAGAATTTAGAAAACAACTTA

Cichorium intybus CTGAAACAGTCGATTCTTTATTATCTTATA

Phe122Ala TATAAGATAATAAAGAATCGACTGTTTCAGTAAGTTGTTTTCTAAAT | 918

TTC-GCC TCTGGACTTCTGGCGTCATTATCTGTTACAAGAAGCTGTGCTGAT

GTCACATCCAGTTGACTGAAGAGTGTGTC

ATAATGACGCCAGAAGT 919

ACTTCTGGCGTCATTAT

® Salt Tolerance GATTCTITGITCAGTCAGTTGGATGTGACATCAGCTCAGCTTCTGG [ 921

P5CS TGACTGATAATGACGCTAGAGATCCAGATTTTAGGAGACAACTCA

Lycopersicon ATGACACAGTAAATTCGTTGCTTTCTCTAA esculentum TTAGAGAAAGCAACGAATTTACTGTGTCATTGAGTTGTCTCCTAAA | 922

Phe128Ala ATCTGGATCTCTAGCGTCATTATCAGTCACCAGAAGCTGAGCTGA

TTT-GCT TGTCACATCCAACTGACTGAACAAAGAATC

ATAATGACGCTAGAGAT

ATCTCTAGCGTCATTAT alt Tolerance ATACCATGTTCAGCCAGCTTGATGTGACTTCT AACTTCTTG | 925 - P5CS TGAATGATGGATTTGCTAGGGATGCTGGCTTCAGAAAACAACTTT

Vigna unguiculata CGGACACAGTGAACGCGTTATTAGATTTAA

Phe162Ala TTAAATCTAATAACGCGTTCACTGTGTCCGAAAGTTGTTTTCTGAA | 926

TTT-GCT GCCAGCATCCCTAGCAAATCCATCATTCACAAGAAGTTGGGAAGA : AGTCACATCAAGCTGGCTGAACATGGTATC

ATGGATTTGCTAGGGAT

1142001 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene, oo

Plant & Targeted Altering Oligos - Alteration

ATCCCTAGCAAATCCAT alt Tolerance | GACACCTITGTTTAGTCAGTTGGATCIGACTGCTGCTCAGCTGCTT | §

P5CS GTGACGGACAACGACGCTAGAGATCCAAGTTTTAGAACACAACTA

Mesembryanthemum | ACTGAAACAGTGTATCAGTTGTTGGATCTAA crystallinum TTAGATCCAACAACTGATACACTGTTTCAGTTAGTTGTGTTCTAAA | 930

Phe125Ala ACTTGGATCTCTAGCGTCGTTGTCCGTCACAAGCAGCTGAGCAGC

TTT-GCT AGTCAGATCCAACTGACTAAACAAGGTGTC

ACAACGACGCTAGAGAT

ATCTCTAGCGTCGTTGT

® Salt Tolerance | GACACATTATTTAGCCAGCTGGATGTGACATCAGCTCAGCTTC 93

P5CS GTGACTGATAATGATGCTAGGGATGAAGCTTTCCGAAATCAACTTA

Vitis vinifera CTCAAACAGTGGATTCATTGTTAGCTTTGA

Phe130Ala TCAAAGCTAACAATGAATCCACTGTTTGAGTAAGTTGATTTCGGAA | 934

TIT-GCT AGCTTCATCCCTAGCATCATTATCAGTCACAAGAAGCTGAGCTGAT

GTCACATCCAGCTGGCTAAATAATGTGTC

ATAATGATGCTAGGGAT

ATCCCTAGCATCATTAT

Salt Tolerance GATACGCTGTTCACTCAGCTCGATGTGACATCGGCTCAGCTICTT | 937

P5CS GTGACGGATAACGATGCTCGAGATAAGGATTTCAGGAAGCAGCTT

Vigna aconitifolia ACTGAGACTGTGAAGTCGCTGTTGGCGCTGA

Phe129Ala TCAGCGCCAACAGCGACTTCACAGTCTCAGTAAGCTGCTTCCTGA | 938

TTT-GCT AATCCTTATCTCGAGCATCGTTATCCGTCACAAGAAGCTGAGCCG

ATGTCACATCGAGCTGAGTGAACAGCGTATC

ATAACGATGCTCGAGAT o ATCTCGAGCATCGTTAT 940 alt Tolerance AGAGATGTICTTAGTTCCAAAGAAATCTCACCTCTCACTTTCTCCG | 941

HKT1 TCTTCACAACAGTTGTCACGTTTGCAAACTGCGGATTTGTCCCCAC

Arabidopsis thaliana | GAATGAGAACATGATCATCTTTCGCAAAA

Ser207Val TTTTGCGAAAGATGATCATGTTCTCATTCGTGGGGACAAATCCGC | 942

TCC-GTC AGTTTGCAAACGTGACAACTGTTGTGAAGACGGAGAAAGTGAGAG

GTGAGATTTCTTTGGAACTAAGAACATCTCT

CAACAGTTGTCACGTTT

AAACGTGACAACTGTTG

Salt Tolerance CGAATGAGAACATGATCATCTTTCGCAAAAACTCTGGTCICAICTG | 945

HKT1 GCTCCTAATCCCTCTAGTACTGATGGGAAACACTTTGTTCCCTTGC : Arabidopsis thaliana | TTCTTGGTTTTGCTCATATGGGGACTTTA

GIn237Leu TAAAGTCCCCATATGAGCAAAACCAAGAAGCAAGGGAACAAAGTG | 946

CAA-CTA TTTCCCATCAGTACTAGAGGGATTAGGAGCCAGATGAGACCAGAG

TTTTTGCGAAAGATGATCATGTTCTCATTCG

1142/01 01:47 pm 03137.009 — [NY)793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration

AATCCCTCTAGTACTGA

TCAGTACTAGAGGGATT

Salt Tolerance AGTCTCTAGAAGGAATGAGTTCGTACGAGAA TGGATCGT | 949

HKT1 TGTTTCAAGTGGTGAGTTCGCGACACACCGGAGAAACTATAGTAG

Arabidopsis thaliana | ACCTCTCTACACTTTCCCCAGCTATCTTGGT

Asn332Ser ACCAAGATAGCTGGGGAAAGTGTAGAGAGGTCTACTATAGTTTCT | 950

AAT-AGT CCGGTGTGTCGCGAACTCACCACTTGAAACAACGATCCAACCAAC

TTCTCGTACGAACTCATTCCTTCTAGAGACT

AGTGGTGAGTTCGCGAC

® GTCGCGAACTCACCACT alt Tolerance | AGAGATGIGCTAAAGAAGAAAGGTCTCAAARATGGTGACCTTTICC | 9%

HKT1 GTCTTCACCACCGTGGTGACCTTTGCCAGTTGTGGGTTTGTCCCG

Eucalyptus ACCAATGAAAACATGATTATCTTCAGCAAAA camaldulensis TTTTGCTGAAGATAATCATGTTTTCATTGGTCGGGACAAACCCACA | 954

Ser256Val ACTGGCAAAGGTCACCACGGTGGTGAAGACGGAAAAGGTCACCA

TCG-GTG TTTTGAGACCTTTCTTCTTTAGCACATCTCT

CCACCGTGGTGACCTTT

AAAGGTCACCACGGTGG

Salt Tolerance CAATGAAAACATGATTATCTTCAGCAAAAACTCTGGCCTCCT 957

HKT1 GATTCTCATCCCTCIGGCCCTTCTTGGGAACATGCTGTTCCCATC

Eucalyptus GAGCCTACGTTTGACGCTTTGGCTCATCGG camaldulensis CCGATGAGCCAAAGCGTCAAACGTAGGCTCGATGGGAACAGCAT | 958

Gin286Leu GTTCCCAAGAAGGGCCAGAGGGATGAGAATCAGGAGGAGGCCA

CAG-CTG GAGTTTTTGCTGAAGATAATCATGTTTTCATTGG ® CATCCCTCIGGCCCTTC

GAAGGGCCAGAGGGATG | 960

Salt Tolerance AATCGTTGAATGGACTAAGCTCCTGTGAGAAAATCGTGGGCGCGC | 961

HKT1 TGTTTCAGTGCGTGAGCAGCAGACATACCGGCGAGACGGTCGTC

Eucalyptus GATCTGTCCACAGTTGCTCCCGCCATCTTGGT camaldulensis ACCAAGATGGCGGGAGCAACTGTGGACAGATCGACGACCGTCTC | 962

Asn381Ser GCCGGTATGTCTGCTGCTCACGCACTGAAACAGCGCGCCCACGA

AAC-AGC TTTTCTCACAGGAGCTTAGTCCATTCAACGATT

GTGCGTGAGCAGCAGAC

GTCTGCTGCTCACGCAC

1112/01 01:47 pm 03137.008 — [NY}793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Salt Tolerance AAAGCTCCACTGAAGAAGAAAGGGATCAACA ACTCTTCTCA : - HKT1 TTCTCGGTCACGGTCGICTCGTTTGCGAATGTGGGGCTCGTGCC

Oryza sativa GACAAATGAGAACATGGCAATCTTCTCCAAGA

Ser238Val TCTTGGAGAAGATTGCCATGTTCTCATTTGTCGGCACGAGCCCCA

TCC-GTC CATTCGCAAACGAGACGACCGTGACCGAGAATGAGAAGAGTGCA

ATGTTGATCCCTTTCTTCTTCAGTGGAGCTTT

TCACGGTCGICTCGTTT

AAACGAGACGACCGTGA

Call Tolerance | CAAATGAGAACATGGCAATCTTCTCCAAGAACCCGGGCCTCCTCC | 969

HKT1 TCCTGTTCATCGGCCIGATTCTTGCAGGCAATACACTTTACCCTCT ® Oryza sativa CTTCCTAAGGCTATTGATATGGTTCCTGGG

Gin268Leu CCCAGGAACCATATCAATAGCCTTAGGAAGAGAGGGTAAAGTGTA | 970

CAG-CTG TTGCCTGCAAGAATCAGGCCGATGAACAGGAGBAGGAGGCCCGG

GTTCTTGGAGAAGATTGCCATGTTCTCATTIG

CATCGGCCIGATTCTTG

CAAGAATCAGGCCGATG

HKT1 GTTCATGGCAGTGAGCGCAAGGCACTCGGGGGAGAACTCCATCG

Oryza sativa ACTGCTCACTCATCGCCCCTGCTGTTCTAGT

Asn363Ser ACTAGAACAGCAGGGGCGATGAGTGAGCAGTCGATGGAGTTCTC | 974

AAC-AGC CCCCGAGTGCCTTGCGCTCACTGCCATGAACAATGCATTGATAAT

CTTCTGGTAAGAGCTGAGTCCATCAAAGACTG :

GGCAGTGAGCGCAAGGC

GCCTTGCGCTCACTGCC

®

HKT1 TATCAGTCACCGTTGICTCCTGTGCGAATGCAGGACTCGTGCCCA

Triticum aestivum CAAATGAGAACATGGTCATCTTCTCAAAGAA

Ala240Val TTCTTTGAGAAGATGACCATGTTCTCATTTGTGGGCACGAGTCCT | 978

GCC-GTC GCATTCGCACAGGAGACAACGGTGACTGATAGTGAGAAGAGCAC

GATGTTGATCCCTTTCTTGTTCAGTGGGGCAC

CACCGTTGICTCCTGTG

CACAGGAGACAACGGTG

: HKT1 GCTGCTGAGTGGCCIGATGCTCGCAGGCAATACATTGTTCCCTCT

Triticum aestivum | CTTCCTGAGGCTACTGGTGTGGTTCCTGGG

GIn270Leu CCCAGGAACCACACCAGTAGCCTCAGGAAGAGAGGGAACAATGT | 982 + 25 CAG-CTG ATTGCCTGCGAGCATCAGGCCACTCAGCAGCAACAAGAGGCCTG

AATTCTTTGAGAAGATGACCATGTTCTCATTTG

GAGTGGCCTIGATGCTCG

1112101 01:47 pm 03137.009 — [NY}793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration

CGAGCATCAGGCCACIC

: Salt Tolerance CAGTCTTTGATGGGCTCAGCTCTTATCAGAAGACTGTCAATGCAT 985

HKT1 CTTCATGGTGGTGAGTGCGAGGCACTCAGGGGAGAATTCCATCG

Triticum aestivum ACTGCTCGCTCATGTCCCCTGCCATTATAGT

Asn365Ser ACTATAATGGCAGGGGACATGAGCGAGCAGTCGATGGAATTCTCC| 986

AAT-AGT CCTGAGTGCCTCGCACTCACCACCATGAAGAATGCATTGACAGTC

TTCTGATAAGAGCTGAGCCCATCAAAGACTG

GGTGGTGAGTGCGAGGC

GCCTCGCACTCACCACC

® Freezing Tolerance TTTTTTTTGTTTTCGTTTTCAAAAACAAAATCTTTGAATTTTATGGCA | 989 proline oxidase ACCCGTCTTCTCTGAACAAACTTTATCCGGCGATCTTACCGTTTAC precursor CCGCTTTTAGCCCGGTGGGTCCTCCCA

Arabidopsis thaliana | TGGGAGGACCCACCGGGCTAAAAGCGGGTAAACGGTAAGATCGC

Arg7Term CGGATAAAGTTTGTTCAGAGAAGACGGGTTGCCATAAAATTCAAA

CGA-TGA GATTTTGTTTTTGAAAACGAAAACAAAAAAAA

GTCTTCTCTGAACAAAC

GTTTGTTCAGAGAAGAC

Freezing Tolerance | TCAAAAACAAAATCTTTGAATTTTATGGCAACCCGTCTTCTCAGAA 993 proline oxidase CAAACTTTATCCGGTGATCTTACCGTTTACCCGCTTTTAGCCCGGT precursor GGGTCCTCCCACCGTGACTGCTTCCACCG

Arabidopsis thaliana | CGGTGGAAGCAGTCACGGTGGGAGGACCCACCGGGCTAAAAGC 994

Arg13Term GGGTAAACGGTAAGATCACCGGATAAAGTTTGTTCTGAGAAGACG

CGA-TGA GGTTGCCATAAAATTCAAAGATTTTGTTTTTGA

TTATCCGGIGATCTTAC | 995 ® GTAAGATCACCGGATAA 996

Freezing Tolerance | AAAATCTTTGAATTTTATGGCAACCCGTCTTCTCCGAACAAACTTT 997 proline oxidase ATCCGGCGATCTTAGCGTTTACCCGCTTTTAGCCCGGTGGGTCCT precursor CCCACCGTGACTGCTTCCACCGCCGTCGTC

Arabidopsis thaliana | GACGACGGCGGTGGAAGCAGTCACGGTGGGAGGACCCACCGGG | 998

Tyri5Term CTAAAAGCGGGTAAACGCTAAGATCGCCGGATAAAGTTTGTTCGG

TAC-TAG AGAAGACGGGTTGCCATAAAATTCAAAGATTTT

CGATCTTAGCGTTTACC | 999

GGTAAACGCTAAGATCG 1000 11112001 01:47pm 03137.009 — [NY1793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Freezing Tolerance CTTTGAATTTTATGGCAACCCGTCTTCTCCGAACAAACTTTATCCG | 1001 proline oxidase GCGATCTTACCGTTAACCCGCTTTTAGCCCGGTGGGTCCTCCCAC precursor CGTGACTGCTTCCACCGCCGTCGTCCCGGA

Arabidopsis thaliana | TCCGGGACGACGGCGGTGGAAGCAGTCACGGTGGGAGGACCCA | 1002

Leu17Term CCGGGCTAAAAGCGGGTTAACGGTAAGATCGCCGGATAAAGTTT

TTA-TAA GTTCGGAGAAGACGGGTTGCCATAAAATTCAAAG

TTACCGTTAACCCGCTT

AAGCGGGTTAACGGTAA 1004

Freezing Tolerance | CCGGTGGGTCCTCCCACCGTGACTGCTTCCACCGCCGTCGTCCC | 1005 9 proline oxidase GGAGATTCTCTCCTTTIGACAACAAGCACCGGAACCACCTCTTCA precursor CCACCCAAAACCCACCGAGCAATCTCACGATG

Arabidopsis thaliana | CATCGTGAGATTGCTCGGTGGGTTTTGGGTGGTGAAGAGGTGGT | 1006

Gly4d2Term TCCGGTGCTTGTTGTCAAAAGGAGAGAATCTCCGGGACGACGGC

GGA-TGA GGTGGAAGCAGTCACGGTGGGAGGACCCACCGE

TCTCCTTTIGACAACAA 1007

TTGTTGTCAAAAGGAGA 1008 ead Tolerance ACATGAAGCA AAA ATTGAA ATTA T1009 cyclic nucleotide- AAACTATGAATTTCTIGACAAGAGAAGTTTGTAAGGTCAGTGTTCCA regulated ion channel | GATTTGTCTCATTGAATTCTAAGTCGTGA

Arabidopsis thaliana | TCACGACTTAGAATTCAATGAGACAAATCTGGAACACTGACCTTAC | 1010

ArgdTerm AAACTTCTCTTGTCAGAAATTCATAGTTTGAGACTAATAAGATTCAA

CGA-TGA TACAAACAGAGATTTCACTGCTTCATGT

TGAATTTCTIGACAAGAG 1011

PY CTCTTGTCAGAAATTCA 1012

Lead Tolerance TGAAGCAGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCAAA | 1013 cyclic nucleotide- CTATGAATTTCCGATAAGAGAAGTTTGTAAGGTCAGTGTTCCAGAT regulated ion channel | TTGTCTCATTGAATTCTAAGTCGTGAAGC

Arabidopsis thaliana | GCTTCACGACTTAGAATTCAATGAGACAAATCTGGAACACTGACCT | 1014

GIn5Term TACAAACTTCTCTTATCGGAAATTCATAGTTTGAGACTAATAAGATT

CAA-TAA CAATACAAACAGAGATTTCACTGCTTCA

ATTTCCGATAAGAGAAG

CTTCTCTTATCGGAAAT 1016 © 25 Lead Tolerance AGCAGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCAAACTAT | 1017 cyclic nucleotide- GAATTTCCGACAATAGAAGTTTGTAAGGTCAGTGTTCCAGATTTGT regulated ion channel | CTCATTGAATTCTAAGTCGTGAAGCTTA

Arabidopsis thaliana | TAAGCTTCACGACTTAGAATTCAATGAGACAAATCTGGAACACTGA | 1018

Glu6Term CCTTACAAACTTCTATTGTCGGAAATTCATAGTTTGAGACTAATAA

GAG-TAG GATTCAATACAAACAGAGATTTCACTGCT 1112/01 01:47 pm 03137.009 — {NY]783559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene, }

Plant & Targeted Altering Oligos

Alteration

TCCGACAATAGAAGTTT 1019

AAACTTCTATTGTCGGA 1020

Lead Tolerance AGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCAAACTATGAA | 1021 cyclic nucleotide- TTTCCGACAAGAGTAGTTTGTAAGGTCAGTGTTCCAGATTTGTCTC regulated jon channel | ATTGAATTCTAAGTCGTGAAGCTTAATT

Arabidopsis thaliana | AATTAAGCTTCACGACTTAGAATTCAATGAGACAAATCTGGAACAC | 1022

Lys7Term TGACCTTACAAACTACTCTTGTCGGAAATTCATAGTTTGAGACTAA

AAG-TAG TAAGATTCAATACAAACAGAGATTTCACT

GACAAGAGTAGTTTGTA 1023 ® TACAAACTACTCTTGTC 1024

Lead Tolerance CATTGAATTCTAAGTCGTGAAGCTTAATTCGATTCTTCTTCACTITC | 1025 cyclic nucleotide- TCGGATCAGGTTTIAAGATTGGAAGTCGGATAAGACTTCCTCCGA regulated ion channel | CGTGGAATATTCCGGTAAAAACGAGATTC

Arabidopsis thaliana | GAATCTCGTTTTTACCGGAATATTCCACGTCGGAGGAAGTCTTATC | 1026

GIn12Term CGACTTCCAATCTTAAAACCTGATCCGAGAAAGTGAAGAAGAATC

CAA-TAA GAATTAAGCTTCACGACTTAGAATTCAATG

TCAGGTTTTAAGATTGG 1027

CCAATCTTAAAACCTGA 1028 cyclic nucleotide- GTTATGAATCACCGCTAAGACGAGTTTGTGAGGTTTCAGGATTGG gated calmodulin- AAATCAGAGAGAAGCTCTGAGGGAAATTTTC binding ion channel | GAAAATTTCCCTCAGAGCTTCTCTCTGATTTCCAATCCTGAAACCT | 1030 (CBP4) CACAAACTCGTCTTAGCGGTGATTCATAACTTTAGCCAATGCATCA ® Nicotiana Tabacum | ACCTGCTCAACGTGGGGGATTGACTTCCA

GIn5Term ATCACCGCTAAGACGAG 2 CAA-TAA CTCGTCTTAGCGGTGAT 1032 cyclic nucleotide AATCACCGCCAAGACTAGTTTGTGAGGTTTCAGGATTGGAAATCA gated calmodulin- GAGAGAAGCTCTGAGGGAAATTTTCATGCTA : binding ion channel | TAGCATGAAAATTTCCCTCAGAGCTTCTCTCTGATTTCCAATCCTG | 1034 (CBP4) AAACCTCACAAACTAGTCTTGGCGGTGATTCATAACTTTAGCCAAT

Nicotiana Tabacum | GCATCAACCTGCTCAACGTGGGGGATTGA

Gly7Term GCCAAGACTAGTTTGTG 1035

GAG-TAG CACAAAGTAGTCTIGGC 11/12/01 01:47 pm 03137.009 ~ [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Lead Tolerance GAGCAGGTTGATGCATTGGCTAAAGTTATGAATCACCGCCAAGAC | 1037 cyclic nucleotide- GAGTTTGTGAGGTTTTAGGATTGGAAATCAGAGAGAAGCTCTGAG gated calmodulin- GGAAATTTTCATGCTAAAGGTGGAGTCCACC binding ion channel | GGTGGACTCCACCTTTAGCATGAAAATTTCCCTCAGAGCTTCTCTC| 1038 (CBP4) TGATTTCCAATCCTAAAACCTCACAAACTCGTCTTGGCGGTGATTC

Nicotiana Tabacum | ATAACTTTAGCCAATGCATCAACCTGCTC

Gln12Term TGAGGTTTTAGGATTGG 1039

CAG-TAG

CCAATCCTAAAACCTCA 1040

Lead Tolerance TGATGCATTGGCTAAAGTTATGAATCACCGCCAAGACGAGTTTGT [| 1041 ® cyclic nucleotide- GAGGTTTCAGGATTGTAAATCAGAGAGAAGCTCTGAGGGAAATTT gated calmodulin- TCATGCTAAAGGTGGAGTCCACCGAAGTAAA binding ion channel | TTTACTTCGGTGGACTCCACCTTTAGCATGAAAATTTCCCTCAGAG | 1042 (CBP4) CTTCTCTCTGATTTACAATCCTGAAACCTCACAAACTCGTCTTGGC

Nicotiana Tabacum | GGTGATTCATAACTTTAGCCAATGCATCA

Trp14Term CAGGATTGTAAATCAGA 1043

TGG-TGA TCTGATTTACAATOCTG 1044 cyclic nucleotide- AGGTTTCAGGATTGGTAATCAGAGAGAAGCTCTGAGGGAAATTTT gated calmodulin- CATGCTAAAGGTGGAGTCCACCGAAGTAAAG binding ion channel | CTTTACTTCGGTGGACTCCACCTTTAGCATGAAAATTTCCCTCAGA | 1046 (CBP4) GCTTCTCTCTGATTACCAATCCTGAAACCTCACAAACTCGTCTTGG

Nicotiana Tabacum | CGGTGATTCATAACTTTAGCCAATGCATC

Lys15Term AGGATTGGTAATCAGAG 1047 ® AAA-TAA CTCTGATTACCAATCCT calmodulin binding TGGAGATAATGATGTAAAGAGAGGACAGATATGTTAGATTTCAGG transport protein ACTGCAAATCAGAGCAATCTGTTATCTCAG

Hordeum vulgare CTGAGATAACAGATTGCTCTGATTTGCAGTCCTGAAATCTAACATA | 1050

Glu2Term TCTGTCCTCTCTTTACATCATTATCTCCACCAGGCGAACAGTTAGC

GAA-TAA AGCTAAGAGTGGTAGATCAATTCTTCAAG

TAATGATGTAAAGAGAG 1051

CTCTCTTTACATCATTA 1052 1142/08 01:47 pm 03137.009 — [NY]793559.4

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Lead Tolerance GAAGAATTGATCTACCACTCTTAGCTGCTAACTGTTCGCCTGGTG | 1053 calmodulin binding | GAGATAATGATGGAATGAGAGGACAGATATGTTAGATTTCAGGAC transport protein TGCAAATCAGAGCAATCTGTTATCTCAGAGA

Hordeum vulgare TCTCTGAGATAACAGATTGCTCTGATTTGCAGTCCTGAAATCTAAC | 1054

Arg3Term ATATCTGTCCTCTCATTCCATCATTATCTCCACCAGGCGAACAGTT

AGA-TGA AGCAGCTAAGAGTGGTAGATCAATTCTTC

TGATGGAATGAGAGGAC

GTCCTCTCATTCCATCA

Lead Tolerance GAATTGATCTACCACTCTTAGCTGCTAACTGTTCGCCTGGTGGAG | 1057 ® camodulin binding | ATAATGATGGAAAGATAGGACAGATATGTTAGATTTCAGGACTGC transport protein AAATCAGAGCAATCTGTTATCTCAGAGAACG

Hordeum vulgare CGTTCTCTGAGATAACAGATTGCTCTGATTTGCAGTCCTGAAATCT | 1058

GludTerm AACATATCTGTCCTATCTTTCCATCATTATCTCCACCAGGCGAACA

GAG-TAG GTTAGCAGCTAAGAGTGGTAGATCAATTC

TGGAAAGATAGGACAGA 1059

TCTGTCCTATCTTTCCA | 1060

Lead Tolerance | ATCTACCACTCTTAGCTGCTAACTGTTCGCCTGGTGGAGATAATG | 1061 calmodulin binding | ATGGAAAGAGAGGACTGATATGTTAGATTTCAGGACTGCAAATCA transport protein GAGCAATCTGTTATCTCAGAGAACGCAGTTT

Hordeum vulgare AAACTGCGTTCTCTGAGATAACAGATTGCTCTGATTIGCAGTCCTG | 1062

Arg6Term AAATCTAACATATCAGTCCTCTCTTTCCATCATTATCTCCACCAGG

AGA-TGA CGAACAGTTAGCAGCTAAGAGTGGTAGAT

GAGAGGACTGATATGTT 1063

PY AACATATCAGTCCTCTC 1064

Lead Tolerance CCACTCTTAGCTGCTAACTGTTCGCCTGGTGGAGATAATGATGGA | 1065 calmodulin binding | AAGAGAGGACAGATAGGTTAGATTTCAGGACTGCAAATCAGAGCA transport protein ATCTGTTATCTCAGAGAACGCAGTTTCACCA

Hordeum vulgare TGGTGAAACTGCGTTCTCTGAGATAACAGATTGCTCTGATTTGCA | 1066

Tyr7Term GTCCTGAAATCTAACCTATCTGTCCTCTCTTTCCATCATTATCTCCA

TAT-TAG CCAGGCGAACAGTTAGCAGCTAAGAGTGG

GACAGATAGGTTAGATT 1067

AATCTAACCTATCTGTC 1068 2 4-DB resistance ATCCTTCTCTGAGAAAAAACAACAGATCCGAATTTTATCTTTAATCA | 1069 3-ketoacyl-CoA GCCGGAAAAAATGTAGAAAGCGATCGAGAGACAACGCGTTCTTCT : thiolase TGAGCATCTCCGACCTTCTTCTTCTTCTT

Arabidopsis thaliana | AAGAAGAAGAAGAAGGTCGGAGATGCTCAAGAAGAACGCGTTGT | 1070

Glu2Term CTCTCGATCGCTTTCTACATTTTTTCCGGCTGATTAAAGATAAAATT

GAG-TAG CGGATCTGTTGTTITTTCTCAGAGAAGGAT 1112001 01:47 pm 03137.009 — [NY)793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

AAAAAATGTAGAAAGCG 1071 ‘ CGCTTTCTACATTTTIT 1072

JA -DB resistance ACSA KAA AACAACAGA AA A AA A () 3-ketoacyl-CoA CGGAAAAAATGGAGTAAGCGATCGAGAGACAACGCGTTCTTCTTG thiolase AGCATCTCCGACCTTCTTCTTCTTCTTCGC

Arabidopsis thaliana | GCGAAGAAGAAGAAGAAGGTCGGAGATGCTCAAGAAGAACGCGT | 1074

Lys3Term TGTCTCTCGATCGCTTACTCCATTTTTTCCGGCTGATTAAAGATAA

AAA-TAA AATTCGGATCTGTTGTTTTTTCTCAGAGAAG

AAATGGAGTAAGCGATC 1075

C GATCGCTTACTCCATTT

24-DBresistance | GAAAAAACAACAGATCCGAATTTTATCTTTAATCAGCCGGAAAAAA | 1077 3-ketoacyl-CoA TGGAGAAAGCGATCTAGAGACAACGCGTTCTTCTTGAGCATCTCC thiolase GACCTTCTTCTTCTTCTTCGCACAATTACG

Arabidopsis thaliana | CGTAATTGTGCGAAGAAGAAGAAGAAGGTCGGAGATGCTCAAGA | 1078

GlusTerm AGAACGCGTTGTCTCTAGATCGCTTTCTCCATTTTTTCCGGCTGAT

GAG-TAG TAAAGATAAAATTCGGATCTGITGTTTTTTC

AAGCGATCTAGAGACAA

TTGTCTCTAGATCGCTT 1080 2,4-DB resistance AAAACAACAGATCCGAATTTTATCTTTAATCAGCCGGAAAAAATGG | 1081 3-ketoacyl-CoA AGAAAGCGATCGAGIGACAACGCGTTCTTCTTGAGCATCTCCGAC thiolase CTTCTTCTTCTTCTTCGCACAATTACGAGG

Arabidopsis thaliana | CCTCGTAATTGTGCGAAGAAGAAGAAGAAGGTCGGAGATGCTCAA| 1082

Arg7Term GAAGAACGCGTTGTCACTCGATCGCTTTCTCCATTTTTTCCGGCT

AGA-TGA GATTAAAGATAAAATTCGGATCTGTTGTTTT ® CGATCGAGTGACAACGC

GCGTTGTCACTCGATCG 1084 2,4-DB resistance ACAACAGATCCGAATTTTATCTTTAATCAGCCGGAAAAAATGGAGA | 1085 3-ketoacyl-CoA AAGCGATCGAGAGATAACGCGTTCTTCTTGAGCATCTCCGACCTT thiolase CTTCTTCTTCTTCGCACAATTACGAGGCTT

Arabidopsis thaliana | AAGCCTCGTAATTGTGCGAAGAAGAAGAAGAAGGTCGGAGATGC | 1086

GIn8Term TCAAGAAGAACGCGTTATCTCTCGATCGCTTTCTCCATTTTTTCCG

CAA-TAA GCTGATTAAAGATAAAATTCGGATCTGTTGT

TCGAGAGATAACGCGTT 1087

AACGCGTTATCTCTCGA 1088 1412/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration 2,4-DB resistance GAGAGACAAAGAGTTCTTCTTGAACATCTCCGTCCTTCTTCTTCTT | 1089 : glyoxysomal beta- CCTCTCACAGCTTTTAAGGCTCTCTCTCTGCTTCAGCTTGCTTGGC ketoacyol-thiolase TGGCGGACAGTGCTGCGTATCAGAGGACCT precursor AGGTCCTCTGATACGCAGCACTGTCCCCAGCCAAGCAAGCTGAA | 1090

Brassica napus GCAGAGAGAGAGCCTTAAAAGCTGTGAGAGGAAGAAGAAGAAGG

Glu26Term ACGGAGATGTTCAAGAAGAACTCTTTGTCTCTC

GAA-TAA ACAGCTTTTAAGGCTCT 1091

AGAGCCTTAAAAGCTGT 1092

C [2,4-DB resistance TTGAACATCTCCGTCCTTCTTCTTCTTCCTCTCACAGCTTTGAAGG | 1093 glyoxysomal beta- CTCTCTCTCTGCTTGAGCTTGCTTGGCTGGGGACAGTGCTGCGTA ketoacyol-thiolase TCAGAGGACCTCTCTCTATGGAGATGATGT precursor ACATCATCTCCATAGAGAGAGGTCCTCTGATACGCAGCACTGTCC

Brassica napus CCAGCCAAGCAAGCTCAAGCAGAGAGAGAGCCTTCAAAGCTGTG

Ser32Term AGAGGAAGAAGAAGAAGGACGGAGATGTTCAA

TCA-TGA CTCTGCTTGAGCTTGCT 1095

AGCAAGCTCAAGCAGAG 1096 i glyoxysomal beta- TCTGCTTCAGCTTGATTGGCTGGGGACAGTGCTGCGTATCAGAG ketoacyol-thiolase GACCTCTCTCTATGGAGATGATGTAGTCATT precursor AATGACTACATCATCTCCATAGAGAGAGGTCCTCTGATACGCAGC | 1098

Brassica napus ACTGTCCCCAGCCAATCAAGCTGAAGCAGAGAGAGAGCCTTCAAA

Cys34Term GCTGTGAGAGGAAGAAGAAGAAGGACGGAGA

TGC-TGA TCAGCTTGATTGGCTGG 1099 ® CCAGCCAATCAAGCTGA 1100 glyoxysomal beta- TGCTTCAGCTTGCTAGGCTGGGGACAGTGCTGCGTATCAGAGGA ketoacyol-thiolase CCTCTCTCTATGGAGATGATGTAGTCATTGT precursor ACAATGACTACATCATCTCCATAGAGAGAGGTCCTCTGATACGCA | 1102

Brassica napus GCACTGTCCCCAGCCTAGCAAGCTGAAGCAGAGAGAGAGCCTTC

Leu35Term AAAGCTGTGAGAGGAAGAAGAAGAAGGACGGA

TTG-TAG AGCTTGCTAGGCTGGGG 1103

CCCCAGCCTAGCAAGCT 1104

M201 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : 2,4-DB resistance TCACAGCTTTGAAGGCTCTCTCTCTGCTTCAGCTTGCTTGGCTGG | 1105 glyoxysomal beta- GGACAGTGCTGCGTAGCAGAGGACCTCTCTCTATGGAGATGATGT ketoacyol-thiolase AGTCATTGTTGCGGCACATAGGACTGCACTA precursor TAGTGCAGTCCTATGTGCCGCAACAATGACTACATCATCTCCATA | 1106

Brassica napus GAGAGAGGTCCTCTGCTACGCAGCACTGTCCCCAGCCAAGCAAG

Tyr42Term CTGAAGCAGAGAGAGAGCCTTCAAAGCTGTGA

TAT-TAG GCTGCGTAGCAGAGGAC 1107

GTCCTCTGCTACGCAGC 1108 2,4-DB resistance CAACAGACAGCAAGTGTTGCTCCAGCATCTCCGCCCTTCTAATTC | 1109 ® 3-ketoacyl-CoA TTCTTCTCACAATTAGGAGTCCGCTCTTGCCGCATCAGTATGTGCT thiolase B GCAGGGGATAGCGCCGCATATCATAGGGCT

Mangifera indica AGCCCTATGATATGCGGCGCTATCCCCTGCAGCACATACTGATGC | 1110

Tyr25Term GGCAAGAGCGGACTCCTAATTGTGAGAAGAAGAATTAGAAGGGC

TAC-TAG GGAGATGCTGGAGCAACACTTGCTGTCTGTTG

CACAATTAGGAGTCCGC

GCGGACTCCTAATTGTG 1112 2 4-DB resistance AACAGACAGCAAGTGTTGCTCCAGCATCTCCGCCCTTCTAATICTT | 1113 3-ketoacyol-CoA CTTCTCACAATTACTAGTCCGCTCTTGCCGCATCAGTATGTGCTGC thiolase B AGGGGATAGCGCCGCATATCATAGGGCTT

Magnifera indica AAGCCCTATGATATGCGGCGCTATCCCCTGCAGCACATACTGATG | 1114

Glu26Term CGGCAAGAGCGGACTAGTAATTGTGAGAAGAAGAATTAGAAGGG

GAG-TAG CGGAGATGCTGGAGCAACACTTGCTGTCTGTT

ACAATTACTAGTCCGCT 1115 ® AGCGGACTAGTAATTGT 1116 3-ketoacy\ol-CoA CGCTCTTGCCGCATGAGTATGTGCTGCAGGGGATAGCGCCGCAT thiolase B ATCATAGGGCTTCTGTTTATGGAGACGATGT

Mangifera indica ACATCGTCTCCATAAACAGAAGCCCTATGATATGCGGCGCTATCC | 1118

Ser32Term CCTGCAGCACATACTCATGCGGCAAGAGCGGACTCGTAATTGTGA

TCA-TGA GAAGAAGAATTAGAAGGGCGGAGATGCTGGA

TGCCGCATGAGTATGTG 1119

CACATACTCATGCGGCA 1120 14/12/01 01:47 pm 03137.008 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos 2,4-DB resistance TCTCCGCCCTTCTAATTCTTCTTCTCACAATTACGAGTCCGCTCTT | 1121 3-ketoacyl-CoA GCCGCATCAGTATGAGCTGCAGGGGATAGCGCCGCATATCATAG thiolase B GGCTTCTGTTITATGGAGACGATGTGGTGATT

Mangifera indica AATCACCACATCGTCTCCATAAACAGAAGCCCTATGATATGCGGC | 1122

Cys34Term GCTATCCCCTGCAGCTCATACTGATGCGGCAAGAGCGGACTCGT

TGT-TGA AATTGTGAGAAGAAGAATTAGAAGGGCGGAGA

TCAGTATGAGCTGCAGG 1123

CCTGCAGCTCATACTGA 1124 2,4-DB resistance TCACAATTACGAGTCCGCTCTTGCCGCATCAGTATGTGCTGCAGG | 1125 ® 3-ketoacyl-CoA GGATAGCGCCGCATAGCATAGGGCTTCTGTTTATGGAGACGATGT thiolase B GGTGATTGTGGCAGCTCATCGTACTGCACTT

Mangifera indica AAGTGCAGTACGATGAGCTGCCACAATCACCACATCGTCTCCATA | 1126

Tyrd2Term AACAGAAGCCCTATGCTATGCGGCGCTATCCCCTGCAGCACATAC

TAT-TAG TGATGCGGCAAGAGCGGACTCGTAATTGTGA

GCCGCATAGCATAGGGC 1127

GCCCTATGCTATGCGGC 1128 2,4-DB resistance GAAGGCGATCAACAGGCAGAGCATTTTGCTACATCATCTCCGGCC | 1129 3-ketoacyl-CoA TTCTTCTTCCGCTTAGACAAATGAATCTTCGCTCTCTGCATCGGTT thiolase TGTGCAGCTGGGGATAGTGCTTCGTATCAA

Cucumis sativus TTGATACGAAGCACTATCCCCAGCTGCACAAACCGATGCAGAGAG | 1130

Tyr22Tem CGAAGATTCATTTGTCTAAGCGGAAGAAGAAGGCCGGAGATGATG

TAC-TAG TAGCAAAATGCTCTGCCTGTTGATCGCCTTC

TCCGCTTAGACAAATGA 1131 ® TCATTTGTCTAAGCGGA 1132 3-ketoacyl-CoA CCGCTTACACAAATTAATCTTCGCTCTCTGCATCGGTTTGTGCAGC thiolase TGGGGATAGTGCTTCGTATCAAAGGACAT

Cucumis sativus ATGTCCTTTGATACGAAGCACTATCCCCAGCTGCACAAACCGATG | 1134

Glu25Term CAGAGAGCGAAGATTAATTTGTGTAAGCGGAAGAAGAAGGCCGG

GAA-TAA AGATGATGTAGCAAAATGCTCTGCCTGTTGAT

ACACAAATTAATCTTCG 1135

CGAAGATTAATTTGTGT 1136 11112001 01:47 pm 03137.009 — NY1793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration 2,4-DB resistance GGCAGAGCATTTTGCTACATCATCTCCGGCCTTCTTCTTCCGCTTA | 1137 3-ketoacyl-CoA CACAAATGAATCTTAGCTCTCTGCATCGGTTTGTGCAGCTGGGGA thiolase TAGTGCTTCGTATCAAAGGACATCGGTGTT

Cucumis sativus AACACCGATGTCCTTTGATACGAAGCACTATCCCCAGCTGCACAA | 1138

Ser27Term ACCGATGCAGAGAGCTAAGATTCATTTGTGTAAGCGGAAGAAGAA

TCG-TAG GGCCGGAGATGATGTAGCAAAATGCTCTGCC

TGAATCTTAGCTCTCTG 1139

CAGAGAGCTAAGATTCA 1140 24-DB rosstance | TGCTACATCATCTCCGGGCTTCTTCTTCCGCTTACACARATGAATC @® 3-ketoacyl-CoA TTCGCTCTCTGCATAGGTTTGTGCAGCTGGGGATAGTGCTTCGTA thiolase TCAAAGGACATCGGTGTTTGGAGATGATGT

Cucumis sativus ACATCATCTCCAAACACCGATGTCCTTTGATACGAAGCACTATCCC | 1142

Ser31Term CAGCTGCACAAACCTATGCAGAGAGCGAAGATTCATTTGTGTAAG

TCG-TAG CGGAAGAAGAAGGCCGGAGATGATGTAGCA

CTCTGCATAGGTTTGTG 1143

CACAAACCTATGCAGAG

3-ketoacyl-CoA TCTGCATCGGTTTGAGCAGCTGGGGATAGTGCTTCGTATCAAAGG thiolase ACATCGGTGTTTGGAGATGATGTCGTGATT

Cucumis sativus AATCACGACATCATCTCCAAACACCGATGTCCTTTGATACGAAGCA | 1146

Cys33Term CTATCCCCAGCTGCTCAAACCGATGCAGAGAGCGAAGATTCATTT

TGT-TGA GTGTAAGCGGAAGAAGAAGGCCGGAGATGA

TCGGTTTGAGCAGCTGG 1147 ® CCAGCTGCTCAAACCGA 1148 3-ketoacyl-CoA TTCATCTTCGGCTTAGAGCCATGAATCTTCGCTCTCTGCATCGGTT thiolase TGTGCAGCTGGGGATAGTGCGTCGTATCAA

Cucurbita sp. TTGATACGACGCACTATCCCCAGCTGCACAAACCGATGCAGAGAG

Tyr22Term CGAAGATTCATGGCTCTAAGCCGAAGATGAAGGCCGGAGATGAT

TAT-TAG GTAGCAGAATGCTCTGCCTGTTGATTGCCTTC

TCGGCTTAGAGCCATGA 1151

TCATGGCTCTAAGCCGA 1152 1412/01 01:47 pm 03137.063 — [NY]793553.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos 2,4-DB resistance ATCAACAGGCAGAGCATTCTGCTACATCATCTCCGGCCTTCATCTT | 1153 } 3-ketoacyl-CoA CGGCTTATAGCCATTAATCTTCGCTCTCTGCATCGGTTTGTGCAGC thiolase TGGGGATAGTGCGTCGTATCAAAGAACGT

Cucurbita sp. ACGTTCTTTGATACGACGCACTATCCCCAGCTGCACAAACCGATG | 1154

Glu25Term CAGAGAGCGAAGATTAATGGCTATAAGCCGAAGATGAAGGCCGG

GAA-TAA AGATGATGTAGCAGAATGCTCTGCCTGTTGAT

ATAGCCATTAATCTTCG 1165

CGAAGATTAATGGCTAT 1156 2,4-DB resistance GGCAGAGCATTCTGCTACATCATCTCCGGCCTTCATCTTCGGCTT | 1157 ® 3-ketoacyl-CoA ATAGCCATGAATCTTAGCTCTCTGCATCGGTTTGTGCAGCTGGGG : thiolase ATAGTGCGTCGTATCAAAGAACGTCGGTGTT

Cucurbita sp. AACACCGACGTTCTTTGATACGACGCACTATCCCCAGCTGCACAA | 1158

Ser27Term ACCGATGCAGAGAGCTAAGATTCATGGCTATAAGCCGAAGATGAA

TCG-TAG GGCCGGAGATGATGTAGCAGAATGCTCTGCC

TGAATCTTAGCTCTCTG 1159

CAGAGAGCTAAGATTCA 1160 2,4-DB resistance TGCTACATCATCTCCGGCCTTCATCTTCGGCTTATAGCCATGAATC | 1161 3-ketoacyl-CoA TTCGCTCTCTGCATAGGTTTGTGCAGCTGGGGATAGTGCGTCGTA thiolase TCAAAGAACGTCGGTGTTTGGAGATGATGT

Cucurbita sp. ACATCATCTCCAAACACCGACGTTCTTTGATACGACGCACTATCCC | 1162

Ser31Term CAGCTGCACAAACCTATGCAGAGAGCGAAGATTCATGGCTATAAG

TCG-TAG CCGAAGATGAAGGCCGGAGATGATGTAGCA

CTCTGCATAGGTTTGTG 1163 ® CACAAACCTATGCAGAG 1164 3-ketoacyl-CoA TCTGCATCGGTTTGAGCAGCTGGGGATAGTGCGTCGTATCAAAGA thiolase ACGTCGGTGTTTGGAGATGATGTCGTGATA

Cucurbita sp. TATCACGACATCATCTCCAAACACCGACGTTCTTTGATACGACGCA | 1166

Cys33Term CTATCCCCAGCTGCTCAAACCGATGCAGAGAGCGAAGATTCATGG

TGT-TGA CTATAAGCCGAAGATGAAGGCCGGAGATGA

TCGGTTTGAGCAGCTGG 1167

CCAGCTGCTCAAACCGA 1168 4 DB resistance ATA A AZ ) A 3

Pex14 CTATGGCAACTCATTAGCAAACGCAACCTCCTTCCGATTTTCCCGC

Arabidopsis thaliana | TCTTGCCGATGAAAATTCCCAGATTCCAG

GIn5Term CTGGAATCTGGGAATTTTCATCGGCAAGAGCGGGAAAATCGGAA 1170

CAG-TAG GGAGGTTGCGTTTGCTAATGAGTTGCCATAGCAGCTCACTAACCT

TGGAAGAATCCAAGCGGCAAAAGAGACTATGA

111201 01:47 pm 03137.009 — {NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

CAACTCATIAGCAAACG 1171

CGTTTGCTAATGAGTTG 1172 2.4 DB resistance TAGTCTCTTTTGCCGCTTGGATTCTTCCAAGGTTAGTGAGCTGCTA | 1173

Pex14 TGGCAACTCATCAGTAAACGCAACCTCCTTCCGATTTTCCCGCTCT

Arabidopsis thaliana | TGCCGATGAAAATTCCCAGATTCCAGGTT

Gin6Term AACCTGGAATCTGGGAATTTTCATCGGCAAGAGCGGGAAAATCGG | 1174

CAA-TAA AAGGAGGTTGCGTTTACTGATGAGTTGCCATAGCAGCTCACTAAC

CTTGGAAGAATCCAAGCGGCAAAAGAGACTA

CTCATCAGTAAACGCAA 1175 ® TTGCGTTTACTGATGAG 1176 2,4 DB resistance CTTTTGCCGCTTGGATTCTTCCAAGGTTAGTGAGCTGCTATGGCA | 1177

Pex14 ACTCATCAGCAAACGTAACCTCCTTCCGATTTTCCCGCTCTTGCCG

Arabidopsis thaliana | ATGAAAATTCCCAGATTCCAGGTTCAATTT

Gin8Term AAATTGAACCTGGAATCTGGGAATTTTCATCGGCAAGAGCGGGAA | 1178

CAA-TAA AATCGGAAGGAGGTTACGTTTGCTGATGAGTTGCCATAGCAGCTC

ACTAACCTTGGAAGAATCCAAGCGGCAAAAG

AGCAAACGTAACCTCCT 1179

AGGAGGTTACGTTTGCT 1180 2,4 DB resistance GCTGCTATGGCAACTCATCAGCAAACGCAACCTCCTTCCGATTTT | 1181

Pex14 CCCGCTCTTGCCGATTAAAATTCCCAGATTCCAGGTTCAATTTACA

Arabidopsis thaliana | CCTTCTAATCATTATTTCTTAATTITTCTT

Glu19Term AAGAAAAATTAAGAAATAATGATTAGAAGGTGTAAATTGAACCTGG | 1182

GAA-TAA AATCTGGGAATTTTAATCGGCAAGAGCGGGAAAATCGGAAGGAG

GTTGCGTTTGCTGATGAGTTGCCATAGCAGC

® TTGCCGATTAAAATTCC 1183

GGAATTTTAATCGGCAA 1184 2,4 DB resistance GCAACTCATCAGCAAACGCAACCTCCTTCCGATTTTCCCGCTCTT 1185

Pex14 GCCGATGAAAATTCCTAGATTCCAGGTTCAATTTACACCTTCTAAT

Arabidopsis thaliana | CATTATTTCTTAATTTTTCTTTGGTGGATT

GIn22Term AATCCACCAAAGAAAAATTAAGAAATAATGATTAGAAGGTGTAAAT | 1186

CAG-TAG TGAACCTGGAATCTAGGAATTTTCATCGGCAAGAGCGGGAAAATC

GGAAGGAGGTTGCGTTTGCTGATGAGTTGC

AAAATTCCTAGATTCCA 1187

TGGAATCTAGGAATTTT 1188

I Ft) WS 1412001 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Example 8

Production of albino mutants for the analysis of photosynthetic processes

Plant productivity is limited by resources available and the ability of plants to hamess these resources. The conversion of light to chemical energy, which is then used to synthesize carbohydrates, fatty acids, sugars, amino acids and other compounds, requires a complex system which combines the light harvesting apparatus of pigments and proteins. The value of light energy to the plant can only be realized when it is efficiently converted into chemical energy by photosynthesis and fed into various biochemical processes. Significant effort has therefore been directed at studying photosynthetic processes in plants in ® order to improve productivity and/or the efficiency of photosynthesis. The analysis of the photosynthetic process is substantially aided by the ability to produce albino plants.

The attached table discloses exemplary oligonucleotide base sequences which can be used to generate site-specific mutations in genes involved in starch metabolism. 1912/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Table 18

Oligonucleotides to produce albino plants

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

White leaves TTCTTTCCTGTGAAATTATCTGCTCAAATCTTTGGTTCCTGACGGAG| 1189

Immutans ATGGCGGCGATTTGAGGCATCTCCTCTGGTACGTTGACGATTTCA

Arabidopsis thaliana |CGGCCTTTGGTTACTCTTCGACGCTCTAG

Serb5Term CTAGAGCGTCGAAGAGTAACCAAAGGCCGTGAAATCGTCAACGTA | 1190

TCA-TGA CCAGAGGAGATGCCTCAAATCGCCGCCATCTCCGTCAGGAACCAA

AGATTTGAGCAGATAATTTCACAGGAAAGAA

GGCGATTTGAGGCATCT 1191 o AGATGCCTCAAATCGCC 1192

Immutans TCTCCTCTGGTACGTAGACGATTTCACGGCCTTTGGTTACTCTTCG

Arabidopsis thaliana |ACGCTCTAGAGCCGCCGTTTCGTACAGCTC

Leu12Term GAGCTGTACGAAACGGCGGCTCTAGAGCGTCGAAGAGTAACCAAA] 1194

TTG-TAG GGCCGTGAAATCGTCTACGTACCAGAGGAGATGCCTGAAATCGCC

GCCATCTCCGTCAGGAACCAAAGATTTGAGC

TGGTACGTAGACGATTT

AAATCGTCTACGTACCA 1196

White leaves : AGA A A i 0

Immutans GTACGTTGACGATTTGACGGCCTTTGGTTACTCTTCGACGCTCTAG

Arabidopsis thaliana |AGCCGCCGTTTCGTACAGCTCCTCTCACCG

Ser15Term CGGTGAGAGGAGCTGTACGAAACGGCGGCTCTAGAGCGTCGAAG | 1198

TCA-TGA AGTAACCAAAGGCCGTCAAATCGTCAACGTACCAGAGGAGATGCC [ TGAAATCGCCGCCATCTCCGTCAGGAACCAAA

GACGATTTGACGGCCTT 1199

AAGGCCGTCAAATCGTC 1200

Immutans CCTTTGGTTACTCTTIGACGCTCTAGAGCCGCCGTTTCGTACAGCT

Arabidopsis thaliana |CCTCTCACCGATTGCTTCATCATCTTCCTC

Arg22Term GAGGAAGATGATGAAGCAATCGGTGAGAGGAGCTGTACGAAACG 1202

CGA-TGA GCGGCTCTAGAGCGTCAAAGAGTAACCAAAGGCCGTGAAATCGTC

AACGTACCAGAGGAGATGCCTGAAATCGCCGC

, TTACTCTTIGACGCTCT 1203

AGAGCGTCAAAGAGTAA 1204 1112/01 01:47 pm 03137.003 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797 \W ite leaves A A A A K A A (} immutans CTCTTCGACGCTCTIGAGCCGCCGTTTCGTACAGCTCCTCTCACC

Arabidopsis thaliana |GATTGCTTCATCATCTTCCTCTCTCTTCTC

Arg25Term GAGAAGAGAGAGGAAGATGATGAAGCAATCGGTGAGAGGAGCTG | 1206

AGA-TGA TACGAAACGGCGGCTCAAGAGCGTCGAAGAGTAACCAAAGGCCG : TGAAATCGTCAACGTACCAGAGGAGATGCCTGA

GACGCTCTTGAGCCGCC 1207

GGCGGCTCAAGAGCGTC 1208

White leaves GATTCTTGTGGGAAGGAAGAAGGATCAAGAATGGCGATTTCGATT | 1209

Immutans TCTGCTATGAGTTTTIGAACCTCAGTTTCTTCATATTCTTGTTTTAG

Lycopersicon AGCTAGGAGTTTTGAGAAGTCATCAGTTT esculentum AAACTGATGACTTCTCAAAACTCCTAGCTCTAAAACAAGAATATGA | 1210

Gly11Term AGAAACTGAGGTTCAAAAACTCATAGCAGAAATCGAAATCGCCATT ) GGA-TGA CTTGATCCTTCTTCCTTCCCACAAGAATC

TGAGTTTTIGAACCTCA 1211

TGAGGTTCAAAAACTCA 1212

White leaves GTGGGAAGGAAGAAGGATCAAGAATGGCGATTTCGATTTCTGCTA | 1213 immutans TGAGTTTTGGAACCTGAGTTTCTTCATATTCTTGTTTTAGAGCTAGG

Lycopersicon AGTTTTGAGAAGTCATCAGTTTTATGCAA esculentum TTGCATAAAACTGATGACTTCTCAAAACTCCTAGCTCTAAAACAAG | 1214

Ser13Term AATATGAAGAAACTCAGGTTCCAAAACTCATAGCAGAAATCGAAAT

TCA-TGA CGCCATTCTTGATCCTTCTTCCTTCCCAC

TGGAACCTGAGTTICTT 1215

AAGAAACTCAGGTTCCA 1216

White leaves AAGAAGGATCAAGAATGGCGATTTCGATTTCTGCTATGAGTTITGG | 1217

Immutans AACCTCAGTTTCTTGATATTCTTGTTTTAGAGCTAGGAGTTTTGAGA

Lycopersicon AGTCATCAGTTTTATGCAATTCCCAGAA esculentum TTCTGGGAATTGCATAAAACTGATGACTTCTCAAAACTCCTAGCTC | 1218 ® Ser16Term TAAAACAAGAATATCAAGAAACTGAGGTTCCAAAACTCATAGCAGA

TCA-TGA AATCGAAATCGCCATTCTTGATCCTTCTT

AGTTTCTTGATATTCTT 1219

AAGAATATCAAGAAACT 1220

White leaves AGGATCAAGAATGGCGATTTCGATTTCTGCTATGAGTTTTGGAACC | 1221

Immutans TCAGTTTCTTCATAGTCTTGTTTTAGAGCTAGGAGTTTTGAGAAGTC

Lycopersicon ATCAGTTTTATGCAATTCCCAGAACCCA esculentum TGGGTTCTGGGAATTGCATAAAACTGATGACTTCTCAAAACTCCTA | 1222 : Tyri7Term GCTCTAAAACAAGACTATGAAGAAACTGAGGTTCCAAAACTCATAG

TAT-TAG CAGAAATCGAAATCGCCATTCTTGATCCT

TCTTCATAGTCTTGTTT 1223

AAACAAGACTATGAAGA 1224 11/12/01 01:47 pm 03137.008 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

White leaves AAGAATGGCGATTTCGATTTCTGCTATGAGTTTTGGAACCTCAGTT | 1225

Immutans TCTTCATATTCTTGATTTAGAGCTAGGAGTTTTGAGAAGTCATCAGT

Lycopersicon TTTATGCAATTCCCAGAACCCATGTCGG esculentum CCGACATGGGTTCTGGGAATTGCATAAAACTGATGACTTCTCAAAA | 1226

Cys19Term CTCCTAGCTCTAAATCAAGAATATGAAGAAACTGAGGTTCCAAAAC

TGT-TGA TCATAGCAGAAATCGAAATCGCCATTCTT

TATTCTTGATTTAGAGC 1227

GCTCTAAATCAAGAATA 1228

White leaves CGCGTCCGATAAAAAAATCAAGAATGGCGATTTCCATATCTGCTAT | 1229

Immutans GAGTTTTCGAACTTGAGTTTCTTCTTCATATTCAGCATTTTTGTGCA

Capsicum annuum ATTCCAAGAACCCATTTTGTTTGAATTC

Ser13Term GAATTCAAACAAAATGGGTTCTTGGAATTGCACAAAAATGCTGAAT | 1230

TCA-TGA ATGAAGAAGAAACTCAAGTTCGAAAACTCATAGCAGATATGGAAAT @ CGCCATTCTTGATTTTTTTATCGGACGCG

TCGAACTTGAGTTTCTT 1231

AAGAAACTCAAGTTCGA 1232

White leaves AAAAATCAAGAATGGCGATTTCCATATCTGCTATGAGTTTTCGAAC | 1233 immutans TTCAGTTTCTTCTTGATATTCAGCATTTTTGTGCAATTCCAAGAACC

Capsicum annuum CATTTTGTTTGAATTCTCTATTTTCACT 16 Ser17Term AGTGAAAATAGAGAATTCAAACAAAATGGGTTCTTGGAATTGCACA | 1234

TCA-TGA AAAATGCTGAATATCAAGAAGAAACTGAAGTTCGAAAACTCATAGC

AGATATGGAAATCGCCATTCTTGATTTTT

TTCTTCTTGATATTCAG

CTGAATATCAAGAAGAA 1236

White leaves CAAGAATGGCGATTTCCATATCTGCTATGAGTTTTCGAACTTCAGT | 1237

Immutans TTCTTCTTCATATTGAGCATTTTTGTGCAATTCCAAGAACCCATTTT

Capsicum annuum GTTTGAATTCTCTATTTTCACTTAGGAA @ Ser19Term TTCCTAAGTGAAAATAGAGAATTCAAACAAAATGGGTTCTTGGAAT | 1238

TCA-TGA TGCACAAAAATGCTCAATATGAAGAAGAAACTGAAGTTCGAAAACT + |CATAGCAGATATGGAAATCGCCATTCTTG

TTCATATTGAGCATTTT 1239

AAAATGCTCAATATGAA 1240

White leaves CGATTTCCATATCTGCTATGAGTTTTCGAACTTCAGTTTCTTCTTCA | 1241

Immutans TATTCAGCATTTTAGTGCAATTCCAAGAACCCATTTTGTTTGAATTC

Capsicum annuum TCTATTTTCACTTAGGAATTCTCATAG

Leu21Term CTATGAGAATTCCTAAGTGAAAATAGAGAATTCAAACAAAATGGGT | 1242

TTG-TAG TCTTGGAATTGCACTAAAATGCTGAATATGAAGAAGAAACTGAAGT

TCGAAAACTCATAGCAGATATGGAAATCG

AGCATTTTAGTGCAATT 1243

AATTGCACTAAAATGCT 1244 11112001 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

White leaves TTCCATATCTGCTATGAGTTTTCGAACTTCAGTTTCTTCTTCATATT | 1245 immutans CAGCATTTTTGTGAAATTCCAAGAACCCATTTTGTTTGAATTCTCTA

Capsicum annuum TTTTCACTTAGGAATTCTCATAGAACT

Cys22Term AGTTCTATGAGAATTCCTAAGTGAAAATAGAGAATTCAAACAAAAT | 1246

TGC-TGA GGGTTCTTGGAATTTICACAAAAATGCTGAATATGAAGAAGAAACTG

AAGTTCGAAAACTCATAGCAGATATGGAA

TTTTTGTGAAATTCCAA 1247

TTGGAATTTCACAAAAA 1248

White leaves TTCGGCACGAGGGAGAAGGAGCAGACCGAGGTGGCCGTCGAGG | 1249

Immutans AGTCCTTCCCCTTCAGGTAGACGGCTCCTCCTGACGAGCCACTGG

Oryza sativa TCACCGCCGAGGAGAGCTGGGTGGTTAAGCTCG

Glu22Term CGAGCTTAACCACCCAGCTCTCCTCGGCGGTGACCAGTGGCTCGT | 1250 é GAG-TAG CAGGAGGAGCCGTCTACCTGAAGGGGAAGGACTCCTCGACGGCC

ACCTCGGTCTGCTCCTTCTCCCTCGTGCCGAA

CCTTCAGGTAGACGGCT 1251

AGCCGTCTACCTGAAGG 1252

Immutans GACGGCTCCTCCTGACTAGCCACTGGTCACCGCCGAGGAGAGCT

Oryza sativa GGGTGGTTAAGCTCGAGCAGTCCGTGAACATTT

Glu28Term AAATGTTCACGGACTGCTCGAGCTTAACCACCCAGCTCTCCTCGG | 1254

CAG-TAG CGGTGACCAGTGGCTAGTCAGGAGGAGCCGTCTCCCTGAAGGGG

AAGGACTCCTCGACGGCCACCTCGGTCTGCTC

CTCCTGACTAGCCACTG 1255

CAGTGGCTAGTCAGGAG 1256

Immutans GCCACTGGTCACCGCCTAGGAGAGCTGGGTGGTTAAGCTCGAGC

Oryza sativa AGTCCGTGAACATTTTCCTCACGGAGTCAGTCA ® Glu34Term TGACTGACTCCGTGAGGAAAATGTTCACGGACTGCTCGAGCTTAA | 1258

GAG-TAG CCACCCAGCTCTCCTAGGCGGTGACCAGTGGCTCGTCAGGAGGA

GCCGTCTCCCTGAAGGGGAAGGACTCCTCGAC

TCACCGCCTAGGAGAGC 1259

GCTCTCCTAGGCGGTGA 1260

White leaves GAGGAGTCCTTCCCCTTCAGGGAGACGGCTCCTCCTGACGAGCC | 1261

Immutans ACTGGTCACCGCCGAGTAGAGCTGGGTGGTTAAGCTCGAGCAGT

Oryza sativa CCGTGAACATTTTCCTCACGGAGTCAGTCATCA

Glu35Term TGATGACTGACTCCGTGAGGAAAATGTTCACGGACTGCTCGAGCT | 1262

GAG-TAG TAACCACCCAGCTCTACTCGGCGGTGACCAGTGGCTCGTCAGGA

GGAGCCGTCTCCCTGAAGGGGAAGGACTCCTC

CCGCCGAGTAGAGCTGG 1263

CCAGCTCTACTCGGCGG 1264 11/1201 01:47 pm 03137.008 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

White leaves CTTCCCCTTCAGGGAGACGGCTCCTCCTGACGAGCCACTGGTCAC | 1265

Immutans CGCCGAGGAGAGCTGAGTGGTTAAGCTCGAGCAGTCCGTGAACA . Oryza sativa TTTTCCTCACGGAGTCAGTCATCACGATACTT

Trp37Term AAGTATCGTGATGACTGACTCCGTGAGGAAAATGTTCACGGACTG | 1266

TGG-TGA CTCGAGCTTAACCACTCAGCTCTCCTCGGCGGTGACCAGTGGCTC : GTCAGGAGGAGCCGTCTCCCTGAAGGGGAAG

GAGAGCTGAGTGGTTAA 1267

TTAACCACTCAGCTCTC 1268

White leaves [TCCGGAGGAGGAAGGGGGATTCGACGAGGAGCTCACCCTCGCCG | 1269

Immutans GCGAGGACGGCGACTGAGTCGTCAGATTCGAGCAGTCCTTCAAC

Triticum aestivum GTATTCCTCACGGATACTGTCATCTTTATACTC

Trp22Term GAGTATAAAGATGACAGTATCCGTGAGGAATACGTTGAAGGACTG | 1270 ® TGG-TGA CTCGAATCTGACGACTCAGTCGCCGTCCTCGCCGGCGAGGGTGA

GCTCCTCGTCGAATCCCCCTTCCTCCTCCGGA

GGCGACTGAGTCGTCAG 1271

CTGACGACTCAGTCGCC 1272

Immutans ACGGCGACTGGGTCGTCTIGATTCGAGCAGTCCTTCAACGTATTCC

Triticum aestivum TCACGGATACTGTCATCTTTATACTCGATATTC

Arg25Term GAATATCGAGTATAAAGATGACAGTATCCGTGAGGAATACGTTGAA | 1274

AGA-TGA GGACTGCTCGAATCAGACGACCCAGTCGCCGTCCTCGCCGGCGA

GGGTGAGCTCCTCGTCGAATCCCCCTTCCTC

GGGTCGTCTIGATTCGAG 1275

CTCGAATCAGACGACCC

Immutans ACTGGGTCGTCAGATTCTAGCAGTCCTTCAACGTATTCCTCACGGA

Triticum aestivum TACTGTCATCTTTATACTCGATATTCTGTATC

J Glu27Term GATACAGAATATCGAGTATAAAGATGACAGTATCCGTGAGGAATAC | 1278

GAG-TAG GTTGAAGGACTGCTAGAATCTGACGACCCAGTCGCCGTCCTCGCC

GGCGAGGGTGAGCTCCTCGTCGAATCCCCC

TCAGATTCTAGCAGTCC 1279

GGACTGCTAGAATCTGA 1280 immutans GGTCGTCAGATTCGAGTAGTCCTTCAACGTATTCCTCACGGATACT

Triticum aestivum GTCATCTTTATACTCGATATTCTGTATCGTG } GIn28Term CACGATACAGAATATCGAGTATAAAGATGACAGTATCCGTGAGGAA| 1282

CAG-TAG TACGTTGAAGGACTACTCGAATCTGACGACCCAGTCGCCGTCCTC

GCCGGCGAGGGTGAGCTCCTCGTCGAATCC

GATTCGAGTAGTCCTTC 1283

GAAGGACTACTCGAATC 1284 1112/01 01:47 pm 03137.009 — [NY}793559.1

AMENDED SHEET

28-12-2001 01939797

White leaves CGAGCAGTCCTTCAACGTATTCCTCACGGATACTGTCATCTTTATA 1285

Immutans CTCGATATTCTGTAGCGTGACCGCGACTACGCAAGGTTCTTCGTG

Triticum aestivum CTCGAGACCATCGCCAGGGTGCCCTATTTC

Tyrd6Term GAAATAGGGCACCCTGGCGATGGTCTCGAGCACGAAGAACCTTG 1286

TAT-TAG CGTAGTCGCGGTCACGCTACAGAATATCGAGTATAAAGATGACAG : TATCCGTGAGGAATACGTTGAAGGACTGCTCG

ATTCTGTAGCGTGACCG 1287

CGGTCACGCTACAGAAT 1288 11/12/01 01:47 pm 03137.009 — [NY]793550.1

AMENDED SHEET

28-12-2001 01939797

Example 9

Altering amino acid content of plants

Another aim of biotechnology is to generate plants, especially crop plants, with added value traits. An example of such a trait is improved nutritional quality in food crops. For example, lysine, tryptophan and threonine, which are essential amino acids in the diet of humans and many animals, are limiting nutrients in most cereal crops. Consequently, grain-based diets, such as those based on com, barley, wheat, rice, maize, millet, sorghum, and the like, must be supplemented with more expensive synthetic amino acids or amino-acid-containing oilseed protein meals. Increasing the lysine content of these ® grains or of any of the feed component crops would result in significant added value.

Naturally occurring mutants of plants that have different levels of particular essential amino acids have been identified. However, these mutants are generally not the result of increased free amino acid, but are instead the result of shifts in the overall protein profile of the grain. For example, in maize, reduced levels of lysine-deficient endosperm proteins (prolamines) are complemented by elevated levels of more lysine-rich proteins (albumins, globulins and glutelins). While nutritionally superior, these mutants are associated with reduced yields and poor grain quality, limiting their agronomic usefulness.

An alternative approach is to generate plants with mutations that render key amino acid biosynthetic enzymes insensitive to feedback inhibition. Many such mutations are known and mutation results in increased free amino acid. The increased production can optionally be coupled to increased expression of an abundant storage protein comprising the chosen amino acid. Alternatively, a normally [ abundant protein can be engineered to contain more of the target amino acid.

The attached table discloses exemplary oligonucleotide base sequences which can be used to generate site-specific mutations that remove feedback inhibition in plant amino acid biosynthetic enzymes. 03437008 NYITO3s5o.

AMENDED SHEET

28-12-2001 01939797

Table 19

Genome-Altering Oligos Conferring Amino Acid Overproduction

Phenotype, Gene, . Plant & Targeted Altering Oligos

Alteration ]

CGS GAGCATTAAAGCCCATAGAAACTGTAGCAACATCGGTGTTGCACA

Arabidopsis thaliana | GATCGTGGCGGCTAAGTGGTCCAACAACCC

Arg77His GGGTTGTTGGACCACTTAGCCGCCACGATCTGTGCAACACCGAT 1290

ATTAGGAGGAAATCTTAAGATCCTGGAGGATA

°

Met Overproduction | TCTTAAGATITCCTCCTAATTTCGTCCGTCAGCTGAGCATTAAAGC on

Arabidopsis thaliana | CTAAGTGGTCCAACAACCCATCCTCCGCGTT

Ser81Asn AACGCGGAGGATGGGTTGTTGGACCACTTAGCCGCCACGATCTG | 1294

TGACGGACGAAATTAGGAGGAAATCTTAAGA et Overproduction | TTTCCTCCTAATTICGTCCGTCAGCTGAGCATTAAA CGTAGAA 297

TT re

Arabidopsis thaliana | CCAACAACCCATCCTCCGCGTTACCTTCGG

Gly84Ser CCGAAGGTAACGCGGAGGATGGGTTGTTGGACCACTTAGCCGCC | 1298 } TGCTCAGCTGACGGACGAAATTAGGAGGAAA ®

Met Overproduction TCCTCCTAATTTCGTCCGTCAGCTGAGCATTAAAGCCCGTAGAAA | 1301

Te ee

Arabidopsis thaliana | CAACAACCCATCCTCCGCGTTACCTTCGGC

Gly84Asp GCCGAAGGTAACGCGGAGGATGGGTTGTTGGACCACTTAGCCGC | 1302

ATGCTCAGCTGACGGACGAAATTAGGAGGAA

1111201 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Es

Alteration

Vet Overproduction [TATCGTCACTCATCCTCCGCTTCCCT AACT ,CGCCA

EB

Fragraria vesca CAGATCGTCGCGGCTTCGTGGTCCAACAAAGA

CGC-CAC GTTGCTGCAGTTGCGGIGGGCCTTGGTGCTGAGCTGGCGGACGA

AGTTGGGAGGGAAGCGGAGGATGAGTGACGATA

CGS CCCGCCGCAACTGCAACAACATCGGCGTCGCGCAGATCGTCGCG

® Fragraria vesca GCTTCGTGGTCCAACAAAGACTCCGACCTTTC

AGC-AAC CGCGACGCCGATGTTGITGCAGTTGCGGCGGGCCTTGGTGCTGA

GCTGGCGGACGAAGTTGGGAGGGAAGCGGAGGA et Overproduction | TTCCCTCCCAACTTCGTCCGCCAGCTCAGCACCAAGGCCCGCCG | 1313 mem

Fragraria vesca GGTCCAACAAAGACTCCGACCTTTCGGCGGTGC

Gly80Ser GCACCGCCGAAAGGTCGGAGTCTTTGTTGGACCACGAAGCCGCG | 1314

GGTGCTGAGCTGGCGGACGAAGTTGGGAGGGAA

Met Overproduction CCCTCCCAACTTCGTCCGCCAGCTCAGCACCAA 7 « ETT Eom

Fragraria vesca GTCCAACAAAGACTCCGACCTTTCGGCGGTGCC

GGC-GAC GACGATCTGCGCGACGICGATGTTGCTGCAGTTGCGGCGGGCCT

TGGTGCTGAGCTGGCGGACGAAGTTGGGAGGGA

Met Overproduction | TCTCCTCCCTCATCCTCCGCTTCCCTCCCAACTTCCAGCGCCAGC 3

Te ee

Glycine max CAAATCGTCGCCGCTTCGTGGTCGAACAACAG

Arg68His CTGTTGTTCGACCACGAAGCGGCGACGATTTGCGCGACGCCGAT | 1322

AGTTGGGAGGGAAGCGGAGGATGAGGGAGGAGA

1142001 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration _ Jereooeerceceniete 1 faa : Viet Overproduction T T AACTTCCA A AAGCACCAAC 25

CGS CGCGCCGCAACTGCAACAACATCGGCGTCGCGCAAATCGTCGCC

Ser72Asn GGAGAGTTGTCGCTGTTGTTCGACCACGAAGCGGCGACGATTTG | 1326

GCTGGCGCTGGAAGTTGGGAGGGAAGCGGAGGA

CGS CAACTGCAGCAACATCAGCGTCGCGCAAATCGTCGCCGCTTCGT

Glycine max GGTCGAACAACAGCGACAACTCTCCGGCCGCCS

Gly75Ser CGGCGGCCGGAGAGTTGTCGCTGTTGTTCGACCACGAAGCGGCG | 1330

GGTGCTTAGCTGGCGCTGGAAGTTGGGAGGGAA

Met Overproduction SCTCCCAACTTCCAGCGCCAGCTAAGCACCAAGGCGCGC 1333

Ee

Glycine max GTCGAACAACAGCGACAACTCTCCGGCCGCCGG

GGC-GAC GACGATTTGCGCGACGICGATGTTGCTGCAGTTGCGGCGCGCCT

TGGTGCTTAGCTGGCGCTGGAAGTTGGGAGGGA

® Met Overproduction | TGTCTTCTCTGATTTTCAGGTTTCCTCCTAATTTCGTGAGGCAGCT 337

CGS AAGCATTAAGGCTCACAGGAATTGCAGCAATATTGGCGTGGCTCA TT

Solanum tuberosum | AGTTGTGGCGGCTTCCTGGTCTAACAACCA

Arg70His TGGTTGTTAGACCAGGAAGCCGCCACAACTTGAGCCACGCCAATA

AGG-CAC TTGCTGCAATTCCTGTGAGCCTTAATGCTTAGCTGCCTCACGAAAT

TAGGAGGAAACCTGAAAATCAGAGAAGACA

CGS TAGGAGGAATTGCAACAATATTGGCGTGGCTCAAGTTGTGGCGG CT

Solanum tuberosum

AGC-AAC AGCCACGCCAATATTGITGCAATTCCTCCTAGCCTTAATGCTTAGC

TGCCTCACGAAATTAGGAGGAAACCTGAAAA

1112001 01:47 pm . 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene, . Alteration — [GMATIGCAACMIATIG [fas [oummmeme ar et Overproduction | TTTCCTCCTAATTTCGTGAGGCAGCTAAGCATTAAGGCTAGGA 345

CGS AATTGCAGCAATATTAGCGTGGCTCAAGTTGTGGCGGCTTCCTGG oT

Gly77Ser GAGTGAATTCAGGACCGGCTTGGTTGTTAGACCAGGAAGCCGCC | 1346

GGC-AGC ACAACTTGAGCCACGCTAATATTGCTGCAATTCCTCCTAGCCTTAA 0

PY

Met Overproduction TTICCTCCTAATTTCGTGAGGCAGCTAAGCATTAAGGCTAGGAGGA 349

CGS ATTGCAGCAATATTGACGTGGCTCAAGTTGTGGCGGCTTCCTGET TT

Gly77Asp GGAGTGAATTCAGGACCGGCTTGGTTGTTAGACCAGGAAGCCGC

GGC-GAC CACAACTTGAGCCACGTCAATATTGCTGCAATTCCTCCTAGCCTTA et Overproduction TTCCTCTCTTATCCTTCGCTTTCCTCCCAACTITTGICCGTCAGCT | 1353

CGS CAGCACCAAGGCTCGCCACAACTGCAGCAACATTGGTGTCGCAC oT crystallinum GAGTTGTTGGACCAGGAGGCAGCGACGACCTGTGCGACACCAAT | 1354

Arg73His GTTGCTGCAGTTGTGGCGAGCCTTGGTGCTGAGCTGACGGACAA ES ° et Overproduction” | TCCTICGCTTTCCTCCCAACTTTGICCGTCAGCTCAGCACCAAGG | 1357

CGS CTCGCCGCAACTGCAACAACATTGGTGTCGCACAGGTCGTCGCT TT crystallinum GCGCCGGCATCGGAGTTGTTGGACCAGGAGGCAGCGACGACCT | 1358

Ser77Asn GTGCGACACCAATGTTGITGCAGTTGCGGCGAGCCTTGGTGCTG BB 1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene, ow BP : Alteration

Met Overproduction [| TTTCCTCCCAACTTTGTCCGTCAGCTCAGCACCAAGGCTC C

EEE

Mesembryanthemum | GTCCAACAACTCCGATGCCGGCGCCACCTCTT

Gly80Ser GACGACCTGTGCGACACTAATGTTGCTGCAGTTGCGGCGAGCCT

GGT-AGT TGGTGCTGAGCTGACGGACAAAGTTGGGAGGAAA ® Mesembryanthemum | CCAACAACTCCGATGCCGGCGCCACCTCTTG crystallinum CAAGAGGTGGCGCCGGCATCGGAGTTGTTGGACCAGGAGGCAG

Gly80Asp CGACGACCTGTGCGACATCAATGTTGCTGCAGTTGCGGCGAGCC et Overproduction | CCTCTGCTACCATCCTCCGCTTTCCGCCAAACTTTGTCCGCCAGC | 1369

CGS TTAGCACCAAGGCACACCGCAACTGCAGCAACATCGGCGTCGCG

Arg41His GGGCAGTCGGACCACGCGGCGGCGACGATCTGCGCGACGCCGA | 1370

AAGTTTGGCGGAAAGCGGAGGATGGTAGCAGAGG

Met Overproduction | TCCTCCGCTTTCCGCCAAACTTTGTCCGCCAGCTTAGCACCAAGG | 1373 o [FT Emm

Zea mays GCCGCGTGGTCCGACTGCCCCGCCGCTCGCCC

AGC-AAC GCGCGACGCCGATGTTGITGCAGTTGCGGCGTGCCTTGGTGCTA

AGCTGGCGGACAAAGTTTGGCGGAAAGCGGAGGA

CGS AACTGCAGCAACATCAGCGTCGCGCAGATCGTCGCCGCCGCGTG oT

Gly48Ser CTAAGTGGGGGCGAGCGGCGGGGCAGTCGGACCACGCGGCGG | 1378

GGC-AGC CGACGATCTGCGCGACGCIGATGTTGCTGCAGTTGCGGCGTGCC 1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

CC] CGCGACGCIGATGTTGC 1380 et Overproduction | TTCCGCCAAACTTTGTCCGCCAGCTTAGCACCAAGGCACGCCGCA | 138

CGS ACTGCAGCAACATCGACGTCGCGCAGATCGTCGCCGCCGCGTGG

Zea mays TCCGACTGCCCCGCCGCTCGCCCCCACTTAGG

Gly48Asp CCTAAGTGGGGGCGAGCGGCGGGGCAGTCGGACCACGCGGCG | 1382

GGC-GAC GCGACGATCTGCGCGACGTCGATGTTGCTGCAGTTGCGGCGTGC

CTTGGTGCTAAGCTGGCGGACAAAGTTTGGCGGAA

CAACATCGACGTCGCGC 1383

GCGCGACGICGATGTTG 1384

PY Met Overproduction | GTATGAATGATCTGTGGGTGAAACACTGTGGGATTAGTCATACA 85

TS GAAGTTTCAAGGATCGTGGAATGACTGTTTTGGTTAGTCAAGTTAA

Arabidopsis thaliana | TCGTCTGAGAAAGATGAAACGACCTGTGGT

Leu205Arg ACCACAGGTCGTTTCATCTTTCTCAGACGATTAACTTGACTAACCA | 1386

CTT-CGT AAACAGTCATTCCACGATCCTTGAAACTTCCTGTATGACTAATCCC

ACAGTGTTTCACCCACAGATCATTCATAC

CAAGGATCGTGGAATGA 1387

TCATTCCACGATCCTTG 1388

Met Overproduction GCATGACTGATTTGTGG AAACACTGTGGGATTA ATACTG 8

TS GTAGTTTTAAGGATCGTGGGATGACTGTTTTGGTGAGTCAAGTTAA

Solanum tuberosum | TCGCTTGCGGAAAATGCATAAACCGGTTGT

Leu198Arg ACAACCGGTTTATGCATTTTCCGCAAGCGATTAACTTGACTCACCA | 1390

CTT-CGT AAACAGTCATCCCACGATCCTTAAAACTACCAGTATGGCTAATCCC

ACAGTGTTTGACCCACAAATCAGTCATGC

TAAGGATCGTGGGATGA 1391 ® TCATCCCACGATCCTTA 1392

DHPS TAGGCAACACAGGAAACAACTCAACCAGAGAAGCCGTCCACGCA

Zea mays ACAGAACAGGGATTTGCTGTTGGCATGCATGC

Ser157Asn GCATGCATGCCAACAGCAAATCCCTGTTCTGTTGCGTGGACGGCT | 1394

AGC-AAC TCTCTGGTTGAGTTGITTCCTGTGTTGCCTATCACTTTGATTCTAG

AGCCAAAGCAGTTCACTGTGTGCCCAATGA

CACAGGAAACAACTCAA 1395

TTGAGTTGITTCCTGTG 1396

Lys Overproduction TCTAGAATCAAAGTGATA AACACAGGAAGCAACTCAACCA | 1397

DHPS GAGAAGCCGTCCACGAAACAGAACAGGGATTTGCTGTTGGCATG

Zea mays CATGCGGCTCTCCACATCAATCCTTACTACGG } Ala166Val CCGTAGTAAGGATTGATGTGGAGAGCCGCATGCATGCCAACAGC | 1398

GCA-GAA AAATCCCTGTTCTGTTTICGTGGACGGCTTCTCTGGTTGAGTTGCTT

CCTGTGTTGCCTATCACTTTGATTCTAGAGC

1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

CGTCCACGAAACAGAAC 1399 : GTTCTGTTICGTGGACG 1400 ys Overproduction GGCTCTAGAATCAAAGTGATAGGCAACACAGGAAGCAACTCAACC | 140

DHPS AGAGAAGCCGTCCACACAACAGAACAGGGATTTGCTGTTGGCAT

Zea mays GCATGCGGCTCTCCACATCAATCCTTACTACG :

Ala166Thr CGTAGTAAGGATTGATGTGGAGAGCCGCATGCATGCCAACAGCA | 1402

GCA-ACA AATCCCTGTTCTGTTGIGTGGACGGCTTCTCTGGTTGAGTTGCTTC

CTGTGTTGCCTATCACTTTGATTCTAGAGCC

CCGTCCACACAACAGAA 1403 ® TTCTGTTGIGTGGACGG 1404

Lys Overproduction ATTGGGCATACAGTTAACTGCTTTGGCACTAAAATTAAAGT 405

DHPS CGGCAACACAGGAAATAACTCAACAAGGGAGGCTATTCACGCAAC

Oryza sativa TGAGCAGGGATTCGCTGTAGGTATGCACGC

Ser124Asn GCGTGCATACCTACAGCGAATCCCTGCTCAGTTGCGTGAATAGCC | 1406

AGT-AAT TCCCTTGTTGAGTTATTTCCTGTGTTGCCGACCACTTTAATTTITAGT

GCCAAAGCAGTTAACTGTATGCCCAATAA

CACAGGAAATAACTCAA 1407

TTGAGTTATTTCCTGTG 1408

TI

DHPS GGGAGGCTATTCACGTAACTGAGCAGGGATTCGCTGTAGGTATG

Oryza sativa CACGCGGCTCTCCACATCAATCCTTACTACGG

Ala133val CCGTAGTAAGGATTGATGTGGAGAGCCGCGTGCATACCTACAGC | 1410 16 GCA-GTA GAATCCCTGCTCAGTTACGTGAATAGCCTCCCTTGTTGAGTTACTT

CCTGTGTTGCCGACCACTTTAATTTTAGTGC

® TATTCACGTAACTGAGC 1411

GCTCAGTTACGTGAATA 1412

DHPS AGGGAGGCTATTCACACAACTGAGCAGGGATTCGCTGTAGGTAT

Oryza sativa GCACGCGGCTCTCCACATCAATCCTTACTACG

Ala133Thr CGTAGTAAGGATTGATGTGGAGAGCCGCGTGCATACCTACAGCG | 1414

GCA-ACA AATCCCTGCTCAGTTGIGTGAATAGCCTCCCTTGTTGAGTTACTTC

CTGTGTTGCCGACCACTTTAATTTTAGTGCC

CTATTCACACAACTGAG 1415 : CTCAGTTGIGTGAATAG 1416

DHPS 1 AGGCAACACGGGAAATAACTCAACCAGAGAAGCTGTTCACGCGA iticum aestivum CAGAGCAGGGATTTGCTGTTGGCATGCATGC

Ser165Asn

AGT-AAT 11/4201 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene, )

Plant & Targeted Altering Oligos

Alteration

GCATGCATGCCAACAGCAAATCCCTGCTCTGTCGCGTGAACAGCT | 1418

TCTCTGGTTGAGTTATTTCCCGTGTTGCCTATCACTTTAATGTTGG

CTCCAAAGCAGTTAACAGTATGCCCGATGA

CACGGGAAATAACTCAA 1419

TTGAGTTAITTCCCGTG 1420 ys Overproduction AGCCAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAACCA | 1421

DHPS 1 GAGAAGCTGTTCACGTGACAGAGCAGGGATTTGCTGTTGGCATG

Triticum aestivum CATGCAGCTCTTCATGTCAATCCTTACTACGG

Ala174val CCGTAGTAAGGATTGACATGAAGAGCTGCATGCATGCCAACAGCA | 1422

GCG-GTG AATCCCTGCTCTGTCACGTGAACAGCTTCTCTGGTTGAGTTACTTC ® CCGTGTTGCCTATCACTTTAATGTTGGCTC

TGTTCACGTIGACAGAGC 1423

GCTCTGTCACGTGAACA 1424

DHPS 1 AGAGAAGCTGTTCACACGACAGAGCAGGGATTTGCTGTTGGCAT

Triticum aestivum GCATGCAGCTCTTCATGTCAATCCTTACTACG

Aia174Thr CGTAGTAAGGATTGACATGAAGAGCTGCATGCATGCCAACAGCAA | 1426

GCG-ACG ATCCCTGCTCTGTCGTIGTGAACAGCTTCTCTGGTTGAGTTACTTCC

CGTGTTGCCTATCACTTTAATGTTGGCTCC

CTGTTCACACGACAGAG 1427

CTCTGTCGIGTGAACAG 1428

DHPS 2 AGGCAACACGGGAAATAACTCAACTAGAGAAGCGATTCACGCTTC

Triticum aestivum AGAGCAGGGATTTGCTGTTGGCATGCATGC

Ser154Asn GCATGCATGCCAACAGCAAATCCCTGCTCTGAAGCGTGAATCGCT | 1430 @ AGT-AAT TCTCTAGTTGAGTTATTTCCCGTGTTGCCTATCACTTTAATGTTAGT

TCCAAAGCAGTTAACAGTGTGCCCGATGA

CACGGGAAATAACTCAA 1431

TTGAGTTAITTCCCGTG 1432 ys Overproduction | GAACTAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAACTA | 1433

DHPS 2 GAGAAGCGATTCACGTITTCAGAGCAGGGATTTGCTGTTGGCATGC

Triticum aestivum ATGCAGCTCTCCATGTCAATCCTTACTATGG

Ala163Val CCATAGTAAGGATTGACATGGAGAGCTGCATGCATGCCAACAGCA | 1434

GCT-GTT AATCCCTGCTCTGAAACGTGAATCGCTTCTCTAGTTGAGTTACTTC

CCGTGTTGCCTATCACTTTAATGTTAGTTC

GATTCACGITTCAGAGC 1435

GCTCTGAAACGTGAATC 1436

M201 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Lys Overproduction GGAACTAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAAC 43 : DHPS 2 AGAGAAGCGATTCACACTTCAGAGCAGGGATTTGCTGTTGGCATG

Triticum aestivum CATGCAGCTCTCCATGTCAATCCTTACTATG

Ala163Thr CATAGTAAGGATTGACATGGAGAGCTGCATGCATGCCAACAGCAA | 1438

GCT-ACT ATCCCTGCTCTGAAGIGTGAATCGCTTCTCTAGTTGAGTTACTTCC

CGTGTTGCCTATCACTTTAATGTTAGTICC

CGATTCACACTTCAGAG 1439

CTCTGAAGIGTGAATCG 1440 ys Overproduction ATTGGGCATACTGTGAACTGCTTTGGCTCTAGAATTAAAGTGA iE

DHPS TAGGCAACACAGGAAATAACTCAACCAGAGAAGCTGTTCACGCAA

® Coix lacryma-jobi CAGAGCAGGGATTTGCTGTTGGCATGCATG

Ser154Asn CATGCATGCCAACAGCAAATCCCTGCTCTGTTGCGTGAACAGCTT | 1442

AGT-AAT CTCTGGTTGAGTTATTTCCTGTGTTGCCTATCACTTTAATTCTAGA

GCCAAAGCAGTTCACAGTATGCCCAATGAG

CACAGGAAATAACTCAA 1443

TTGAGTTATTTCCTGTG

Lys Overproduction CTCTAGAATTAAAGTGATAGGCAACACAGGAAGTAACTCAACCA | 1445

DHPS GAGAAGCTGTTCACGTAACAGAGCAGGGATTTGCTGTTGGCATGC

Coix lacryma-jobi ATGCAGCTCTCCACATCAATCCTTACTATGG

Ala163Val CCATAGTAAGGATTGATGTGGAGAGCTGCATGCATGCCAACAGCA | 1446

GCA-GTA AATCCCTGCTCTGTTACGTGAACAGCTTCTCTGGTTGAGTTACTTC

CTGTGTTGCCTATCACTTTAATTCTAGAGC

TGTTCACGTAACAGAGC

GCTCTGTTACGTGAACA 1448 ® ys Overproduction CTCTAGAATTAAAGTGATAGGCAACACAGGAAGTAACTCAA 1449

DHPS AGAGAAGCTGTTCACACAACAGAGCAGGGATTTGCTGTTGGCATG

Coix lacryma-jobi CATGCAGCTCTCCACATCAATCCTTACTATG

Ala163Thr CATAGTAAGGATTGATGTGGAGAGCTGCATGCATGCCAACAGCAA | 1450

GCA-ACA ATCCCTGCTCTGTTGTGTGAACAGCTTCTCTGGTTGAGTTACTTCC

TGTGTTGCCTATCACTTTAATTCTAGAGCC

CTGTTCACACAACAGAG 1451

CTCTGTTGIGTGAACAG 1452 ys Overproduction | TCATTGGTCACACAGTCAATTGI TTTGGAGGGTCCATCAAAGTCAT | 1453 . DHPS CGGGAACACTGGAAACAACTCCACAAGGGAAGCAATCCATGCAA

Nicotiana tabacum CTGAACAGGGATTTGCTGTAGGTATGCATGC

Ser136Asn GCATGCATACCTACAGCAAATCCCTGTTCAGTTGCATGGATTGCTT | 1454 "25 AGC-AAC CCCTTGTGGAGTTGITTCCAGTGTTCCCGATGACTTTGATGGACC

CTCCAAAACAATTGACTGTGTGACCAATGA

CACTGGAAACAACTCCA 1455 1412/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration [| TGGAGTTGITTCCAGTG 1456 : ys Overproduction | GAGGGTCCATCAAAGTCATCGGGAACACTGGAAGCAACTCCACAA] 145

DHPS GGGAAGCAATCCATGTAACTGAACAGGGATTTGCTGTAGGTATGC

Nicotiana tabacum ATGCAGCTCTTCACATTAATCCCTACTATGG

Ala145Val CCATAGTAGGGATTAATGTGAAGAGCTGCATGCATACCTACAGCA | 1458

GCA-GTA AATCCCTGTTCAGTTACATGGATTGCTTCCCTTGTGGAGTTGCTTC

CAGTGTTCCCGATGACTTTGATGGACCCTC

AATCCATGTAACTGAAC 1459

GTTCAGTTACATGGATT

® ys Overproduction 3GA TCCATCAAAGTCATCGGGAACACTGGAAGCAACTCCAC | 1461

DHPS AAGGGAAGCAATCCATACAACTGAACAGGGATTTGCTGTAGGTAT

Nicotiana tabacum GCATGCAGCTCTTCACATTAATCCCTACTATG

Ala145Thr CATAGTAGGGATTAATGTGAAGAGCTGCATGCATACCTACAGCAA | 1462

GCA-ACA ATCCCTGTTCAGTTGTATGGATTGCTTCCCTTGTGGAGTTGCTTCC

AGTGTTCCCGATGACTTTGATGGACCCTCC

CAATCCATACAACTGAA

TTCAGTTGTATGGATTG 1464

Lys Overproduction | TTATAGGCCATACCGTTAACTGTTTTGGCGGAAGCATCAAAGTCAT | 1465

DHPS TGGAAACACTGGAAACAATTCGACTAGAGAAGCAATCCACGCGAC

Arabidopsis thaliana | TGAACAAGGATTCGCGGTTGGAATGCATGC

Ser142Asn GCATGCATTCCAACCGCGAATCCTTGTTCAGTCGCGTGGATTGCT | 1466

AGC-AAC TCTCTAGTCGAATTGTTTCCAGTGTTTCCAATGACTTTGATGCTTC

CGCCAAAACAGTTAACGGTATGGCCTATAA

CACTGGAAACAATTCGA 1467 ® TCGAATTGTTTCCAGTG 1468

DHPS GAGAAGCAATCCACGTGACTGAACAAGGATTCGCGGTTGGAATG

Co Arabidopsis thaliana | CATGCTGCTCTTCATATAAACCCTTACTATGG

Ala151Val CCATAGTAAGGGTTTATATGAAGAGCAGCATGCATTCCAACCGCG | 1470

GCG-GTG AATCCTTGTTCAGTCACGTGGATTGCTTCTCTAGTCGAATTGCTTC

CAGTGTTTCCAATGACTTTGATGCTTCCGC

AATCCACGTGACTGAAC 1471

GTTCAGTCACGTGGATT 1472 :

DHPS AGAGAAGCAATCCACACGACTGAACAAGGATTCGCGGTTGGAAT

Arabidopsis thaliana | GCATGCTGCTCTTCATATAAACCCTTACTATG

Ala151Thr CATAGTAAGGGTTTATATGAAGAGCAGCATGCATTCCAACCGCGA | 1474

GCG-ACG ATCCTTGTTCAGTCGIGTGGATTGCTTCTCTAGTCGAATTGCTTCC

AGTGTTTCCAATGACTTTGATGCTTCCGCC

1411201 01:47 pm : 03137009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ‘ Alteration

CAATCCACACGACTGAA 1475

TTCAGTCGIGTGGATTG 1476 ys Overproduction | TTATT ATACAGTCAACTGTTTTGGTGGGAAAATTAAGGTTAT | 1477

DHPS TGGAAATACTGGAAACAACTCCACCAGGGAAGCAATTCATGCCAC

Glycine max TGAGCAGGGTTTTGCTGTTGGAATGCATGC

Ser103Asn GCATGCATTCCAACAGCAAAACCCTGCTCAGTGGCATGAATTGCT | 1478

AGC-AAC TCCCTGGTGGAGTTGITTCCAGTATTTCCAATAACCTTAATTTICC

CACCAAAACAGTTGACTGTATGAGCAATAA

TACTGGAAACAACTCCA 1479 ® TGGAGTTGITTCCAGTA 1480 ys Overproduction |G AAAATTAAGGTTATTGGAAATA AAGCAACTCCACCA 481

DHPS GGGAAGCAATTCATGICACTGAGCAGGGTTTTGCTGTTGGAATGC

Glycine max ATGCTGCCCTTCACATAAACCCTTACTATGG

Ala112Vval CCATAGTAAGGGTTTATGTGAAGGGCAGCATGCATTCCAACAGCA | 1482

GCC-GTC AAACCCTGCTCAGTGACATGAATTGCTTCCCTGGTGGAGTTGCTT

CCAGTATTTCCAATAACCTTAATTTTCCCAC

AATTCATGICACTGAGC 1483

GCTCAGTGACATGAATT 1484

Lys Overproduction | GGTGGGAAAATTAAGGTTATTGGAAATACTGGAAGCAACTCCACC | 1485

DHPS AGGGAAGCAATTCATACCACTGAGCAGGGTTTTGCTGTTGGAATG

Glycine max CATGCTGCCCTTCACATAAACCCTTACTATG

Ala112Thr CATAGTAAGGGTTTATGTGAAGGGCAGCATGCATTCCAACAGCAA | 1486

GCC-ACC AACCCTGCTCAGTGGTATGAATTGCTTCCCTGGTGGAGTTGCTTC

CAGTATTTCCAATAACCTTAATTTTCCCACC

® CAATTCATACCACTGAG 1487

CTCAGTGGTIATGAATTG 1488 rp Overproduction | CTTGCAGGAGACATATTTCAGATC TGAGTCAACGTTITGA 48

AS CGGCGAACATTTGCAAACCCCTTTGAAGTTTATAGAGCACTAAGA

Arabidopsis thaliana | GTTGTGAATCCAAGTCCGTATATGGGTTATT

Asp341Asn AATAACCCATATACGGACTTGGATTCACAACTCTTAGTGCTCTATA | 1490

GAC-AAC AACTTCAAAGGGGTITGCAAATGTTCGCCGCTCAAAACGTTGACT

CAGCACGATCTGAAATATGTCTCCTGCAAG

CATTTGCAAACCCCTTT 1491

AAAGGGGTITGCAAATG 1492 rp Overproduction | GCTGCAGGAGACATATTTCAAATCGTTTTAAGTCAACGCTTTGAGA | 1493 : AS GAAGAACATTTGCTAACCCATTTGAAGTGTACAGAGCATTAAGAAT

Nicotiana tabacum TGTGAATCCAAGCCCATATATGACTTACA

Asp326Asn

GAC-AAC 1112001 01:47 pm 03137.009 —~ [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

TGTAAGTCATATATGGGCTTGGATTCACAATTCTTAATGCTCTGTA | 1494

CACTTCAAATGGGTTAGCAAATGTTCTTCTCTCAAAGCGTTGACTT

AAAACGATTTGAAATATGTCTCCTGCAGC

CATTTGCTAACCCATTT 1495

AAATGGGTTAGCAAATG 1496

Trp Overproduction TAGCTGGTGACATTTTTCAAGTAGTCTTAAGCCAGCGTTTTGAGA | 1497

AS GGCGTACATTTGCTAACCCCTTTGAGGTGTACCGTGCATTGCGTA

Oryza sativa TTGTCAATCCTAGTCCTTATATGGCCTATC

Asp323Asn GATAGGCCATATAAGGACTAGGATTGACAATACGCAATGCACGGT | 1488

GAC-AAC ACACCTCAAAGGGGTTAGCAAATGTACGCCTCTCAAAACGCTGGC ® TTAAGACTACTTGAAAAATGTCACCAGCTAG

CATTTGCTAACCCCTTT 1499

AAAGGGGTTAGCAAATG 1500

Trp Overproduction CTTGCTGGTGACATATTCCAGATCGTACTAAGTCAGCGTTTTGAAA | 1501

AS GGCGAACGTTCGCAAACCCATTTGAAATCTATAGATCACTGAGGA

Ruta graveolens TTGTTAATCCAAGCCCATATATGACTTATT

Asp354Asn AATAAGTCATATATGGGCTTGGATTAACAATCCTCAGTGATCTATA | 1502

GAC-AAC GATTTCAAATGGGTITGCGAACGTTCGCCTTTCAAAACGCTGACTT

AGTACGATCTGGAATATGTCACCAGCAAG

CGTTCGCAAACCCATTT 1503 ___ |AMATGGGTITGCGAACG | 1504

Trp Overproduction CTGGCTGGGGACATATTCCAGCTTGTCCTAAGTCAGCGTTTTGAA | 1505

AS CGGCGAACATTTGCAAATCCATTTGAAGTCTACCGAGCATTGAGA

Catharanthus roseus | ATTGTCAACCCAAGTCCATATATGACTTATT

Asp354Asn AATAAGTCATATATGGACTTGGGTTGACAATTCTCAATGCTCGGTA | 1506 @® GAT-AAT GACTTCAAATGGATTTGCAAATGTTCGCCGTTCAAAACGCTGACTT

AGGACAAGCTGGAATATGTCCCCAGCCAG

CATTTGCAAATCCATTT 1607

AAATGGATTTGCAAATG 1508 1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Example 10

Production of modified starch in pants

A principal aim of biotechnalogy is the improvement of crop plants for food value, agriculture, and to produce a range of plant-derived raw materials. Along with oils, fats and proteins, polysaccharides constitute the main raw materials derived from plants, and apart from cellulose, the storage polymer starch is the most important polysaccharide raw material. Starch is derived from a range of plants, but maize is the most important cultivated plant for the production of starch.

The polysaccharide starch is a polymer made up of glucose molecules. However, starch is not a homogeneous raw material and is, in fact, a highly complex mixture of various types of molecules ® which differ from each other, for example, in their degree of polymerization and in the degree of branching of the glucose chains. For example, amylose-starch is a basically non-branched polymer made up of a-1,4-glycosidically branched glucose molecules, and amylopectin-starch is a complex mixture of variously branched glucose chains. The branching results from additional -1,6-glycosidic linkages. In plants from which starch is typically isolated, for example maize or potato, the starch is approximately 25% amylose-starch and 75% amylopectin-starch.

In maize, various mutants in starch metabolism are known, for example waxy, sugary, shrunken and opaque-2. In addition to producing a modified starch, these mutations greatly improve grain quality in maize, and thus expand the use of maize not only as the food but also for the important industrial materials in food chemistry. It would therefore be advantageous to be able readily to obtain mutants in ( these genes in particular maize genotypes as well as other plants. Such plants can be obtained, for example, using traditional breeding methods and through specific genetic modification by means of recombinant DNA techniques.

The attached tables disclose exemplary oligonucleotide base sequences which can be used to generate site-specific mutations in genes involved in starch metabolism. 137.008 NY SsuD

AMENDED SHEET

28-12-2001 01939797

Table 20

Genome-Altering Oligos Conferring Increased Starch

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration _ GE——

Increased Starch GAACTTGAGACTGAGAAAAGGGATCCAAGGACAGTTGCTTCCATT | 1509

ADPGPP ATTCTTGGAGGTGGAAAAGGAACTCGACTCTTTCCTCTCACAAAA

Arabidopsis thaliana | CGCCGCGCCAAGCCTGCCGTTCCTATCGGGG

Ala99Lys CCCCGATAGGAACGGCAGGCTTGGCGCGGCGTTTTGTGAGAGGA | 1510

GCA-AAA AAGAGTCGAGTTCCTITTCCACCTCCAAGAATAATGGAAGCAACT

GTCCTTGGATCCCTTTTCTCAGTCTCAAGTTC

GAGGTGGAAAAGGAACT 1511 ® AGTTCCTITTCCACCTC increased Starc AAAACGCCGCGCCAAGCCTGCCGTTCCTATCGGGGGAGCCTA 1513

ADPGPP AGGTTGATAGATGTACTAATGAGCAATTGTATTAACAGCGGAATCA

Arabidopsis thaliana | ACAAAGTCTACATACTCACACAATATAACTC

Pro127Leu GAGTTATATTGTGTGAGTATGTAGACTTTGTTGATTCCGCTGTTAA | 1514

CCA-CTA TACAATTGCTCATTAGTACATCTATCAACCTATAGGCTCCCCCGAT

AGGAACGGCAGGCTTGGCGCGGCGTTTTG

AGATGTACTAATGAGCA

TGCTCATTAGTACATCT 1516

Increased Starch TCACACAATATAACTCAGCATCATTGAACAGGCATTTAGCCCGTGC | 1517

ADPGPP TTACAACTCCAATAATCTTGGCTTTGGAGATGGCTATGTTGAGGTT

: Arabidopsis thaliana | CTTGCGGCCACTCAAACGCCAGGAGAATC

Gly162Asn GATTCTCCTGGCGTTTGAGTGGCCGCAAGAACCTCAACATAGCCA | 1518

GGA-AAT TCTCCAAAGCCAAGATTATTGGAGTTGTAAGCACGGGCTAAATGC

CTGTTCAATGATGCTGAGTTATATTGTGTGA

® CTCCAATAATCTTGGCT 1519

AGCCAAGATTATTGGAG 1520

ADPGPP TTACAACTCCAATAACCTTGGCTTTGGAGATGGCTATGTTGAGGTT

Arabidopsis thaliana | CTTGCGGCCACTCAAACGCCAGGAGAATC

Gly162Asn GATTCTCCTGGCGTTTGAGTGGCCGCAAGAACCTCAACATAGCCA | 1522

GGA-AAC TCTCCAAAGCCAAGGTTATTGGAGTTGTAAGCACGGGCTAAATGC

CTGTTCAATGATGCTGAGTTATATTGTGTGA

: CTCCAATAACCTTGGCT 1523

AGCCAAGGTTATTGGAG 1524 1412/01 01:47 pm 03137.009 — [NY}793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos Sea) b __ Aeration |} 1

Increased Starch GTTTGAGAGAAGAAAGGTAGACCCGCAAAATGTGGCTGCAATCAT | 15256 | -

ADPGPP TCTAGGAGGAGGCAAAGGAGCTAAACTCTTCCCTCTTACAATGAG

Arabidopsis thaliana | AGCCGCAACACCAGCTGTAAATATTCATCTT

Asn100Lys AAGATGAATATTTACAGCTGGTGTTGCGGCTCTCATTGTAAGAGG | 1526

AAT-AAA GAAGAGTTTAGCTCCTTTGCCTCCTCCTAGAATGATTGCAGCCAC

ATTTTGCGGGTCTACCTTTCTTCTCTCAAAC

GGAGGCAAAGGAGCTAA 1527

TTAGCTCCITTGCCTCC 1528 [Increased Starch CTTGIGICTTCAAATTATGTTAGGTTCCT TGGATGCTACAG | 1528

ADPGPP GCTGATCGATATCCTGATGAGTAACTGTATTAACAGCTGCATCAAC

® Arabidopsis thaliana | AAGATATTTGTGCTGACACAGTTCAACTC

Pro128Leu GAGTTGAACTGTGTCAGCACAAATATCTTGTTGATGCAGCTGTTAA | 1530

CCG-CTG TACAGTTACTCATCAGGATATCGATCAGCCTGTAGCATCCACCAA

CAGGAACCTAACATAATTTGAAGACACAAG

CGATATCCIGATGAGTA 1531

TACTCATCAGGATATCG

Increased Starch TGACACAGTTCAACTCAGCTTCCCTTAATCGACATTTAGCACGAAC | 1533

ADPGPP TTATTTTGGGAATAATATAAACTTTGGAGGTGGTTTCGTAGAGGTA

Arabidopsis thaliana | CAAACACTATGACAATAATAACTCTCAGC

Gly163Asn GCTGAGAGTTATTATTGTCATAGTGTTTGTACCTCTACGAAACCAC | 1534

GGC-AAT CTCCAAAGTTTATATTATTCCCAAAATAAGTTCGTGCTAAATGTCG

ATTAAGGGAAGCTGAGTTGAACTGTGTCA

TGGGAATAATATAAACT

AGTTTATATTATTCCCA 1536 ®

ADPGPP TTATTTTGGGAATAACATAAACTTTGGAGGTGGTTTCGTAGAGGTA

Arabidopsis thaliana | CAAACACTATGACAATAATAACTCTCAGC

Gly163Asn GCTGAGAGTTATTATTGTCATAGTGTTTGTACCTCTACGAAACCAC | 1538

GGC-AAC CTCCAAAGTTTATGTTATTCCCAAAATAAGTTCGTGCTAAATGTCG

ATTAAGGGAAGCTGAGTTGAACTGTGTCA

TGGGAATAACATAAACT 1539

AGTTTATGITATTCCCA 1540 : ADPGPP TTCTAGGTGGTGGTAAAGGAACTCGTCTTTTTCCTCTTACAAGCA

Lycopersicon GAAGAGCTAAACCAGCTGTTCCTATTGGTGG esculentum CCACCAATAGGAACAGCTGGTTTAGCTCTTCTGCTTGTAAGAGGA | 1542

Val94Lys AAAAGACGAGTTCCTTTACCACCACCTAGAATGACAGAGGCAACA

GTT-AAA GCITTTGGATCTGCCGTTGGTTGTTCCTCAA

TGGTGGTAAAGGAACTC 1543 1112/01 01:47 pm 03137.009 — {NY}793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration

IH GAGTTCCTTTACCACCA 1544 ncreased Starch CAAGCAGAAGAGCTAAACCAGCTGTTCCTATIGGTGGTTGTTAC 545

ADPGPP GGCTAATTGATGTACAAATGAGTAACTGCATTAACAGTGGCATAC

Lycopersicon GGAAAATTTTCATCTTAACACAGTTCAATTC esculentum GAATTGAACTGTGTTAAGATGAAAATTTTCCGTATGCCACTGTTAA | 1546

Pro122Leu TGCAGTTACTCATTIGTACATCAATTAGCCGGTAACAACCACCAAT

CCA-CAA AGGAACAGCTGGTTTAGCTCTTCTGCTTG

TGATGTACAAATGAGTA 1547

TACTCATTIGTACATCA 1548 [ Increased Starch CACAGTTCAATTCCTTTTCCCTCAATCGTCACCTTGCCCGCACGTA | 1549

ADPGPP TAATTTTGGAAATAATGTGGGTTTTGGAGATGGATTTGTGGAGGTT

Lycopersicon TTAGCTGCAACCCAGACTCCAGGGGATGC esculentum GCATCCCCTGGAGTCTGGGTTGCAGCTAAAACCTCCACAAATCCA | 1550

Gly158Asn TCTCCAAAACCCACATTATTTCCAAAATTATACGTGCGGGCAAGGT

GGA-AAT GACGATTGAGGGAAAAGGAATTGAACTGTG

TGGAAATAATGTGGGTT 1551

AACCCACATTATTTCCA

Increased Starch ACAGTTCAATTCCTTTTCCCTCAATCGTCACCTTGCCCGCACGTA 563

ADPGPP TAATTTTGGAAATAACGTGGGTTTTGGAGATGGATTTGTGGAGGT

Lycopersicon TTTAGCTGCAACCCAGACTCCAGGGGATGC esculentum GCATCCCCTGGAGTCTGGGTTGCAGCTAAAACCTCCACAAATCCA | 1554

Gly158Asn TCTCCAAAACCCACGTTATTTCCAAAATTATACGTGCGGGCAAGGT

GGA-AAC GACGATTGAGGGAAAAGGAATTGAACTGTG

PS TGGAAATAACGTGGGTT 1555

AACCCACGTTATTTCCA 1556

Increased Starch ACGTAGATTTGGAAAAAAGAGACCCAAGTACAGTTGTAGCAATTAT 557

ADPGPP ACTAGGTGGAGGTAAAGGAACTCGTCTCTTCCCTCTCACCAAGCG

Cicer arietinum ACGAGCCAAGCCTGCTGTTCCAATTGGAGG

Ala101Lys CCTCCAATTGGAACAGCAGGCTTGGCTCGTCGCTTGGTGAGAGG | 1558

GCT-AAA GAAGAGACGAGTTCCITTACCTCCACCTAGTATAATTGCTACAACT

GTACTTGGGTCTCTTTTTTCCAAATCTACGT

TGGAGGTAAAGGAACTC 1659

GAGTTCCTITTACCTCCA 1560 1112/01 01:47 pm 03137.009 — [NY]793550.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ) Alteration increased Starch CAAGCGACGAGCCAAGCCTGCTGITCCAATTGGAGGTGCTTATA | 156 : ADPGPP GGCTGATAGATGTACTAATGAGTAACTGCATCAATAGTGGGATCA

Cicer arietinum ACAAAGTATACATTCTCACTCAATTTAATTC

Pro129Leu GAATTAAATTGAGTGAGAATGTATACTTTGTTGATCCCACTATTGA | 1562

CCA-CTA TGCAGTTACTCATTAGTACATCTATCAGCCTATAAGCACCTCCAAT

TGGAACAGCAGGCTTGGCTCGTCGCTTGG

AGATGTACTAATGAGTA

TACTCATTAGTACATCT _ 1564

Increased Starch CTCAATTTAATTCAGCCTCACTCAACAGGCATATTGCACGTGCTTA | 1565

ADPGPP TAACTCTGGTACTAATGTCACTTTTGGAGATGGCTATGTTGAGGTT

® Cicer arietinum CTTGCAGCAACTCAAACTCCAGGGGAGCA

Gly165Asn TGCTCCCCTGGAGTTTGAGTTGCTGCAAGAACCTCAACATAGCCA | 1566

GGA-AAT TCTCCAAAAGTGACATTAGTACCAGAGTTATAAGCACGTGCAATAT

GCCTGTTGAGTGAGGCTGAATTAAATTGAG

TGGTACTAATGTCACTT

AAGTGACATTAGTACCA 1568

ADPGPP TAACTCTGGTACTAACGTCACTTTTGGAGATGGCTATGTTGAGGTT

Cicer arietinum CTTGCAGCAACTCAAACTCCAGGGGAGCA

Gly165Asn TGCTCCCCTGGAGTTTGAGTTGCTGCAAGAACCTCAACATAGCCA | 1570

GGA-AAC TCTCCAAAAGTGACGTTAGTACCAGAGTTATAAGCACGTGCAATAT

GCCTGTTGAGTGAGGCTGAATTAAATTGAG

TGGTACTAACGTCACTT 1571

AAGTGACGTTAGTACCA

® ADPGPP ATACTGCCAGGCGGTAAAGGGACACACCTATTCCCTCTCACCAAT

Ipomoea batatas CGAGCTGCAACCCCTGCTGTTCCACTTGGAG

Ala%4Lys CTCCAAGTGGAACAGCAGGGGTTGCAGCTCGATTGGTGAGAGGG | 1574

GCA-AAA AATAGGTGTGTCCCTITACCGCCTGGCAGTATGATTGCAGCCACA

TTCTTAGGGTTTGCCCGACGCCTCTCCAATAT

CAGGCGGTAAAGGGACA 1575

TGTCCCTITACCGCCTG 1576 ncreased Starch AATCGAGCTGCAA CTGITCCACTTGGAGGATGCTATA | 1577

ADPGPP GGTTGATCGACATTCTAATGAGCAACTGCATCAACAGCGGGGTTA

Ipomoea batatas ACAAGATCTTTGTGCTGACCCAGTTCAATTC

Pro122Leu GAATTGAACTGGGTCAGCACAAAGATCTTGTTAACCCCGCTGTTG | 1578 "25 CCA-CTA ATGCAGTTGCTCATTAGAATGTCGATCAACCTATAGCATCCTCCAA

GTGGAACAGCAGGGGTTGCAGCTCGATTGG

CGACATTCTAATGAGCA 1579 11/12/01 01:47 pm 03137.009 — [NY)793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration 1TGCTCATTAGAATGTCG | 1580

Increased Starch TGACCCAGTTCAATTCAGCTTCTCTTAACCGTCACATTTCCCGTAC | 1581

ADPGPP CGTCTTTGGCAATAATGTGAGCTTCGGAGATGGATTTGTTGAGGT

Ipomoea batatas GCTGGCTGCAACCCAAACACAAGGGGAAAC

Gly157Asn GTTTCCCCTTGTGTTTGGGTTGCAGCCAGCACCTCAACAAATCCA | 1582

GGT-AAT TCTCCGAAGCTCACATTATTGCCAAAGACGGTACGGGAAATGTGA

CGGTTAAGAGAAGCTGAATTGAACTGGGTCA

TGGCAATAATGTGAGCT 1583

AGCTCACATTATTGCCA 1584

Increased Starch TGACCCAGTTCAATTCAGCTTCTCTTAACCGTCACATTTCCCGTAC | 1585 @® ADPGPP CGTCTTTGGCAATAACGTGAGCTTCGGAGATGGATTTGTTGAGGT

Ipomoea batatas GCTGGCTGCAACCCAAACACAAGGGGAAAC

Gly157Asn GTTTCCCCTTGTGTTTGGGTTGCAGCCAGCACCTCAACAAATCCA | 1586

GGT-AAC TCTCCGAAGCTCACGTTATTGCCAAAGACGGTACGGGAAATGTGA

CGGTTAAGAGAAGCTGAATTGAACTGGGTCA

TGGCAATAACGTGAGCT 1587

AGCTCACGTTATTGCCA

Increased Starch CATTCCGGAGGAACTTTGCGGATCCAAATGAGGTTGCTGCTGTTA | 1589

ADPGPP TATTGGGTGGTGGCAAAGGGACTCAACTTTTTCCTCTCACAAGCA

Oryza sativa CAAGGGCCACGCCTGCTGTTCCTATTGGAGG

Thr36Lys CCTCCAATAGGAACAGCAGGCGTGGCCCTTGTGCTTGTGAGAGG | 1590

ACC-AAA AAAAAGTTGAGTCCCTTTGCCACCACCCAATATAACAGCAGCAAC

CTCATTTGGATCCGCAAAGTTCCTCCGGAATG

TGGTGGCAAAGGGACTC 1591

PY GAGTCCCITTGCCACCA ncreased Starc AAGCACAAGGGCCACGCCTGCTGTTCCTATTGGAGGATGCTATA 93

ADPGPP GGCTTATCGATATCCTCATGAGCAACTGTTTCAACAGTGGCATAAA

Oryza sativa CAAGATATTCATAATGACTCAATTCAACTC

Pro124Leu GAGTTGAATTGAGTCATTATGAATATCTTGTTTATGCCACTGTTGA | 1594

CCC-CTC AACAGTTGCTCATGAGGATATCGATAAGCCTATAGCATCCTCCAAT

AGGAACAGCAGGCGTGGCCCTTGTGCTTG

CGATATCCICATGAGCA 1595

TGCTCATGAGGATATCG 1596

Increased Starch TGACTCAATTCAACTCAGCATCTCTTAATCGTCACATTCATCGTAC | 1597

ADPGPP GTACCTTGGTGGTAATATCAACTTTACTGATGGTTCTGTTGAGGTA

Oryza sativa TTAGCCGCTACACAAATGCCTGGGGAGGC

Gly159Asn GCCTCCCCAGGCATTTGTGTAGCGGCTAATACCTCAACAGAACCA | 1598

GGA-AAT TCAGTAAAGTTGATATTACCACCAAGGTACGTACGATGAATGTGA

CGATTAAGAGATGCTGAGTTGAATTGAGTCA

11712001 01:47 pm 03137.009 — [N)793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ’ Alteration

TGGTGGTAATATCAACT 1599

AGTTGATATTACCACCA 1600 ncreased Starc TGACTCAATTCAACTCAGCATCTCTTAATCGTCACATTCATCGTAC

ADPGPP GTACCTTGGTGGTAACATCAACTTTACTGATGGTTCTGTTGAGGTA

Oryza sativa TTAGCCGCTACACAAATGCCTGGGGAGGC

Gly159Asn GCCTCCCCAGGCATTTGTGTAGCGGCTAATACCTCAACAGAACCA | 1602

GGA-AAC TCAGTAAAGTTGATGTTACCACCAAGGTACGTACGATGAATGTGA

CGATTAAGAGATGCTGAGTTGAATTGAGTCA

TGGTGGTAACATCAACT 1603

AGTTGATGTTACCACCA

®

ADPGPP ATACTCGGCGGCGGCAAAGGGACTCAGCTCTTCCCACTCACGAG

Triticum aestivum CACAAGGGCCACACCTGCTGTTCCTATTGGAGG

Thr80Lys CCTCCAATAGGAACAGCAGGTGTGGCCCTTGTGCTCGTGAGTGG | 1606

ACC-AAA GAAGAGCTGAGTCCCTTTGCCGCCGCCGAGTATGACGGCCGCAA

CCTCGTTCGGATCGCTTAATCCTCCTGAAGGAC

CGGCGGCAAAGGGACTC

GAGTCCCITTGCCGCCG 1608

ADPGPP GGCTCATCGACATTCICATGAGCAACTGCTTCAACAGTGGCATCA

Triticum aestivum ACAAGATATTCGTCATGACCCAGTTCAACTC

Pro108Leu GAGTTGAACTGGGTCATGACGAATATCTTGTTGATGCCACTGTTG | 1610

CCC-CTC AAGCAGTTGCTCATGAGAATGTCGATGAGCCTGTAACATCCTCCA

ATAGGAACAGCAGGTGTGGCCCTTGTGCTCG

CGACATTCTCATGAGCA 1611 ® TGCTCATGAGAATGTCG 1612

Increased Starch TGACCCAGTTCAACTCGGCCTCCCTTAATCGTCACATTCACCGCA | 1613

ADPGPP CCTACCTCGGCGGGAATATCAATTTCACTGATGGATCCGTTGAGG

Triticum aestivum TATTGGCCGCGACGCAAATGCCCGGGGAGGC

Gly143Asn GCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACGGATCC | 1614

GGA-AAT ATCAGTGAAATTGATATTCCCGCCGAGGTAGGTGCGGTGAATGTG

ACGATTAAGGGAGGCCGAGTTGAACTGGGTCA

CGGCGGGAATATCAATT 1615

AATTGATATTCCCGCCG 1616 : ADPGPP CCTACCTCGGCGGGAACATCAATTTCACTGATGGATCCGTTGAGG iticum aestivum TATTGGCCGCGACGCAAATGCCCGGGGAGGC

Gly143Asn

GGA-AAC 111201 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

GCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACGGATCC | 1618 ! ATCAGTGAAATTGATGTTCCCGCCGAGGTAGGTGCGGTGAATGTG

ACGATTAAGGGAGGCCGAGTTGAACTGGGTCA

CGGCGGGAACATCAATT 1619

AATTGATGTTCCCGCCG 1620

ADPGPP TTTTGGGTGGAGGCAAAGGAGTTCAACTCTTTCCTCTGACAAGCA

Oryza sativa CAAGGGCTACCCCCGCTGTTCCTGTTGGAGG

Thr95Lys CCTCCAACAGGAACAGCGGGGGTAGCCCTTGTGCTTGTCAGAGG | 1622

ACT-AAA AAAGAGTTGAACTCCITTGCCTCCACCCAAAATGACAGCAGAAAC ® GTGGCTTGCATCAGCATAATTCTTTCGGGAGG

TGGAGGCAAAGGAGTTC 1623

GAACTCCTTTGCCTCCA 1624

Tnoreased Starch | CAAGCAGAAGGGCTACCCCCGCTGTTCCIGTTGGAGGATGTTACA | 1625

ADPGPP GGCTTATTGACATCCITATGAGCAATTGCTTCAATAGCGGAATAAA

Oryza sativa TAAAATATTTGTGATGACTCAGTTCAATTC

Pro123Leu GAATTGAACTGAGTCATCACAAATATTTTATTTATTCCGCTATTGAA | 1626

CCT-CTT GCAATTGCTCATAAGGATGTCAATAAGCCTGTAACATCCTCCAACA

GGAACAGCGGGGGTAGCCCTTGTGCTTG

TGACATCCITATGAGCA 1627

TGCTCATAAGGATGTCA

Increased Starch TGACTCAGTTCAATTCTGCTTCTCTTAATCGCCATATCCATCATACA| 1629

ADPGPP TACCTTGGTGGGAATATCAACTTTACTGATGGGTCTGTGCAGGTA

Oryza sativa TTGGCTGCTACACAAATGCCTGACGAACC

Gly158Asn GGTTCGTCAGGCATTTGTGTAGCAGCCAATACCTGCACAGACCCA | 1630 ® GGG-AAT TCAGTAAAGTTGATATTCCCACCAAGGTATGTATGATGGATATGGC

GATTAAGAGAAGCAGAATTGAACTGAGTCA

TGGTGGGAATATCAACT 1631

AGTTGATATTCCCACCA 1632

ADPGPP TACCTTGGTGGGAACATCAACTTTACTGATGGGTCTGTGCAGGTA

Oryza sativa TTGGCTGCTACACAAATGCCTGACGAACC

Gly158Asn GGTTCGTCAGGCATTTGTGTAGCAGCCAATACCTGCACAGACCCA | 1634

GGG-AAC TCAGTAAAGTTGATGITCCCACCAAGGTATGTATGATGGATATGG

CGATTAAGAGAAGCAGAATTGAACTGAGTCA

TGGTGGGAACATCAACT 1635

AGTTGATGTTCCCACCA 1636 1112/01 01:47 pm 03137.003 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ) Alteration

ADPGPP ATACTCGGCGGTGGCAAAGGGACTCAGCTCTTCCCTCTCACAAG

Triticum aestivum CACAAGGGCCACACCTGCTGTTCCTATTGGAGG

Thr99Lys CCTCCAATAGGAACAGCAGGTGTGGCCCTTGTGCTTGTGAGAGG | 1638

ACC-AAA GAAGAGCTGAGTCCCITTGCCACCGCCGAGTATGACGGCCGCGA

CCTCGTTCGGATCGGCGTAATTCCTGCGGAAGG

CGGTGGCAAAGGGACTC 1639

GAGTCCCITTGCCACCG 1640 oreased Starch | CAAGCACAAGGGCCACACCTGCTGTTCOTATT GGAGGATGTTAGA | 1641

ADPGPP GGCTCATCGATATTCTICATGAGCAACTGCTTCAATAGTGGCATCAA

® Triticum aestivum CAAGATATTCGTCATGACGCAGTTCAACTC

Pro127Leu GAGTTGAACTGCGTCATGACGAATATCTTGTTGATGCCACTATTGA | 1642

CCC-CTC AGCAGTTGCTCATGAGAATATCGATGAGCCTGTAACATCCTCCAA

TAGGAACAGCAGGTGTGGCCCTTGTGCTTG

CGATATTCICATGAGCA

TGCTCATGAGAATATCG | 1644

Increased Starch TGACGCAGTTCAACTCGGCCTCTCTTAATCGTCACATTCACCGCA | 1645

ADPGPP CCTACCTCGGCGGGAATATCAATTTCACTGATGGATCTGTTGAGG

Triticum aestivum TATTGGCCGCGACGCAAATGCCCGGGGAGGC

Gly162Asn GCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACAGATCC 1646

GGA-AAT ATCAGTGAAATTGATATTICCCGCCGAGGTAGGTGCGGTGAATGTG

ACGATTAAGAGAGGCCGAGTTGAACTGCGTCA

CGGCGGGAATATCAATT

AATTGATATTICCCGCCG

® [Tncreased Stare ACGCAGTTCAACTC CTCTCTTAATCGTCACATTCACCGCA | 1649

ADPGPP CCTACCTCGGCGGGAACATCAATTTCACTGATGGATCTGTTGAGG

Triticum aestivum TATTGGCCGCGACGCAAATGCCCGGGGAGGC

Gly162Asn GCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACAGATCC 1650

GGA-AAC ATCAGTGAAATTGATGTTCCCGCCGAGGTAGGTGCGGTGAATGTG

ACGATTAAGAGAGGCCGAGTTGAACTGCGTCA

CGGCGGGAACATCAATT 1651

AATTGATGTTCCCGCCG 1652

ADPGPP TTTTGGGTGGTGGTAAAGGGACTCAGCTTTTCCCTCTCACAAGCA

Zea mays CAAGGGCCACCCCTGCTGTTCCTATTGGAGG

Thr96Lys CCTCCAATAGGAACAGCAGGGGTGGCCCTTGTGCTTGTGAGAGG | 1654

ACC-AAA GAAAAGCTGAGTCCCTTTACCACCACCCAAAATGACGGCAGCGAC

TTCATTAGGATCAGCATAATTCCTCCGAAAAG

TGGTGGTAAAGGGACTC 1655 11/12/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET i2-2001 Uivog: oi!

Phenotype, Gene,

Plant & Targeted Altering Oligos SEQ : D . \ NO:

Alteration

TT] GAGTCCCTTTACCACCA 1656

Increased Starch CAAGCACAA CACCCCTGCT ATTGGAGGATGTTACA 657

ADPGPP GGCTTATTGATATCCTCATGAGCAACTGTTTCAACAGTGGCATAAA

Zea mays CAAGATATTTGTTATGACTCAGTTCAACTC

Pro124Leu GAGTTGAACTGAGTCATAACAAATATCTTGTTTATGCCACTGTTGA | 1658

CCC-CTC AACAGTTGCTCATGAGGATATCAATAAGCCTGTAACATCCTCCAAT

AGGAACAGCAGGGGTGGCCCTTGTGCTTG

TGATATCCICATGAGCA 1659

TGCTCATGAGGATATCA 1660 » Increased Starch TGACTCAGTTCAACTCAGCTTCTCTTAACCGTCACATTCATCGTAC | 1661

ADPGPP CTATCTTGGTGGGAATATCAACTTCACTGATGGATCTGTTGAGGT

Zea mays GCTGGCTGCAACACAAATGCCTGGGGAGGC

Gly159Asn GCCTCCCCAGGCATTTGTGTTGCAGCCAGCACCTCAACAGATCCA | 1662

GGG-AAT TCAGTGAAGTTGATATTCCCACCAAGATAGGTACGATGAATGTGA

CGGTTAAGAGAAGCTGAGTTGAACTGAGTCA

TGGTGGGAATATCAACT

AGTTGATATICCCACCA ncreased Starch ACTCAGTTCAACTCAGCTTCTCTTAACCGTCACATICA AC | 1665

ADPGPP CTATCTTGGTGGGAACATCAACTTCACTGATGGATCTGTTGAGGT

Zea mays GCTGGCTGCAACACAAATGCCTGGGGAGGC

Gly159Asn GCCTCCCCAGGCATTTGTGTTGCAGCCAGCACCTCAACAGATCCA | 1666

GGG-AAC TCAGTGAAGTTGATGTTCCCACCAAGATAGGTACGATGAATGTGA

CGGTTAAGAGAAGCTGAGTTGAACTGAGTCA

TGGTGGGAACATCAACT 1667 ® AGTTGATGTTCCCACCA 1668

Increased Starc CTTGAGA AAAAGAAGGGCGATGCAAGGACAGTAGTAGCAAT 0

ADPGPP CATTCTAGGAGGGGGAAAGGGAACTCGTCTTTTCCCCCTCACCAA

Solanum tuberosum | ACGTCGTGCTAAGCCTGCCGTTCCAATGGGAG

Ala58Lys CTCCCATTGGAACGGCAGGCTTAGCACGACGTTTGGTGAGGGGG | 1670

GCG-AAG AAAAGACGAGTTCCCTTTCCCCCTCCTAGAATGATTGCTACTACTG

TCCTTGCATCGCCCTTCTTTTGCCTCTCAAG

GAGGGGGAAAGGGAACT 1671

AGTTCCCTITCCCCCTC 1672 ) Increased Starc CCAAACGTCGTGCTAAGCCTGCCGTTCCAATGGGAGGAGCATATA | 1673

ADPGPP GGCTAATTGATGTACTAATGAGCAACTGTATTAACAGTGGCATCAA

Solanum tuberosum | CAAAGTATACATTCTCACTCAATTCAACTC

Pra86leu GAGTTGAATTGAGTGAGAATGTATACTTTGTTGATGCCACTGTTAA | 1674

CCA-CTA TACAGTTGCTCATTAGTACATCAATTAGCCTATATGCTCCTCCCAT

TGGAACGGCAGGCTTAGCACGACGTTTIGG

1411201 01:47 pm 03137.009 — (NY}793558.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ; Alteration

TGATGTACTAATGAGCA 1675 ‘ TGCTCATTAGTACATCA 1676 ncreased Starc TCAATTCAACTCAGCCTCACTTAACAGGCATATAGCTCGTGCTTA 677

ADPGPP CAACTTTGGCAATAATGTCACATTCGAGAGTGGCTATGTCGAGGT

Solanum tuberosum | CTTAGCAGCAACTCAAACACCAGGTGAATT

Gly122Asn AATTCACCTGGTGTTTGAGTTGCTGCTAAGACCTCGACATAGCCA | 1678

GGG-AAT CTCTCGAATGTGACATTATTGCCAAAGTTGTAAGCACGAGCTATAT

GCCTGTTAAGTGAGGCTGAGTTGAATTGAG

TGGCAATAATGTCACAT 1679 ® ATGTGACATTATTGCCA 1680

Increased Starc CAATTCAACTCA TCACTTAACAGGCATATAGCT: CTTA | 1681

ADPGPP CAACTTTGGCAATAACGTCACATTCGAGAGTGGCTATGTCGAGGT

Solanum tuberosum | CTTAGCAGCAACTCAAACACCAGGTGAATT

Gly122Asn AATTCACCTGGTGTTTGAGTTGCTGCTAAGACCTCGACATAGCCA | 1682

GGG-AAC CTCTCGAATGTGACGTTATTGCCAAAGTTGTAAGCACGAGCTATAT

GCCTGTTAAGTGAGGCTGAGTTGAATTGAG

TGGCAATAACGTCACAT 1683

ATGTGACGTTATTGCCA 1684 ncreased Starc TATTTGAATCTCCAAAAGCTGACCCAAAAAATGTGGCTGCAATTGT 68

ADPGPP GCTGGGTGGTGGTAAAGGGACTCGCCTCTTTCCTCTTACTAGCAG

Beta vulgaris GAGAGCTAAGCCAGCAGTGCCAATTGGAGG

AlaS8Lys CCTCCAATTGGCACTGCTGGCTTAGCTCTCCTGCTAGTAAGAGGA | 1686

GCT-AAA AAGAGGCGAGTCCCTTTACCACCACCCAGCACAATTGCAGCCACA

TTTTTTGGGTCAGCTTTTGGAGATTCAAATA

® TGGTGGTAAAGGGACTC 1687

GAGTCCCTTTACCACCA 1688

Increased Starc TATTTGAATCTCCAAAAGCTGACCCAAAAAATGTC CAATTGT [ 1689

ADPGPP GCTGGGTGGTGGTAACGGGACTCGCCTCTTTCCTCTTACTAGCAG

Beta vulgaris GAGAGCTAAGCCAGCAGTGCCAATTGGAGG

Ala98Lys CCTCCAATTGGCACTGCTGGCTTAGCTCTCCTGCTAGTAAGAGGA | 1690

GCT-AAC AAGAGGCGAGTCCCGTTACCACCACCCAGCACAATTGCAGCCAC

ATTTTTTGGGTCAGCTTTTGGAGATTCAAATA

TGGTGGTAACGGGACTC 1691 . GAGTCCCGTTACCACCA 1692

Increased Starch TAGCAGGAGAGCTAAGCCAGCAGTGCCAATTGGAGGGTGTTAC 6

ADPGPP AGGCTGATTGATGTGCTTATGAGCAACTGCATCAACAGTGGCATT

Bata vulgar AGAAAGATTTTCATTCTTACCCAGTTCAATTC

Pro126Leu

CCT-CTT 141201 01:47pm 03137.009 — [NY}793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

GAATTGAACTGGGTAAGAATGAAAATCTTTCTAATGCCACTGTTGA | 1694

TGCAGTTGCTCATAAGCACATCAATCAGCCTGTAACACCCTCCAA

TTGGCACTGCTGGCTTAGCTCTCCTGCTAG

TGATGTGCTTATGAGCA : 1695

TGCTCATAAGCACATCA 1696 ncreased Starc AGTTCAATTCGTTTTCGCTTAATCGTCATCTTGCTCGAACCTA | 1697

ADPGPP TAATTTTGGAGATAATGTGAATTTTGGGGATGGCTTTGTGGAGGTT

Beta vulgaris TTTGCTGCTACACAAACACCTGGAGAATC

Gly162Asn GATTCTCCAGGTGTTTGTGTAGCAGCAAAAACCTCCACAAAGCCA | 1698

GGT-AAT TCCCCAAAATTCACATTATCTCCAAAATTATAGGTTCGAGCAAGAT ® GACGATTAAGCGAAAACGAATTGAACTGGG

TGGAGATAATGTGAATT 1699

AATTCACATTATCTCCA 1700 ncreased Starch AGTTCAATTCGTTTTCGCTTAATCGTCATCTTGCTCGAACCTA 701

ADPGPP TAATTTTGGAGATAACGTGAATTTTGGGGATGGCTTTGTGGAGGT

Beta vulgaris TTTTGCTGCTACACAAACACCTGGAGAATC

Gly162Asn GATTCTCCAGGTGTTTGTGTAGCAGCAAAAACCTCCACAAAGCCA | 1702

GGT-AAC TCCCCAAAATTCACGTTATCTCCAAAATTATAGGTTCGAGCAAGAT

GACGATTAAGCGAAAACGAATTGAACTGGG

TGGAGATAACGTGAATT 1703

AATTCACGTTATCTCCA

14/12/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Table 21

Oligonucleotides to produce plants with waxy starch

Phenotype, Gene, , Plant & Targeted Altering Oligos

Alteration

Waxy starch GAATCCAGGTAAACGGGTAGTTCATAATGGCAACTGTGACTGCTT | 1705

GBSS CTTCTAACTTTGTGTGAAGAACTTCACTTTTCAACAATCATGGTGCT

Arabidopsis thaliana {TCTTCATGCTCTGATGTCGCTCAGATTAC

Ser12Term GTAATCTGAGCGACATCAGAGCATGAAGAAGCACCATGATTGTTG | 1706

TCA-TGA AAAAGTGAAGTTCTTCACACAAAGTTAGAAGAAGCAGTCACAGTTG

CCATTATGAACTACCCGTTTACCTGGATTC

CTTTGTGTGAAGAACTT

® AAGTTCTTCACACAAAG 1708

Waxy starch ATCCAGGTAAACGGGTAGTTCATAATGGCAACTGTGACTGCTTCTT | 1709

GBSS CTAACTTTGTGTCATGAACTTCACTTTTCAACAATCATGGTGCTTCT

Arabidopsis thaliana |TCATGCTCTGATGTCGCTCAGATTACCT

Arg13Term AGGTAATCTGAGCGACATCAGAGCATGAAGAAGCACCATGATTGT | 1710 16 AGA-TGA TGAAAAGTGAAGTTCATGACACAAAGTTAGAAGAAGCAGTCACAGT

TGCCATTATGAACTACCCGTTTACCTGGAT

TTGTGTCATGAACTTCA

TGAAGTTCATGACACAA 1712

Waxy starch TAAACGGGTAGTTCATAATGGCAACTGTGACTGCTTCTTCTAACTT | 1713

GBSS TGTGTCAAGAACTTGACTTTTCAACAATCATGGTGCTTCTTCATGCT

Arabidopsis thaliana |CTGATGTCGCTCAGATTACCTTAAAAGG

Ser15Term CCTTTTAAGGTAATCTGAGCGACATCAGAGCATGAAGAAGCACCAT | 1714

TCA-TGA GATTGTTGAAAAGTCAAGTTCTTGACACAAAGTTAGAAGAAGCAGT ® CACAGTTGCCATTATGAACTACCCGTTTA

AAGAACTTGACTTTTCA 1715

TGAAAAGTCAAGTTCTT 1716

Waxy starch TGACTGCTTCTTCTAACTTTGTGTCAAGAACTTCACTTTTCAACAAT | 1717

GBSS CATGGTGCTTCTTGATGCTCTGATGTCGCTCAGATTACCTTAAAAG

Arabidopsis thaliana |GCCAATCCTTGACTCATTGTGGGTTAAG

Ser24Term CTTAACCCACAATGAGTCAAGGATTGGCCTTTTAAGGTAATCTGAG | 1718

TCA-TGA CGACATCAGAGCATCAAGAAGCACCATGATTGTTGAAAAGTGAAG

TTCTTGACACAAAGTTAGAAGAAGCAGTCA

: TGCTTCTTGATGCTCTG 1719

CAGAGCATCAAGAAGCA 1720 11/12/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Waxy starch TGCTTCTTCTAACTTTGTGTCAAGAACTTCACTTTTCAACAATCATG | 1721

GBSS GTGCTTCTTCATGATCTGATGTCGCTCAGATTACCTTAAAAGGCCA

) Arabidopsis thaliana |ATCCTTGACTCATTGTGGGTTAAGGTCA

Cys25Term TGACCTTAACCCACAATGAGTCAAGGATTGGCCTTTTAAGGTAATC | 1722

TGC-TGA TGAGCGACATCAGATCATGAAGAAGCACCATGATTGTTGAAAAGT

GAAGTTCTTGACACAAAGTTAGAAGAAGCA

TCTTCATGATCTGATGT 1723

ACATCAGATCATGAAGA

Waxy starch GTAACAGCTTCACAGTTGGTGTCACATGTCCATGGTGGAGCAACG | 1725

GBSS TCTTCACCGGATACTTAAACAAACTTGGCCCAGGTTGGCCTCAGG

® Antirrhinum majus AACCAGCAATTCACTCACAATGGGTTGAGAT

Lys24Term ATCTCAACCCATTGTGAGTGAATTGCTGGTTCCTGAGGCCAACCTG| 1726

AAA-TAA GGCCAAGTTTGTTTAAGTATCCGGTGAAGACGTTGCTCCACCATG

GACATGTGACACCAACTGTGAAGCTGTTAC

CGGATACTTAAACAAAC 1727

GTTTGTTTAAGTATCCG 1728

Waxy starch CACAGTTGGTGTCACATGTCCATGGTGGAGCAACGTCTTCACCGG | 1729

GBSS ATACTAAAACAAACTAGGCCCAGGTTGGCCTCAGGAACCAGCAAT

Antirrhinum majus TCACTCACAATGGGTTGAGATCAATAAACAT

Leu27Term ATGTTTATTGATCTCAACCCATTGTGAGTGAATTGCTGGTTCCTGA | 1730

TTG-TAG GGCCAACCTGGGCCTAGTTTGTTTTAGTATCCGGTGAAGACGTTG

CTCCACCATGGACATGTGACACCAACTGTG

AACAAACTAGGCCCAGG 1731

CCTGGGCCTAGTTTGTT 1732 ® Waxy starch TTGGTGTCACATGTCCATGGTGGAGCAACGTCTTCACCGGATACT | 1733

GBSS AAAACAAACTTGGCCTAGGTTGGCCTCAGGAACCAGCAATTCACT

Antirrhinum majus CACAATGGGTTGAGATCAATAAACATGGTTG

GIn29Term CAACCATGTTTATTGATCTCAACCCATTGTGAGTGAATTGCTGGTT 1734

CAG-TAG CCTGAGGCCAACCTAGGCCAAGTTTGTTTTAGTATCCGGTGAAGA

CGTTGCTCCACCATGGACATGTGACACCAA

ACTTGGCCTAGGTTGGC 1735

GCCAACCTAGGCCAAGT 1736

Waxy starch GGTGGAGCAACGTCTTCACCGGATACTAAAACAAACTTGGCCCAG | 1737

GBSS GTTGGCCTCAGGAACTAGCAATTCACTCACAATGGGTTGAGATCA i Antirrhinum majus ATAAACATGGTTGATAAGCTTCAAATGAGGA

GIn35Term TCCTCATTTGAAGCTTATCAACCATGTTTATTGATCTCAACCCATTG | 1738

CAG-TAG TGAGTGAATTGCTAGTTCCTGAGGCCAACCTGGGCCAAGTTTGTTT

TAGTATCCGGTGAAGACGTTGCTCCACC

1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ) Alteration

TCAGGAACTAGCAATTC 1739

GAATTGCTAGTTCCTGA 1740

Waxy starch GGAGCAACGTCTTCACCGGATACTAAAACAAACTTGGCCCAGGTT | 1741

GBSS GGCCTCAGGAACCAGTAATTCACTCACAATGGGTTGAGATCAATAA

Antirrhinum majus ACATGGTTGATAAGCTTCAAATGAGGAACA

GIn36Term TGTTCCTCATTTGAAGCTTATCAACCATGTTTATTGATCTCAACCCA | 1742

CAA-TAA TTGTGAGTGAATTACTGGTTCCTGAGGCCAACCTGGGCCAAGTTT

GTTTTAGTATCCGGTGAAGACGTTGCTCC

GGAACCAGTAATTCACT 1743 ® AGTGAATTACTGGTTCC

Waxy starch GTGATGGCGACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTG | 1745

GBSS GGGGTGCCACTTCTIGAGAATCAAAAGTGGGGTTGGGTCAATTAG

Ipomoea batatas CCCTGAGGAGCCAAGCTGTGACTCACAATG

Gly20Term CATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATTGACCCAACC | 1746

GGA-TGA CCACTTTTGATTCTCAAGAAGTGGCACCCCCACAGACATGAGAAA

CAAAGTGTGAGGCAGTTATAGTCGCCATCAC

CCACTTCTIGAGAATCA

TGATTCTCAAGAAGTGG 1748

Waxy starch ATGGCGACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTGGGG | 1749

GBSS GTGCCACTTCTGGATAATCAAAAGTGGGGTTGGGTCAATTAGCCC

Ipomoea batatas TGAGGAGCCAAGCTGTGACTCACAATGGGT

Glu21Term ACCCATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATTGACCCA | 1750

GAA-TAA ACCCCACTTTTGATTATCCAGAAGTGGCACCCCCACAGACATGAG ® AAACAAAGTGTGAGGCAGTTATAGTCGCCAT

CTTCTGGATAATCAAAA 1751

TTTTGATTATCCAGAAG

Waxy starch CGACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTGGGGGTGC | 1753

GBSS CACTTCTGGAGAATGAAAAGTGGGGTTGGGTCAATTAGCCCTGAG

Ipomoea batatas GAGCCAAGCTGTGACTCACAATGGGTTGAG

Ser22Term CTCAACCCATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATTGA | 1754

TCA-TGA CCCAACCCCACTTTTCATTCTCCAGAAGTGGCACCCCCACAGACAT

GAGAAACAAAGTGTGAGGCAGTTATAGTCG

. TGGAGAATGAAAAGTGG 1755

CCACTTTTCATTCTCCA 1756 1142/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos .

Waxy starch ACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTGGGGGTGCCA | 1757 } GBSS CTTCTGGAGAATCATAAGTGGGGTTGGGTCAATTAGCCCTGAGGA

Ipomoea batatas GCCAAGCTGTGACTCACAATGGGTTGAGAC

Lys23Term GTCTCAACCCATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATT | 1758

AAA-TAA GACCCAACCCCACTTATGATTCTCCAGAAGTGGCACCCCCACAGA

CATGAGAAACAAAGTGTGAGGCAGTTATAGT

GAGAATCATAAGTGGGG 1759

CCCCACTTATGATTCTC 1760

Waxy starch CCTCACACTTTGTTTCTCATGTCTGTGGGGGTGCCACTTCTGGAGA| 1761

PY GBSS ATCAAAAGTGGGGTAGGGTCAATTAGCCCTGAGGAGCCAAGCTGT

Ipomoea batatas GACTCACAATGGGTTGAGACCTGTGAACAA

Leu26Term TTGTTCACAGGTCTCAACCCATTGTGAGTCACAGCTTGGCTCCTCA | 1762

TTG-TAG GGGCTAATTGACCCTACCCCACTTTTGATTCTCCAGAAGTGGCACC

CCCACAGACATGAGAAACAAAGTGTGAGG

AGTGGGGTAGGGTCAAT 1763

ATTGACCCTACCCCACT 1764

Waxy starch CATCGGCGATTGTTGCTCCTTACTGCTCTCTCACAGAATGGCAACG]| 1765

GBSS GTGACGGGGTCTTAGGTGGTGTCGAGAAGCGCGTGCTTCAATTCC

Astragalus CAGGGAAGAACAGAAGCCAAAGTGAATTCA membranaeus TGAATTCACTTTGGCTTCTGTTCTTCCCTGGGAATTGAAGCACGCG | 1766

Tyr8Term CTTCTCGACACCACCTAAGACCCCGTCACCGTTGCCATTCTGTGA

TAT-TAG GAGAGCAGTAAGGAGCAACAATCGCCGATG

GGGTCTTAGGTGGTGTC 1767

GACACCACCTAAGACCC 1768 ®

GBSS GGTCTTATGTGGTGTAGAGAAGCGCGTGCTTCAATTCCCAGGGAA

Astragalus GAACAGAAGCCAAAGTGAATTCACCTCAGAA membranaeus TTCTGAGGTGAATTCACTTTGGCTTCTGTTCTTCCCTGGGAATTGA | 1770

Ser{1Term AGCACGCGCTTCTCTACACCACATAAGACCCCGTCACCGTTGCCA

TCG-TAG TTCTGTGAGAGAGCAGTAAGGAGCAACAAT

TGTGGTGTAGAGAAGCG

CGCTTCTCTACACCACA 1772 1142101 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration

Waxy starch TGTTGCTCCTTACTGCTCTCTCACAGAATGGCAACGGTGACGGGG | 1773

GBSS TCTTATGTGGTGTCGTGAAGCGCGTGCTTCAATTCCCAGGGAAGA

‘ Astragalus ACAGAAGCCAAAGTGAATTCACCTCAGAAGA membranaeus TCTTCTGAGGTGAATTCACTTTGGCTTCTGTTCTTCCCTGGGAATT | 1774

Arg12Term GAAGCACGCGCTTCACGACACCACATAAGACCCCGTCACCGTTGC

AGA-TGA CATTCTGTGAGAGAGCAGTAAGGAGCAACA

TGGTGTCGIGAAGCGCG

CGCGCTTCACGACACCA 1776

Waxy starch ACTGCTCTCTCACAGAATGGCAACGGTGACGGGGTCTTATGTGGT | 1777 ® GBSS GTCGAGAAGCGCGTGATTCAATTCCCAGGGAAGAACAGAAGCCAA

Astragalus AGTGAATTCACCTCAGAAGATAAATCTCAAT membranaeus ATTGAGATTTATCTTCTGAGGTGAATTCACTTTGGCTTCTGTTCTTC | 1778

Cys15Term CCTGGGAATTGAATCACGCGCTTCTCGACACCACATAAGACCCCG

TGC-TGA TCACCGTTGCCATTCTGTGAGAGAGCAGT

AGCGCGTGATTCAATTC 1779

GAATTGAATCACGCGCT 1780

GBSS CGCGTGCTTCAATTCCTAGGGAAGAACAGAAGCCAAAGTGAATTC

Astragalus ACCTCAGAAGATAAATCTCAATAGCCAAGCAT membranaeus ATGCTTGGCTATTGAGATTTATCTTCTGAGGTGAATTCACTTTGGCT| 1782

GIn19Term TCTGTTCTTCCCTAGGAATTGAAGCACGCGCTTCTCGACACCACAT

CAG-TAG AAGACCCCGTCACCGTTGCCATTCTGTG

TCAATTCCTAGGGAAGA 1783

TCTTCCCTAGGAATTGA

® Waxy starch TGTAGCTTGGTAGATTCCCCTTTTTGTAGACCACACATCACATGGC | 1785

GBSS AAGCATCACAGCTTGACACCACTTTGTGTCAAGAAGCCAAACTTCA

Solanum tuberosum |CTAGACACCAAATCAACCTTGTCACAGAT

Ser7Term ATCTGTGACAAGGTTGATTTGGTGTCTAGTGAAGTTTGGCTTCTTG | 1786

TCA-TGA ACACAAAGTGGTGTCAAGCTGTGATGCTTGCCATGTGATGTGTGG

TCTACAAAAAGGGGAATCTACCAAGCTACA

CACAGCTTGACACCACT 1787

AGTGGTGTCAAGCTGTG 1788 aK Waxy starch TCCCCTTTTTGTAGACCACACATCACATGGCAAGCATCACAGCTTC | 1789

GBSS ACACCACTTTGTGTGAAGAAGCCAAACTTCACTAGACACCAAATCA

. Solanum tuberosum |ACCTTGTCACAGATAGGACTCAGGAACCA

Ser12Term TGGTTCCTGAGTCCTATCTGTGACAAGGTTGATTTGGTGTCTAGTG | 1790

TCA-TGA AAGTTTGGCTTCTTCACACAAAGTGGTGTGAAGCTGTGATGCTTGC

CATGTGATGTGTGGTCTACAAAAAGGGGA

11/12/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

CTTTGTGTGAAGAAGCC 1791 i GGCTTCTTCACACAAAG 1792

Waxy starch CCCTTTTTGTAGACCACACATCACATGGCAAGCATCACAGCTTCAC | 1793

GBSS ACCACTTTGTGTCATGAAGCCAAACTTCACTAGACACCAAATCAAC

Solanum tuberosum |CTTGTCACAGATAGGACTCAGGAACCATA

Arg13Term TATGGTTCCTGAGTCCTATCTGTGACAAGGTTGATTTGGTGTCTAG | 179%

AGA-TGA TGAAGTTTGGCTTCATGACACAAAGTGGTGTGAAGCTGTGATGCTT

GCCATGTGATGTGTGGTCTACAAAAAGGG

TTGTGTCATGAAGCCAA 1795 ® TTGGCTTCATGACACAA 1796

Waxy starch TTGTAGACCACACATCACATGGCAAGCATCACAGCTTCACACCACT | 1797

GBSS TTGTGTCAAGAAGCTAAACTTCACTAGACACCAAATCAACCTTGTC

Solanum tuberosum |ACAGATAGGACTCAGGAACCATACTCTGA

Gin15Term TCAGAGTATGGTTCCTGAGTCCTATCTGTGACAAGGTTGATTTGGT | 1798

CAA-TAA GTCTAGTGAAGTTTAGCTTCTTGACACAAAGTGGTGTGAAGCTGTG

ATGCTTGCCATGTGATGTGTGGTCTACAA

CAAGAAGCTAAACTTCA

TGAAGTTTAGCTTCTTG 1800 starch CCACACATCACATGGCAAGCATCACAGCTTCACACCACTTTIGTGTC | 1801

GBSS AAGAAGCCAAACTTGACTAGACACCAAATCAACCTTGTCACAGATA

Solanum tuberosum |GGACTCAGGAACCATACTCTGACTCACAA

Ser17Term TTGTGAGTCAGAGTATGGTTCCTGAGTCCTATCTGTGACAAGGTTG | 1802

TCA-TGA ATTTGGTGTCTAGTCAAGTTTGGCTTCTTGACACAAAGTGGTGTGA ® AGCTGTGATGCTTGCCATGTGATGTGTGG

CCAAACTTGACTAGACA 1803

TGTCTAGTCAAGTTTGG 1804

GBSS TGGCAACAATAACGTGATCTTCAATGCCGACGAGAACCGCGTGCT

Pisum sativum TCAATTACCAAGGAAGATCAGCAGAGTCTA

Gly6Term TAGACTCTGCTGATCTTCCTTGGTAATTGAAGCACGCGGTTCTCGT | 1806

GGA-TGA CGGCATTGAAGATCACGTTATTGTTGCCATTTTTGTGTCAAAATCT

GTTTCGGCGGTGAGAGAAGAGTGATCGAC

' CAATAACGIGATCTTCA 1807

TGAAGATCACGTTATTG 1808

N20! 01:47 pm 03137.009 — [NY)793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos )

Waxy starch ACTCTTCTCTCACCGCCGAAACAGATTTTGACACAAAAATGGCAAC | 1809

GBSS AATAACGGGATCTTGAATGCCGACGAGAACCGCGTGCTTCAATTA

) Pisum sativum CCAAGGAAGATCAGCAGAGTCTAAACTGAA

Ser8Term TTCAGTTTAGACTCTGCTGATCTTCCTTGGTAATTGAAGCACGCGG | 1810

TCA-TGA TTCTCGTCGGCATTCAAGATCCCGTTATTGTTGCCATTTTTGTGTCA

AAATCTGTTTCGGCGGTGAGAGAAGAGT

GGGATCTTGAATGCCGA 1811

TCGGCATTCAAGATCCC 1812

Waxy starch ACCGCCGAAACAGATTTTGACACAAAAATGGCAACAATAACGGGA | 1813 ® GBSS TCTTCAATGCCGACGIGAACCGCGTGCTTCAATTACCAAGGAAGA

Pisum sativum TCAGCAGAGTCTAAACTGAATTTGCCTCAGA

Arg12Term TCTGAGGCAAATTCAGTTTAGACTCTGCTGATCTTCCTTGGTAATT 1814

AGA-TGA GAAGCACGCGGTTCACGTCGGCATTGAAGATCCCGTTATTGTTGC

CATTTTTGTGTCAAAATCTGTTTCGGCGGT

TGCCGACGTGAACCGCG

CGCGGTTCACGTCGGCA 1816

Waxy starch AGATTTTGACACAAAAATGGCAACAATAACGGGATCTTCAATGCCG | 1817

GBSS ACGAGAACCGCGTGATTCAATTACCAAGGAAGATCAGCAGAGTCT

Pisum sativum AAACTGAATTTGCCTCAGATACACTTCAAT

Cys15Term ATTGAAGTGTATCTGAGGCAAATTCAGTTTAGACTCTGCTGATCTT | 1818

TGC-TGA CCTTGGTAATTGAATCACGCGGTTCTCGTCGGCATTGAAGATCCC

GTTATTGTTGCCATTTTTGTGTCAAAATCT

ACCGCGTGATTCAATTA 1819

TAATTGAATCACGCGGT

® AAAAATGGCAACAATAACGGGATC TTGAATGCCGACGAGAAC | 1821

GBSS CGCGTGCTTCAATTAGCAAGGAAGATCAGCAGAGTCTAAACTGAA

Pisum sativum TTTGCCTCAGATACACTTCAATAACAACCAA

Tyr18Term TTGGTTGTTATTGAAGTGTATCTGAGGCAAATTCAGTTTAGACTCT 1822

TAC-TAG GCTGATCTTCCTTGCTAATTGAAGCACGCGGTTCTCGTCGGCATTG

AAGATCCCGTTATTGTTGCCATTTTTGTG

TTCAATTAGCAAGGAAG 1823

CTTCCTTGCTAATTGAA 1824

GBSS TCGTTTCCAGGAGCTGACACTTGAGCATCCATGCATTAGAGACTAA

A Manihot esculenta GGCTAATAATTTGTCTCACACTGGACCCTG

Ser14Term CAGGGTCCAGTGTGAGACAAATTATTAGCCTTAGTCTCTAATGCAT | 1826

TCA-TGA GGATGCTCAAGTGTCAGCTCCTGGAAACGAAATGTGCAGCTATTA

CAGTTGCCATGGTGCTCTCTCCGGTGTAGA

1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos , Alteration

CAGGAGCTGACACTTGA 1827 } TCAAGTGTCAGCTCCTG 1828

Waxy starch CCGGAGAGAGCACCATGGCAACTGTAATAGCTGCACATTTCGTTT | 1829

GBSS CCAGGAGCTCACACTAGAGCATCCATGCATTAGAGACTAAGGCTA

Manihot esculenta ATAATTTGTCTCACACTGGACCCTGGACCCA

Leu16Term TGGGTCCAGGGTCCAGTGTGAGACAAATTATTAGCCTTAGTCTCTA| 1830

TTG-TAG ATGCATGGATGCTCTAGTGTGAGCTCCTGGAAACGAAATGTGCAG

CTATTACAGTTGCCATGGTGCTCTCTCCGG

CTCACACTAGAGCATCC 1831 ® GGATGCTCTAGTGTGAG 1832

Waxy starch | TGGCAACTGTAATAGCTGCACATTTCGTTTCCAGGAGGTCACACTT | 1833

GBSS GAGCATCCATGCATGAGAGACTAAGGCTAATAATTTGTCTCACACT

Manihot esculenta GGACCCTGGACCCAAACTATCACTCCCAA

Leu21Term TTGGGAGTGATAGTTTGGGTCCAGGGTCCAGTGTGAGACAAATTA | 1834

TTA-TGA TTAGCCTTAGTCTCTCATGCATGGATGCTCAAGTGTGAGCTCCTGG

AAACGAAATGTGCAGCTATTACAGTTGCCA

CCATGCATGAGAGACTA

TAGTCTCTCATGCATGG

GBSS GCATCCATGCATTATAGACTAAGGCTAATAATTTGTCTCACACTGG

Manihot esculenta ACCCTGGACCCAAACTATCACTCCCAATG

Glu22Term CATTGGGAGTGATAGTTTGGGTCCAGGGTCCAGTGTGAGACAAAT | 1838

GAG-TAG TATTAGCCTTAGTCTATAATGCATGGATGCTCAAGTGTGAGCTCCT ® GGAAACGAAATGTGCAGCTATTACAGTTGC

ATGCATTATAGACTAAG 1839

CTTAGTCTATAATGCAT 1840

GBSS ATGCATTAGAGACTTAGGCTAATAATTTGTCTCACACTGGACCCTG

Manihot esculenta GACCCAAACTATCACTCCCAATGGTTTAA

Lys24Term TTAAACCATTGGGAGTGATAGTTTGGGTCCAGGGTCCAGTGTGAG | 1842

AAG-TAG ACAAATTATTAGCCTAAGTCTCTAATGCATGGATGCTCAAGTGTGA

GCTCCTGGAAACGAAATGTGCAGCTATTAC

. TAGAGACTTAGGCTAAT 1843

ATTAGCCTAAGTCTCTA 1844 1112/01 01:47 pm 03137.009 — [NY]793558.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Waxy starch ACAACTCCTCCGTCACCGGTATAAGCATGGCAACGGTATCGATGG | 1845

GBSS CATCGTGCGTGGCGTGAAAAGGCGCGTGGAGTACAGAGACAAAA i Phaseolus vulgaris GTGAAATCTTCGGGTCAGATGAGCCTGAACCG

Ser12Term CGGTTCAGGCTCATCTGACCCGAAGATTTCACTTTTGTCTCTGTAC | 1846

TCA-TGA TCCACGCGCCTTTTCACGCCACGCACGATGCCATCGATACCGTTG

CCATGCTTATACCGGTGACGGAGGAGTTGT

CGTGGCGTGAAAAGGCG 1847

CGCCTTTTCACGCCACG

Waxy starch CACCGGTATAAGCATGGCAACGGTATCGATGGCATCGTGCGTGGC | 1849 ® GBSS GTCAAAAGGCGCGTGAAGTACAGAGACAAAAGTGAAATCTTCGGG

Phaseolus vulgaris ~~ |TCAGATGAGCCTGAACCGTCATGAATTGAAA

Trp16Term TTTCAATTCATGACGGTTCAGGCTCATCTGACCCGAAGATTTCACT

TGG-TGA TTTGTCTCTGTACTTICACGCGCCTTTTGACGCCACGCACGATGCCA

TCGATACCGTTGCCATGCTTATACCGGTG

GGCGCGTGAAGTACAGA | 1851

TCTGTACTICACGCGCC 1852

Waxy starch ATAAGCATGGCAACGGTATCGATGGCATCGTGCGTGGCGTCAAAA | 1853

GBSS GGCGCGTGGAGTACATAGACAAAAGTGAAATCTTCGGGTCAGATG

Phaseolus vulgaris AGCCTGAACCGTCATGAATTGAAATACGATG

Glu19Term CATCGTATTTCAATTCATGACGGTTCAGGCTCATCTGACCCGAAGA

GAG-TAG TTTCACTTTTGTCTATGTACTCCACGCGCCTTTTGACGCCACGCAC

GATGCCATCGATACCGTTGCCATGCTTAT

GGAGTACATAGACAAAA 1855

TTTITGTCTATGTACTCC | 1856 ® ARAAGGCGC | 1857

GBSS GTGGAGTACAGAGACATAAGTGAAATCTTCGGGTCAGATGAGCCT

Phaseolus vulgaris GAACCGTCATGAATTGAAATACGATGGGTTGA

Lys21Term TCAACCCATCGTATTTCAATTCATGACGGTTCAGGCTCATCTGACC | 1858

AAA-TAA CGAAGATTTCACTTATGTCTCTGTACTCCACGCGCCTTTTGACGCC

ACGCACGATGCCATCGATACCGTTGCCAT

CAGAGACATAAGTGAAA 1859

TTTCACTTATGTCTCTG 1860

ARAAGGCGCGTGGAG | 1861

GBSS TACAGAGACAAAAGTGTAATCTTCGGGTCAGATGAGCCTGAACCG

] Phaseolus vulgaris TCATGAATTGAAATACGATGGGTTGAGATCTC

Lys23Term GAGATCTCAACCCATCGTATTTCAATTCATGACGGTTCAGGCTCAT | 1862

AAA-TAA CTGACCCGAAGATTACACTTTTGTCTCTGTACTCCACGCGCCTTTT

GACGCCACGCACGATGCCATCGATACCGT

11/12/01 01:47 pm 03137.009 — [NY)793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

CAAAAGTGTAATCTTCG 1863 . CGAAGATTACACTTTTG 1864

Waxy starch GCGCCTAGCTCGAAAAGGTCGTCATTGAGAGGCTGCACCAATGG 1865

GBSS GTTCCATTCCTAATTAGTGTTCTTATCAAACAAACAGTGTTGGTTCA

Triticum aestivum CTGAAACTGTCGCCTCACATCCAATTCCAG

Tyr7Term CTGGAATTGGATGTGAGGCGACAGTTTCAGTGAACCAACACTGTT | 1866

TAT-TAG TGTTTGATAAGAACACTAATTAGGAATGGAACCCATTGGTGCAGCC

TCTCAATGACGACCTTTTCGAGCTAGGCGC

CCTAATTAGTGTTCTTA 1867 ® TAAGAACACTAATTAGG 1868

Waxy starch CCTAGCTCGAAAAGGTCGTCATTGAGAGGCTGCACCAATGGGTTC | 1869

GBSS CATTCCTAATTATTGATCTTATCAAACAAACAGTGTTGGTTCACTGA

Triticum aestivum AACTGTCGCCTCACATCCAATTCCAGCAA

Cys8Term TTGCTGGAATTGGATGTGAGGCGACAGTTTCAGTGAACCAACACT | 1870

TGT-TGA GTTTGTTTGATAAGATCAATAATTAGGAATGGAACCCATTGGTGCA

GCCTCTCAATGACGACCTTTTCGAGCTAGG

TGATAAGATCAATAATT 1872

Waxy starch TCGAAAAGGTCGTCATTGAGAGGCTGCACCAATGGGTTCCATTCC | 1873

GBSS TAATTATTGTTCTTAGCAAACAAACAGTGTTGGTTCACTGAAACTGT

Triticum aestivum CGCCTCACATCCAATTCCAGCAATCTTGT

Tyr10Term TACAAGATTGCTGGAATTGGATGTGAGGCGACAGTTTCAGTGAACC | 1874

TAT-TAG AACACTGTTTGTTTGCTAAGAACAATAATTAGGAATGGAACCCATT ® GGTGCAGCCTCTCAATGACGACCTTTTCGA

TGTTCTTAGCAAACAAA 1875

TTTGTTTGCTAAGAACA 1876

Waxy starch CGAAAAGGTCGTCATTGAGAGGCTGCACCAATGGGTTCCATTCCT | 1877

GBSS AATTATTGTTCTTATIAAACAAACAGTGTTGGTTCACTGAAACTGTC

Triticum aestivum GCCTCACATCCAATTCCAGCAATCTTGTA

GIn11Term TACAAGAT GCTGGAATTGGATGTGAGGCGACAGTTTCAGTGAAC | 1878 | -

CAA-TAA CAACACTGTTTGTTTAATAAGAACAATAATTAGGAATGGAACCCATT

GGTGCAGCCTCTCAATGACGACCTTTTCG

. GTTCTTATTAAACAAAC 1879

GTTTGTTTAATAAGAAC 1880 : 1112001 01:47 pm 03137.008 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Waxy starch AGGCTGCACCAATGGGTTCCATTCCTAATTATTGTTCTTATCAAACA] 1881

GBSS AACAGTGTTGGTTGACTGAAACTGTCGCCTCACATCCAATTCCAGC

’ Triticum aestivum AATCTTGTAACAATGAAGTTATGTTCCT

Ser17Term AGGAACATAACTTCATTGTTACAAGATTGCTGGAATTGGATGTGAG | 1882

TCA-TGA GCGACAGTTTCAGTCAACCAACACTGTTTGTTTGATAAGAACAATA

ATTAGGAATGGAACCCATTGGTGCAGCCT

TGTTGGTTGACTGAAAC 1883

GTTTCAGTCAACCAACA 1884

Waxy starch CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT | 1885 ® GBSS CCGGCGTGCAGGTTTCTAGGGCGTGAGGCCCCGGAGCCCGGCG

Triticum aestivum GATGCGGCTCTCGGCATGAGGACCGTCGGAGCTA

GIn28Term TAGCTCCGACGGTCCTCATGCCGAGAGCCGCATCCGCCGGGCTC | 1886

CAG-TAG CGGGGCCTCACGCCCTAGAAACCTGCACGCCGGAACCTGTCGGT

GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG

CAGGTTTCTAGGGCGTG 1887

CACGCCCTAGAAACCTG 1888

Waxy starch ~ [GGTTTCCAGGGCGTGAGGCCCCGGAGCCCGGCGGATGCGGCTCT| 1889

GBSS CGGCATGAGGACCGTCTGAGCTAGCGCCGCCCCAACGCAAAGCC

Triticum aestivum GGAAAGCGCACCGCGGGACCCGGCGGTGCCTCT

Gly46Term AGAGGCACCGCCGGGTCCCGCGGTGCGCTTTCCGGCTTTGCGTT | 1890

GGA-TGA GGGGCGGCGCTAGCTCAGACGGTCCTCATGCCGAGAGCCGCATC

CGCCGGGCTCCGGGGCCTCACGCCCTGGAAACC

GGACCGTCIGAGCTAGC 1891

GCTAGCTCAGACGGTCC 1892 ® Waxy starch CGGAGCCCGGCGGATGCGGCTCTCGGCATGAGGACCGTCGGAG | 1893

GBSS CTAGCGCCGCCCCAACGTAAAGCCGGAAAGCGCACCGCGGGACC

Triticum aestivum CGGCGGTGCCTCTCCATGGTGGTGCGCGCCACCG

GIn53Term CGGTGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCG | 1894

CAA-TAA GTGCGCTTTCCGGCTTTACGTTGGGGCGGCGCTAGCTCCGACGG

TCCTCATGCCGAGAGCCGCATCCGCCGGGCTCCG

CCCCAACGTAAAGCCGG 1895

CCGGCTTTACGTTGGGG 1896

GBSS CCCAACGCAAAGCCGGTAAGCGCACCGCGGGACCCGGCGGTGC

} Triticum aestivum CTCTCCATGGTGGTGCGCGCCACCGGCAGCGGCG

Lys56Term CGCCGCTGCCGGTGGCGCGCACCACCATGGAGAGGCACCGCCG | 1898

AAA-TAA GGTCCCGCGGTGCGCTTACCGGCTTTGCGTTGGGGCGGCGCTAG

CTCCGACGGTCCTCATGCCGAGAGCCGCATCCGC

1112/01 01:47 pm 03137.009 — |NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration ——

AAAGCCGGTAAGCGCAC

} GTGCGCTTACCGGCTTT 1900 1]

Waxy starch CTCTCCATGGTGGTGCGCGCCACCGGCAGCGGCGGCATGAACCT | 1901

GBSS CGTGTTCGTCGGCGCCTAGATGGCGCCCTGGAGCAAGACCGGCG

Triticum aestivum GCCTCGGCGACGTCCTCGGGGGCCTCCCCCCAG

Glu85Term CTGGGGGGAGGCCCCCGAGGACGTCGCCGAGGCCGCCGGTCTT | 1902

GAG-TAG GCTCCAGGGCGCCATCTAGGCGCCGACGAACACGAGGTTCATGC

CGCCGCTGCCGGTGGCGCGCACCACCATGGAGAG

TCGGCGCCTAGATGGCG 1903 [ CGCCATCTAGGCGCCGA 1904

Waxy starch GTCGTCTCTCGCTGCAGGTAGCCACACCCTGCGCGCGCGATGGC | 1905

GBSS GGCTCTGGTCACGTCGIAGCTCGCCACCTCCGGCACCGTCCTCG

Triticum aestivum GCATCACCGACAGGTTCCGGCGTGCAGGTTTTC

GIn8Term GAAAACCTGCACGCCGGAACCTGTCGGTGATGCCGAGGACGGTG

CAG-TAG CCGGAGGTGGCGAGCTACGACGTGACCAGAGCCGCCATCGCGC

GCGCAGGGTGTGGCTACCTGCAGCGAGAGACGAC

TCACGTCGTAGCTCGCC

GGCGAGCTACGACGTGA 1908

Waxy starch CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT | 1909

GBSS CCGGCGTGCAGGTTTTTAGGGTGTGAGGCCCCGGAGCCCGGCAG

Triticum aestivum ATGCGCCGCTCGGCATGAGGACTACCGGAGCGA

GIn28Term TCGCTCCGGTAGTCCTCATGCCGAGCGGCGCATCTGCCGGGCTC | 1910

CAG-TAG CGGGGCCTCACACCCTAAAAACCTGCACGCCGGAACCTGTCGGT ® GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG

CACACCCTAAAAACCTG 1912

Waxy starch CCCCGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGG] 1913

GBSS AGCGAGCGCCGCCCCGTAGCAACAAAGCCGGAAAGCGCACCGCG

Triticum aestivum GGACCCGGCGGTGCCTCTCCATGGTGGTGCGCEG

Lys52Term CGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGRTRCGE | 1914

AAG-TAG TTTCCGGCTTTGTTGCTACGGGGCGGCGCTCGCTCCGRTAGTCCT

CATGCCGAGCGGCGCATCTGCCGGGCTCCGGGG

: CCGCCCCGTAGCAACAA 1915

TTGTIGCTACO068066 | 1st

YY] 11712001 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene, ]

Plant & Targeted Altering Oligos __ Alteration} 00}

Waxy starch CGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAG 1917

GBSS CGAGCGCCGCCCCGAAGTAACAAAGCCGGAAAGCGCACCGCGEG

’ Triticum aestivum GACCCGGCGGTGCCTCTCCATGGTGGTGCGCGCCA

GIn53Term TGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGRTG

CAA-TAA CGCTTTCCGGCTTTGTTACTTCGGGGCGGCGCTCGCTCCGGTAGT

CCTCATGCCGAGCGGCGCATCTGCCGGGCTCCG

CCCCGAAGTAACAAAGC 1919

GCTTTGTTACTTCGGGG

Waxy starch N AGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAGCGAG | 1921 ® GBSS CGCCGCCCCGAAGCAATAAAGCCGGAAAGCGCACCGCGGGACCC

Triticum aestivum GGCGGTGCCTCTCCATGGTGGTGCGCGCCACGG

GIn54Term CCGTGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCa 1922

CAA-TAA GTGCGCTTTCCGGCTTTATTGCTTCGGGGCGGCGCTCGCTCCGRT

AGTCCTCATGCCGAGCGGCGCATCTGCCGGGCT

CGAAGCAATAAAGCCGG 1923

CCGGCTTTATTGCTTCG 1924

Waxy starch CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT 1925

GBSS CCGGCGTGCAGGTTTCTAGGGCGTGAGGCCCCGGAACCCGGCG

Triticum durum GATGCGGCCCTCGTCATGAGGACTATCGGAGCGA

GIn28Term TCGCTCCGATAGTCCTCATGACGAGGGCCGCATCCGCCGGGTTC 1926

CAG-TAG CGGGGCCTCACGCCCTAGAAACCTGCACGCCGGAACCTGTCGGT

GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG

CAGGTTTCTAGGGCGTG 1927

CACGCCCTAGAAACCTG 1928 ®

GBSS AGCGAGCGCCGCCCCGTAGCAAAGCCGGAAAGCGCACCGCGGG

Triticum durum AGCCGGCGGTGCCTCTCCATGGTGGTGCGCGCCA

Lys52Term TGGCGCGCACCACCATGGAGAGGCACCGCCGGCTCCCGCGRTG 1930

AAG-TAG CGCTTTCCGGCTTTGCTACGGGGCGGCGCTCGCTCCGATAGTCCT

CATGACGAGGGCCGCATCCGCCGGGTTCCGGGE

CCGCCCCGTAGCAAAGC 1931

GBSS GAGCGCCGCCCCGAAGTAAAGCCGGAAAGCGCACCGCGGGAGE

} Triticum durum CGGCGGTGCCTCTCCATGGTGGTGCGCGCCACGG

GIn53Term CCGTGGCGCGCACCACCATGGAGAGGCACCGCCGGCTCCCGLE 1934

CAA-TAA GTGCGCTTTCCGGCTTTACTTCGGGGCGGCGCTCGCTCCGATAGT

CCTCATGACGAGGGCCGCATCCGCCGGGTTCCG

1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797 —_—

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

CCCCGAAGTAAAGCCGG 1935

CCGGCTTTACTTCGGGG 1936

Waxy starch GCGGATGCGGCCCTCGTCATGAGGACTATCGGAGCGAGCGCCGC | 1937

GBSS CCCGAAGCAAAGCCGGTAAGCGCACCGCGGGAGCCGGCGGRTGC

Triticum durum CTCTCCATGGTGGTGCGCGCCACGGGCAGCGGCG

Lys56Term CGCCGCTGCCCGTGGCGCGCACCACCATGGAGAGGCACCGCCG | 1938

AAA-TAA GCTCCCGCGGTGCGCTTACCGGCTTTGCTTCGGGGCGGCGCTCG

CTCCGATAGTCCTCATGACGAGGGCCGCATCCGC

{AAAGCCGGTAAGCGCAC 1939 ® GTGCGCTTACCGGCTTT 1940

GBSS GCGGGAGCCGGCGGTGACTCTCCATGGTGGTGCGCGCCACGGG

Triticum durum CAGCGGCGGCATGAACCTCGTGTTCGTCGGCGCC

Cys64Term GGCGCCGACGAACACGAGGTTCATGCCGCCGCTGCCCGTGGCGC | 1942

TGC-TGA GCACCACCATGGAGAGICACCGCCGGCTCCCGCGGTGCGLCTTTC

CGGCTTTGCTTCGGGGCGGCGCTCGCTCCGATA

CGGCGGTGACTCTCCAT 1943

ATGGAGAGICACCGCCG 1944

GBSS CCGGCGTGCAGGTTTTTAGGGTGTGAGGCCCCGGAGCCCGGCAG "

Triticum turgidum ATGCGCCGCTCGGCATGAGGACTACCGGAGCGA

Gin28Term TCGCTCCGGTAGTCCTCATGCCGAGCGGCGCATCTGCCGGGCTC | 1946

CAG-TAG CGGGGCCTCACACCCTAAAAACCTGCACGCCGGAACCTGTCGGT

GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG

® CAGGTTTTTAGGGTGTG

CACACCCTAAAAACCTG 1948

Waxy starch | CCOCGGAGCCCGGCAGATGCGLCGCTCOGCATGAGGACTACCGE | 1048

GBSS AGCGAGCGCCGCCCCGTAGCAACAAAGCCGGAAAGCGCACCGCEG

Triticum turgidum GGACCCGGCGGTGCCTCTCCATGGTGGTGCGCG

Lys52Term CGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGGTGCGC | 1950

AAG-TAG TTTCCGGCTTTGTTGCTACGGGGCGGCGCTCGCTCCGGTAGTCCT

CATGCCGAGCGGCGCATCTGCCGGGCTCCGGGG

: CCGCCCCGTAGCAACAA 1951 [TTGTTGOTACGo66C66 | foi 1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos :

Waxy starch CGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAG | 1953

GBSS CGAGCGCCGCCCCGAAGTAACAAAGCCGGAAAGCGCACCGCGE

Triticum turgidum GACCCGGCGGTGCCTCTCCATGGTGGTGCGCGCCA

GinS3Term TGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGGTG | 1954

CAA-TAA CGCTTTCCGGCTTTGTTACTTCGGGGCGGCGCTCGCTCCGGTAGT

CCTCATGCCGAGCGGCGCATCTGCCGGGCTCCG

CCCCGAAGTAACAAAGC

GCTTTGTTACTTCGGGG 1956

Waxy starch AGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAGCGAG| 1957 ® GBSS CGCCGCCCCGAAGCAATAAAGCCGGAAAGCGCACCGCGGGACCC

Triticum turgidum GGCGGTGCCTCTCCATGGTGGTGCGCGCCACGG

GIn54Term CCGTGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCG

CAA-TAA GTGCGCTTTCCGGCTTTATTGCTTCGGGGCGGCGCTCGCTCCGGT

AGTCCTCATGCCGAGCGGCGCATCTGCCGGGCT

CGAAGCAATAAAGCCGG 1959

CCGGCTTTATTGCTTCG 1960

Waxy starch GATGCGCCGCTCGGCATGAGGACTACCGGAGCGAGCGCCGCCCC| 1961

GBSS GAAGCAACAAAGCCGGTAAGCGCACCGCGGGACCCGGCGGTGC

Triticum turgidum CTCTCCATGGTGGTGCGCGCCACGGGCAGCGCCG

Lys57Term CGGCGCTGCCCGTGGCGCGCACCACCATGGAGAGGCACCGCCG | 1962

AAA-TAA GGTCCCGCGGTGCGCTTACCGGCTTTGTTGCTTCGGGGCGGCGC

TCGCTCCGGTAGTCCTCATGCCGAGCGGCGCATC

AAAGCCGGTAAGCGCAC 1963

GTGCGCTTACCGGCTTT

® Waxy starch CAGCTCGCCACCTCCGCCACCGTCCTCGGCATCACCGACAGGTTC| 1965

GBSS CGCCATGCAGGTTTCTAGGGCGTGAGGCCCCGGAGCCCGGCAGA

Aegilops spelfoides |TGCGCCGCTCGGCATGAGGACTGTCGGAGCGA

GIn28Term TCGCTCCGACAGTCCTCATGCCGAGCGGCGCATCTGCCGGGCTC | 1966

CAG-TAG CGGGGCCTCACGCCCTAGAAACCTGCATGGCGGAACCTGTCGGT

GATGCCGAGGACGGTGGCGGAGGTGGCGAGCTG

CAGGTTTCTAGGGCGTG 1967

CACGCCCTAGAAACCTG 1968 ’ Waxy starch GGTTTCCAGGGCGTGAGGCCCCGGAGCCCGGCAGATGCGCCGCT| 1969

GBSS CGGCATGAGGACTGTCTGAGCGAGCGCCGCCCCGAAGCAACAAA

. Aegilops speltoides ~~ |GCCGGAAAGCGCACCGCGGGACCCGGCGGTGCC

Glyd6Term GGCACCGCCGGGTCCCGCGGTGCGCTTTCCGGCTTTGTTGCTTC | 1970

GGA-TGA GGGGCGGCGCTCGCTCAGACAGTCCTCATGCCGAGCGGCGCATC

TGCCGGGCTCCGGGGCCTCACGCCCTGGAAACC

1112/01 01:47 pm 03137.009 — {NY}793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ___ Alteration

GGACTGTCTGAGCGAGC 1971

GCTCGCTCAGACAGTCC 1972

Waxy starch CCCCGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTGTCGG| 1973

GBSS AGCGAGCGCCGCCCCGTAGCAACAAAGCCGGAAAGCGCACCGCG

Aegilops spelfoides GGACCCGGCGGTGCCTCTCGATGGTGGTGCGCG

Lys52Term CGCGCACCACCATCGAGAGGCACCGCCGGGTCCCGCGGTGCGCT| 1974

AAG-TAG TTCCGGCTTTGTTGCTACGGGGCGGCGCTCGCTCCGACAGTCCTC

ATGCCGAGCGGCGCATCTGCCGGGCTCCGGGG

CCGCCCCGTAGCAACAA 1975 ® TTGTTGCTACGGGGCGG 1976

Waxy starch CGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTGTCGGAG | 1977

GBSS CGAGCGCCGCCCCGAAGTAACAAAGCCGGAAAGCGCACCGCGG

Aegilops spelfoides |GACCCGGCGGTGCCTCTCGATGGTGGTGCGCGCCA

GIn53Term TGGCGCGCACCACCATCGAGAGGCACCGCCGGGTCCCGCGRTG 1978

CAA-TAA CGCTTTCCGGCTTTGTTACTTCGGGGCGGCGCTCGCTCCGACAGT

CCTCATGCCGAGCGGCGCATCTGCCGGGCTCCG

CCCCGAAGTAACAAAGC 1979

GCTTTGTTACTTCGGGG 1980

Waxy starch AGCCCGGCAGATGCGCCGCTCGGCATGAGGACTGTCGGAGCGAG | 1981

GBSS CGCCGCCCCGAAGCAATAAAGCCGGAAAGCGCACCGCGGGACCC

Aegilops speltoides GGCGGTGCCTCTCGATGGTGGTGCGCGCCACCG

GIn54Term CGGTGGCGCGCACCACCATCGAGAGGCACCGCCGGGTCCCGLE 1982 156 CAA-TAA GTGCGCTTTCCGGCTTTATTGCTTCGGGGCGGCGCTCGCTCCGAC ® AGTCCTCATGCCGAGCGGCGCATCTGCCGGGCT

CGAAGCAATAAAGCCGG 1983

CCGGCTTTATTGCTTCG 1984

Waxy slarch | AGTGCAGAGATCTTCCACAGCAACAGCTAGACAACCACCATGTCE | 1988

GBSS GCTCTCACCACGTCCTAGCTCGCCACCTCGGCCACCGGCTTICGG

Oryza glaberrima CATCGCTGACAGGTCGGCGCCGTCGTCGCTGC

GIn8Term GCAGCGACGACGGCGCCGACCTGTCAGCGATGCCGAAGCCGGT 1986

CAG-TAG GGCCGAGGTGGCGAGCTAGGACGTGGTGAGAGCCGACATGRTG

GTTGTCTAGCTGTTGCTGTGGAAGATCTCTGCACT

: CCACGTCCTAGCTCGCC 1987

GGCGAGCTAGGACGTGG 1988 1112/01 01:47 pm 03137.009 — [NY)793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Waxy starch TCCACAGCAACAGCTAGACAACCACCATGTCGGCTCTCACCACGT | 1989

GBSS CCCAGCTCGCCACCTAGGCCACCGGCTTCGGCATCGCTGACAGG

Oryza glaberrima TCGGCGCCGTCGTCGCTGCTCCGCCACGGGTT

Ser12Term AACCCGTGGCGGAGCAGCGACGACGGCGCCGACCTGTCAGCGAT | 1990

TCG-TAG GCCGAAGCCGGTGGCCTAGGTGGCGAGCTGGGACGTGGTGAGA

GCCGACATGGTGGTTGTCTAGCTGTTGCTGTGGA

CGCCACCTAGGCCACCG 1991

CGGTGGCCTAGGTGGCG 1992

Waxy starch CGGCTCTCACCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTC | 1993

PS GBSS GGCATCGCTGACAGGTAGGCGCCGTCGTCGCTGCTCCGCCACGG

Oryza glaberrima GTTCCAGGGCCTCAAGCCCCGCAGCCCCGCCGG

Ser22Term CCGGCGGGGCTGCEGGGGCTTGAGGCCCTGGAACCCGTGGCGGA | 1994

TCG-TAG GCAGCGACGACGGCGCCTACCTGTCAGCGATGCCGAAGCCGGTG

GCCGAGGTGGCGAGCTGGGACGTGGTGAGAGCCG

TGACAGGTAGGCGCCGT 1995

ACGGCGCCTACCTGTCA | 199

Waxystach ~~ JCCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCT | 1997

GBSS GACAGGTCGGCGCCGTAGTCGCTGCTCCGCCACGGGTTCCAGGG

Oryza glaberrima CCTCAAGCCCCGCAGCCCCGCCGGCGGCGACGE

Ser25Term GCGTCGCCGCCGGCGGGGCTGCGGGGCTTGAGGCCCTGGAACC | 1998

TCG-TAG CGTGGCGGAGCAGCGACTACGGCGCCGACCTGTCAGCGATGCCG

AAGCCGGTGGCCGAGGTGGCGAGCTGGGACGTGG

GGCGCCGTAGTCGCTGC 1999

GCAGCGACTACGGCGCC 2000 ® Waxy starch CGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCTGAC | 2001

GBSS AGGTCGGCGCCGTCGTAGCTGCTCCGCCACGGGTTCCAGGGCCT

Oryza glaberrima CAAGCCCCGCAGCCCCGCCGGCGGCGACGCGAC

Ser26Term GTCGCGTCGCCGCCGGCGGGGCTGCGGGGCTTGAGGCCCTGGA | 2002

TCG-TAG ACCCGTGGCGGAGCAGCTACGACGGCGCCGACCTGTCAGCGATG

CCGAAGCCGGTGGCCGAGGTGGCGAGCTGGGACG

GCCGTCGTAGCTGCTCC

GGAGCAGCTACGACGGC 2004 : Waxy starch TCCACAGCAAGAGCTAAACAGCCGACCGTGTGCACCACCATGTCG | 2005

GBSS GCTCTCACCACGTCCTAGCTCGCCACCTCGGCCACCGGCTTCGG

Oryza sativa CATCGCCGACAGGTCGGCGCCGTCGTCGCTGC

GIn8Term GCAGCGACGACGGCGCCGACCTGTCGGCGATGCCGAAGCCGGT | 2006

CAG-TAG GGCCGAGGTGGCGAGCTAGGACGTGGTGAGAGCCGACATGGTG

GTGCACACGGTCGGCTGTTTAGCTCTTGCTGTGGA

1112/01 01:47 pm 03137.009 — {NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

CCACGTCCTAGCTCGCC 2007 } GGCGAGCTAGGACGTGG 2008

Waxy starch CTAAACAGCCGACCGTGTGCACCACCATGTCGGCTCTCACCACGT | 2009

GBSS CCCAGCTCGCCACCTAGGCCACCGGCTTCGGCATCGCCGACAGG

Oryza sativa TCGGCGCCGTCGTCGCTGCTTCGCCACGGGTT

Ser12Term AACCCGTGGCGAAGCAGCGACGACGGCGCCGACCTGTCGGCGAT| 2010

TCG-TAG GCCGAAGCCGGTGGCCIAGGTGGCGAGCTGGGACGTGGTGAGA

GCCGACATGGTGGTGCACACGGTCGGCTGTTTAG

CGCCACCTAGGCCACCG 2011

C CGGTGGCCTAGGTGGCG 2012

Waxy starch CGGCTCTCACCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTC [| 2013

GBSS GGCATCGCCGACAGGTAGGCGCCGTCGTCGCTGCTTCGCCACGG

Oryza sativa GTTCCAGGGCCTCAAGCCCCGTAGCCCAGCCGG

Ser22Term CCGGCTGGGCTACGGGGCTTGAGGCCCTGGAACCCGTGGCGAA 2014

TCG-TAG GCAGCGACGACGGCGCCTACCTGTCGGCGATGCCGAAGCCGGTG

GCCGAGGTGGCGAGCTGGGACGTGGTGAGAGCCG

CGACAGGTAGGCGCCGT

ACGBCGCCTACCTGTCG [206

Waxy starch CCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCC | 2017

GBSS GACAGGTCGGCGCCGTAGTCGCTGCTTCGCCACGGGTTCCAGGS

Oryza sativa CCTCAAGCCCCGTAGCCCAGCCGGCGGGGACGC

Ser25Term GCGTCCCCGCCGGCTGGGCTACGGGGCTTGAGGCCCTGGAACCC] 2018

TCG-TAG GTGGCGAAGCAGCGACTACGGCGCCGACCTGTCGGCGATGCCGA

AGCCGGTGGCCGAGGTGGCGAGCTGGGACGTGG

® GocccceTAGTCGCTee | a

GCAGCGACTACGGCGCC 2020

Waxy starch CGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCCGAC | 2021

GBSS AGGTCGGCGCCGTCGTAGCTGCTTCGCCACGGGTTCCAGGGCCT

Oryza sativa CAAGCCCCGTAGCCCAGCCGGCGGGGACGCATC

Ser26Term GATGCGTCCCCGCCGGCTGGGCTACGGGGCTTGAGGCCCTGGAA | 2022

TCG-TAG CCCGTGGCGAAGCAGCTACGACGGCGCCGACCTGTCGGCGATGE

CGAAGCCGGTGGCCGAGGTGGCGAGCTGGGACS

GAAGCAGCTACGACGGC 2024 fret] 1112/01 01:47 pm 03137.009 — [NY]793558.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Waxy starch GTCTCTCACTGCAGGTAGCCACACCCTGTGCGCGGCGCCATGGC | 2025

GBSS GGCTCTGGCCACGTCCTAGCTCGCCACCTCCGGCACCGTCCTCG

Hordeum vulgare GCGTCACCGACAGATTCCGGCGTCCAGGTTTTC

GIn8Term GAAAACCTGGACGCCGGAATCTGTCGGTGACGCCGAGGACGGTG | 2026

CAG-TAG CCGGAGGTGGCGAGCTAGGACGTGGCCAGAGCCGCCATGGCGC

CGCGCACAGGGTGTGGCTACCTGCAGTGAGAGAC

CCACGTCCTAGCTCGCC 2027

GGCGAGCTAGGACGTGG 2028

Waxy starch ATGGCGGCTCTGGCCACGTCCCAGCTCGCCACCTCCGGCACCGT | 2029 ® GBSS CCTCGGCGTCACCGACTGATTCCGGCGTCCAGGTTTTCAGGGCCT

Hordeum vulgare CAGGCCCCGGAACCCGGCGGATGCGGCGCTTG

Arg21Term CAAGCGCCGCATCCGCCGGGTTCCGGGGCCTGAGGCCCTGAAAA | 2030

AGA-TGA CCTGGACGCCGGAATCAGTCGGTGACGCCGAGGACGGTGCCGG

AGGTGGCGAGCTGGGACGTGGCCAGAGCCGCCAT

TCACCGACTGATTCCGG 2031

CCGGAATCAGTCGGTGA 2032

GBSS CCGGCGTCCAGGTTTTTAGGGCCTCAGGCCCCGGAACCCGGCGEG

Hordeum vulgare ATGCGGCGCTTGGTATGAGGACTATCGGAGCAA

GIn28Term TTGCTCCGATAGTCCTCATACCAAGCGCCGCATCCGCCGGGTTCC | 2034

CAG-TAG GGGGCCTGAGGCCCTAAAAACCTGGACGCCGGAATCTGTCGGTG

ACGCCGAGGACGGTGCCGGAGGTGGCGAGCTG

CAGGTTTTTAGGGCCTC 2035

GAGGCCCTAAAAACCTG 2036 ®

GBSS TGGTATGAGGACTATCTGAGCAAGCGCCGCCCCGAAGCAAAGCC

Hordeum vulgare GGAAAGCGCACCGCGGGAGCCGGCGGTGCCTCT

Gly46Term AGAGGCACCGCCGGCTCCCGCGGTGCGCTTTCCGGCTTTGCTTC | 2038

GGA-TGA GGGGCGGCGCTTGCTCAGATAGTCCTCATACCAAGCGCCGCATC

CGCCGGGTTCCGGGGCCTGAGGCCCTGAAAACC

GGACTATCTIGAGCAAGC 2039

GCTTGCTCAGATAGTCC 2040

GBSS AGCAAGCGCCGCCCCGTAGCAAAGCCGGAAAGCGCACCGCGGG

Hordeum vulgare AGCCGGCGGTGCCTCTCCGTGGTGGTGAGCGCCA

Lys52Term TGGCGCTCACCACCACGGAGAGGCACCGCCGGCTCCCGCGGTGC| 2042

AAG-TAG GCTTTCCGGCTTTGCTACGGGGCGGCGCTTGCTCCGATAGTCCTC

ATACCAAGCGCCGCATCCGCCGGGTTCCGGGG

1142/01 01:47 pm } 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration

CCGCCCCGTAGCAAAGC 2043 : GCTTTGCTACGGGGCGG 2044

Waxy starch ACGTCTTTTCTCTCTCTCCTACGCAGTGGATTAATCGGCATGGCGG | 2045

GBSS CTCTGGCCACGTCGTAGCTCGTCGCAACGCGCGCCGGCCTGGGE

Zea mays GTCCCGGACGCGTCCACGTTCCGCCGCGGLE

GIn8Term CGCCGCGGCGGAACGTGGACGCGTCCGGGACGCCCAGGCCGGC | 2046

CAG-TAG GCGCGTTGCGACGAGCTACGACGTGGCCAGAGCCGCCATGCCGA

TTAATCCACTGCGTAGGAGAGAGAGAAAAGACGT

CCACGTCGTAGCTCGTC 2047 @ GACGAGCTACGACGTGG starch GTCGCAACGCGCGCCGGCCTGGGCGTCCCGGACGCGTCCACGTT| 2049

GBSS CCGCCGCGGCGCCGCGTAGGGCCTGAGGGGGGCCCGGGCGTCG

Zea mays GCGGCGGCGGACACGCTCAGCATGCGGACCAGCG

GIn30Term CGCTGGTCCGCATGCTGAGCGTGTCCGCCGCCGCCGACGCCCGG| 2050

CAG-TAG GCCCCCCTCAGGCCCTACGCGGCGCCGCGGCGGAACGTGGACG

CGTCCGGGACGCCCAGGCCGGCGCGCGTTGCGAC

GCGCCGCGTAGGGCCTG

CAGGCCCTACGCGGCGC

Waxy starch TCCCGGACGCGTCCACGTTCCGCCGCGGCGCCGCGCAGGGCCT | 2053

GBSS GAGGGGGGCCCGGGCGTAGGCGGCGGCGGACACGCTCAGCATG

Zea mays CGGACCAGCGCGCGCGCGGCGCCCAGGCACCAGCA

Ser38Term TGCTGGTGCCTGGGCGCCGCGCGCGCGCTGGTCCGCATGCTGAG | 2054

TCG-TAG CGTGTCCGCCGCCGCCIACGCCCGGGCCCCCCTCAGGCCCTGAG

PY CGGCGCCGCGGCGGAACGTGGACGCGTCCGGGA

CCGGGCGTAGGCGGCGG 2055

CCGCCGCCTACGCCCGG 2056

GBSS GCGCGGCGCCCAGGCACTAGCAGCAGGCGCGCCGCGGGGGCAG

Zea mays GTTCCCGTCGCTCGTCGTGTGCGCCAGCGCCGGCA

Ser57Term TGCCGGCGCTGGCGCACACGACGAGCGACGGGAACCTGCCCCC | 2058

CAG-TAG GCGGCGCGCCTGCTGCTAGTGCCTGGGCGCCGCGCGCGCGCTG

GTCCGCATGCTGAGCGTGTCCGCCGCCGCCGACGE

CCAGGCACTAGCAGCAG 2059

CTGCTGCTAGTGCCTGG 2060 1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797 pee

Phenotype, Gene,

Plant & Targeted Altering Oligos } Alteration

Waxy starch TCGGCGGCGGCGGACACGCTCAGCATGCGGACCAGCGCGCGCG | 2061

GBSS CGGCGCCCAGGCACCAGTAGCAGGCGCGCCGCGGGGGCAGGTT

Zea mays CCCGTCGCTCGTCGTGTGCGCCAGCGCCGGCATGA

GIn58Term TCATGCCGGCGCTGGCGCACACGACGAGCGACGGGAACCTGCCC | 2062

CAG-TAG CCGCGGCGCGCCTGCTACTGGTGCCTGGGCGCCGCGCGCGCGL

TGGTCCGCATGCTGAGCGTGTCCGCCGCCGCCGA

GGCACCAGTAGCAGGCG 2063

CGCCTGCTACTGGTGCC 2064 11/1201 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Example 11

Altering fatty acid content of plants ] Improved means to manipulate fatty acid compositions, from biosynthetic or natural plant sources, are needed. For example, ails containing reduced saturated fatty acids are desired for dietary reasons and oils containing increased saturated fatty acids are also needed as alternatives to current sources of highly saturated oil products, such as tropical oils or chemically hydrogenated oils. It would therefore be advantageous to influence directly the production and composition of fatty acids in crop plants.

Higher plants synthesize fafty acids, primarily palmitic, stearic and oleic acids, in the ® plastids (i.e., chloroplasts, proplastids, or other related organelles) as part of the Fatty Acid Synthase (FAS) complex. Fatty acid synthesis is the result of the three enzymatic activities: acyl-ACP elongase, acyl-ACP desaturase and acyl-ACP thioesterases specific for each of palmitoyl-, stearoyl- and oleoyl-ACP.

A variety of enzymes have been identified that influence the relative levels of saturated vs, unsaturated fatty acids in plants. For example, the enzymes stearoyl-acyl carrier protein (stearoyl-ACP) desaturase, oleoyl desaturase and linoleate desaturase produce unsaturated fatty acids from saturated precursors. Similarly, relative enzymatic activities of the various acyl-ACP thioesterases influences the relative acyl-chain composition of the resultant fatty acids. Consequently a reduction or an increase of the activity of these enzymes can alter the properties of oils produced in a plant. In fact, specific targeting of particular enzymatic activities can results in altered levels of particular fatty acids.

The attached tables disclose exemplary oligonucleotides base sequences which can be ® used to generate site-specific mutations in plant genes encoding proteins involved in fatty acid biosynthesis.

Table 22

Oligonucleotides to produce plants with reduced palmitate

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Reduced palmitate TTTGGTGGCAGTGTCTTTGAACGCTTCATCTCCTCGTCATGGTGGC | 2065

Acyl-ACP-thioesterase {CACCTCTGCTACGTAGTCATTCTTTCCTGTACCATCTTCTTCACTTG : Arabidopsis thaliana |ATCCTAATGGAAAAGGCAATAAGATTGG

Ser8Term CCAATCTTATTGCCTTTTCCATTAGGATCAAGTGAAGAAGATGGTA | 2066

TCG-TAG CAGGAAAGAATGACTACGTAGCAGAGGTGGCCACCATGACGAGG

AGATGAAGCGTTCAAAGACACTGCCACCAAA

1112/01 01:47 pm 03137.009 — [NY]793569.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

TGCTACGTAGTCATTCT 2067 } AGAATGACTACGTAGCA 2068

Reduced palmitate GGTGGCAGTGTCTTTGAACGCTTCATCTCCTCGTCATGGTGGCCA | 2069

Acyl-ACP-thioesterase |CCTCTGCTACGTCGTGATTCTTTCCTGTACCATCTTCTTCACTTGAT

Arabidopsis thaliana | CCTAATGGAAAAGGCAATAAGATTGGGTC

Ser9Term GACCCAATCTTATTGCCTTTTCCATTAGGATCAAGTGAAGAAGATG | 2070

TCA-TGA GTACAGGAAAGAATCACGACGTAGCAGAGGTGGCCACCATGACG

AGGAGATGAAGCGTTCAAAGACACTGCCACC

TACGTCGTGATTCTTTC 2071

CY GAAAGAATCACGACGTA

Reduced palmitate ATCTCCTCGTCATGGTGGCCACCTCTGCTACGTCGTCATTCTTTCC | 2073

Acyl-ACP-thioesterase | TGTACCATCTTCTTGACTTGATCCTAATGGAAAAGGCAATAAGATT

Arabidopsis thaliana |GGGTCTACGAATCTTGCTGGACTCAATTC

Ser17Term GAATTGAGTCCAGCAAGATTCGTAGACCCAATCTTATTGCCTTTTC | 2074

TCA-TGA CATTAGGATCAAGTCAAGAAGATGGTACAGGAAAGAATGACGACG

TAGCAGAGGTGGCCACCATGACGAGGAGAT

ATCTTCTTGACTTGATC

GATCAAGTCAAGAAGAT 2076

Reduced palmitate GTGGCCACCTCTGCTACGTCGTCATTCTTTCCTGTACCATCTTCTT | 2077

Acyl-ACP-thioesterase |CACTTGATCCTAATTGAAAAGGCAATAAGATTGGGTCTACGAATCT

Arabidopsis thaliana |TGCTGGACTCAATTCTGCACCTAACTCTG

Gly22Term CAGAGTTAGGTGCAGAATTGAGTCCAGCAAGATTCGTAGACCCAA | 2078

GGA-TGA TCTTATTGCCTTTTCAATTAGGATCAAGTGAAGAAGATGGTACAGG

AAAGAATGACGACGTAGCAGAGGTGGCCAC

® ATCCTAATIGAAAAGGC 2079

GCCTTTTCAATTAGGAT 2080

Reduced palmitate AZ ) A AA ACAZ 08

Acyl-ACP-thioesterase |TACTGCCGCCACGTGATCATTCTTTCCGTTGACTTCCCCTTCTGGG

Garcinia mangostana |GATGCCAAATCGGGCAATCCCGGAAAAGG : Ser8Term CCTTTTCCGGGATTGCCCGATTTGGCATCCCCAGAAGGGGAAGTC | 2082

TCA-TGA AACGGAAAGAATGATCACGTGGCGGCAGTAGCAACCATTGTGGCC

ACAATTAACCAATCAGATCACAAATTCAAGC

CGCCACGTGATCATTCT

AGAATGATCACGTGGCG 2084 11/12/01 01:47 pm 03137.009 — [NY]793589.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ) Alteration

Acyl-ACP-thioesterase |TGCCGCCACGTCATGATTCTTTCCGTTGACTTCCCCTTCTGGGGAT

Garcinia mangostana |GCCAAATCGGGCAATCCCGGAAAAGGGTC

Ser9Term GACCCTTTTCCGGGATTGCCCGATTTGGCATCCCCAGAAGGGGAA | 2086

TCA-TGA GTCAACGGAAAGAATCATGACGTGGCGGCAGTAGCAACCATTGTG

GCCACAATTAACCAATCAGATCACAAATTCA

CACGTCATGATTCTTTC 2087 .

GAAAGAATCATGACGTG 2088

Reduced palmitate CTGATTGGTTAATTGTGGCCACAATGGTTGCTACTGCCGCCACGT | 2089

Acyl-ACP-thioesterase |CATCATTCTTTCCGTAGACTTCCCCTTCTGGGGATGCCAAATCGGG @® Garcinia mangostana |CAATCCCGGAAAAGGGTCGGTGAGTTTTGG

Leu13Term CCAAAACTCACCGACCCTTTTCCGGGATTGCCCGATTTGGCATCC | 2090

TTG-TAG CCAGAAGGGGAAGTCTACGGAAAGAATGATGACGTGGCGGCAGT

AGCAACCATTGTGGCCACAATTAACCAATCAG

CTTTCCGTAGACTTCCC 2091

GGGAAGTCTACGGAAAG 2092

Reduced palmitate ATGGTTGCTACTGCCGCCACGTCATCATTCTTTCCGTTGACTTCCC | 2093

Acyl-ACP-thioesterase {CTTCTGGGGATGCCTAATCGGGCAATCCCGGAAAAGGGTCGGTG

Garcinia mangostana |AGTTTTGGGTCAATGAAGTCGAAATCCGCGG

Lys21Term CCGCGGATTTCGACTTCATTGACCCAAAACTCACCGACCCTTTTCC | 2094

AAA-TAA GGGATTGCCCGATTAGGCATCCCCAGAAGGGGAAGTCAACGGAA

AGAATGATCGACGTGGCGGCAGTAGCAACCAT

GGGATGCCTAATCGGGC | 2095

GCCCGATTAGGCATCCC | 2096 ® Reduced palmitate A AGCACGAAA AR AAAAACCA 09 : Acyl-ACP-thicesterase |TACTGCTGTGACATAGGCGTTTTTCCCAGTCACTTCTTCACCTGAC

Gossypium hirsutum |TCCTCTGACTCGAAAAACAAGAAGCTCGG

Ser8Term CCGAGCTTCTTGTTTTTCGAGTCAGAGGAGTCAGGTGAAGAAGTG | 2098

TCG-TAG ACTGGGAAAAACGCCTATGTCACAGCAGTAGCAACCATGGTTTTTA

AAAACAACTTCAATTTCGTGCTGAAATCCC

TGTGACATAGGCGTTTT 2099

AAAACGCCTATGTCACA 2100 - Reduced palmitate TGTTTTTAAAAACCATGGTTGCTACTGCTGTGACATCGGCGTTTTT | 2101

Acyl-ACP-thicesterase |CCCAGTCACTTCTTGACCTGACTCCTCTGACTCGAAAAACAAGAAG

Gossypium hirsutum |CTCGGAAGCATCAAGTCGAAGCCATCGGT

Ser16Term ACCGATGGCTTCGACTTGATGCTTCCGAGCTTCTTGTTTTTCGAGT | 2102

TCA-TGA CAGAGGAGTCAGGTCAAGAAGTGACTGGGAAAAACGCCGATGTCA

CAGCAGTAGCAACCATGGTTTTTAAAAACA

1112/01 01:47 pm 03137.009 — [NY]783559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos } Alteration I—

CACTTCTTGACCTGACT 2103

AGTCAGGTCAAGAAGTG 2104

Reduced palmitate TTGCTACTGCTGTGACATCGGCGTTTTTCCCAGTCACTTCTTCACC | 2105

Acyl-ACP-thioesterase |TGACTCCTCTGACTAGAAAAACAAGAAGCTCGGAAGCATCAAGTC

Gossypium hirsutum |GAAGCCATCGGTTTCTTCTGGAAGTTTGCA

Ser22Term TGCAAACTTCCAGAAGAAACCGATGGCTTCGACTTGATGCTTCCG | 2106

TCG-TAG AGCTTCTTGTTTTTCTAGTCAGAGGAGTCAGGTGAAGAAGTGACTG

GGAAAAACGCCGATGTCACAGCAGTAGCAA

CTCTGACTAGAAAAACA 2107 9 TGTTTTTCIAGTCAGAG 2108

Reduced palmitate GCTACTGCTGTGACATCGGCGTTTTTCCCAGTCACTTCTTCACCTG | 2109

Acyl-ACP-thioesterase |ACTCCTCTGACTCGTAAAACAAGAAGCTCGGAAGCATCAAGTCGA

Gossypium hirsutum |AGCCATCGGTTTCTTCTGGAAGTTTGCAAG

Lys23Term CTTGCAAACTTCCAGAAGAAACCGATGGCTTCGACTTGATGCTTCC | 2110

AAA-TAA GAGCTTCTTGTTTTACGAGTCAGAGGAGTCAGGTGAAGAAGTGAC

TGGGAAAAACGCCGATGTCACAGCAGTAGC

CTGACTCGTAAAACAAG 2111

CTTGTTTTACGAGTCAG 2112

Acyl-ACP-thioesterase |CATTCTTCCCCGTGTAGTCCCCGGTCACCTCCTCTAGACCAGGAA

Cuphea hookeriana | AGCCCGGAAATGGGTCATCGAGCTTCAGCCC

Ser14Term GGGCTGAAGCTCGATGACCCATTTCCGGGCTTTCCTGGTCTAGAG | 2114

TCG-TAG GAGGTGACCGGGGACTACACGGGGAAGAATGCAGAGCTTGCAGC

GGTAGCCACCATTGTCTTTCAACGAGCGGGAG

® CCCCGTGTAGTCCCCGG 3115

CCGGGGACTACACGGGG 2116

Acyl-ACP-thioesterase | CCGGTCACCTCCTCTTGACCAGGAAAGCCCGGAAATGGGTCATCG

Cuphea hookeriana |AGCTTCAGCCCCATCAAGCCCAAATTTGTCG

Arg21Term CGACAAATTTGGGCTTGATGGGGCTGAAGCTCGATGACCCATTTC | 2118

AGA-TGA CGGGCTTTCCTGGTCAAGAGGAGGTGACCGGGGACGACACGGEG

GAAGAATGCAGAGCTTGCAGCGGTAGCCACCAT

CCTCCTCTIGACCAGGA 2119

TCCTGGTCAAGAGGAGG 2120 11/12/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Reduced palmitate GCTACCGCTGCAAGCTCTGCATTCTTCCCCGTGTCGTCCCCGGTC | 2121

Acyl-ACP-thioesterase |ACCTCCTCTAGACCATGAAAGCCCGGAAATGGGTCATCGAGCTTC

Cuphea hookeriana | AGCCCCATCAAGCCCAAATTTGTCGCCAATG

Gly23Term CATTGGCGACAAATTTGGGCTTGATGGGGCTGAAGCTCGATGACC | 2122

GGA-TGA CATTTCCGGGCTTTCATGGTCTAGAGGAGGTGACCGGGGACGAC

ACGGGGAAGAATGCAGAGCTTGCAGCGGTAGC

CTAGACCATGAAAGCCC 2123

GGGCTTTCATGGTCTAG 2124

Reduced palmitate ACCGCTGCAAGCTCTGCATTCTTCCCCGTGTCGTCCCCGGTCACC | 2125

Acyl-ACP-thioesterase |TCCTCTAGACCAGGATAGCCCGGAAATGGGTCATCGAGCTTCAGC ® Cuphea hookeriana | CCCATCAAGCCCAAATTTGTCGCCAATGGCG

Lys24Term CGCCATTGGCGACAAATTTGGGCTTGATGGGGCTGAAGCTCGATG | 2126

AAG-TAG ACCCATTTCCGGGCTATCCTGGTCTAGAGGAGGTGACCGGGGAC

GACACGGGGAAGAATGCAGAGCTTGCAGCGGT

GACCAGGATAGCCCGGA

TCCGGGCTATCCTGGTC 2128

Reduced palmitate 4 AA A A J

Acyl-ACP-thioesterase |ACCTCCTCTAGGCCGTGAAAGCTCGGAAATGGGTCATCGAGCTTG CT

Cuphea lanceolata AGCCCCCTCAAGCCCAAATTTGTCGCCAATG

Gly23Term CATTGGCGACAAATTTGGGCTTGAGGGGGCTCAAGCTCGATGACC | 2130

GGA-TGA CATTTCCGAGCTTTCACGGCCTAGAGGAGGTGTCCGGGGACGGC

AGGGGGAAGAATGCAGAACTTGCAGCGGTGGC

CTAGGCCGTGAAAGCTC 2131

GAGCTTTCACGGCCTAG

® Reduced palmitate ACCGCTGCAAGTTCTGCATTCTTCCCCCTGCCGTCCCCGGACACC | 2133

Acyl-ACP-thioesterase |TCCTCTAGGCCGGGATAGCTCGGAAATGGGTCATCGAGCTTGAGC

Cuphea lanceolata CCCCTCAAGCCCAAATTTGTCGCCAATGCCG

Lys24Term CGGCATTGGCGACAAATTTGGGCTTGAGGGGGCTCAAGCTCGAT | 2134

AAG-TAG GACCCATTTCCGAGCTATCCCGGCCTAGAGGAGGTGTCCGGGGA

CGGCAGGGGGAAGAATGCAGAACTTGCAGCGGT

GGCCGGGATAGCTCGGA 2135

TCCGAGCTATCCCGGCC 2136 : Reduced palmitate GCAAGTTCTGCATTCTTCCCCCTGCCGTCCCCGGACACCTCCTCT | 2137

Acyl-ACP-thioesterase |AGGCCGGGAAAGCTCTGAAATGGGTCATCGAGCTTGAGCCCCCT

Cuphea lanceolata CAAGCCCAAATTTGTCGCCAATGCCGGGTTGA

Gly26Term TCAACCCGGCATTGGCGACAAATTTGGGCTTGAGGGGGCTCAAGC | 2138

GGA-TGA TCGATGACCCATTTCAGAGCTTTCCCGGCCTAGAGGAGGTGTCCG

GGGACGGCAGGGGGAAGAATGCAGAACTTGC

1112101 01:47 pm 03137.008 — [NY]793659.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

GAAAGCTCIGAAATGGG 2139

CCCATTTCAGAGCTTTC 2140

Reduced palmitate CATTCTTCCCCCTGCCGTCCCCGGACACCTCCTCTAGGCCGGGAA | 2141

Acyl-ACP-thioesterase |[AGCTCGGAAATGGGTGATCGAGCTTGAGCCCCCTCAAGCCCAAAT

Cuphea lanceolata TTGTCGCCAATGCCGGGTTGAAGGTTAAGGC

Ser29Term GCCTTAACCTTCAACCCGGCATTGGCGACAAATTTGGGCTTGAGG | 2142

TCA-TGA GGGCTCAAGCTCGATCACCCATTTCCGAGCTTTCCCGGCCTAGAG

GAGGTGTCCGGGGACGGCAGGGGGAAGAATG

AAATGGGTGATCGAGCT 2143 [ AGCTCGATCACCCATTT

Acyl-ACP-thicesterase |GAGTGCTACTGCGTAGCTGTTTCCGGTTTCTTCCCCAAAACCTCAC

Helianthus annuus TCTGGAGCCAAGACATCTGATAAGCTTGG

Ser8Term CCAAGCTTATCAGATGTCTTGGCTCCAGAGTGAGGTTTTGGGGAA | 2146

TCG-TAG GAAACCGGAAACAGCTACGCAGTAGCACTCATAGCTACCATGATT

AAAACACTTAAATGTCCGATCCACTTAAACG

TACTGCGTAGCTGTTTC

GAAACAGCTACGCAGTA 2148

Reduced palmitate AGTGTTTTAATCATGGTAGCTATGAGTGCTACTGCGTCGCTGTITC | 2149

Acyl-ACP-thicesterase |CGGTTTCTTCCCCATAACCTCACTCTGGAGCCAAGACATCTGATAA

Helianthus annuus GCTTGGAGGTGAACCAGGTAGTGTTGCTG

Lys17Term CAGCAACACTACCTGGTTCACCTCCAAGCTTATCAGATGTCTTGGC | 2150

AAA-TAA TCCAGAGTGAGGTTATGGGGAAGAAACCGGAAACAGCGACGCAG

TAGCACTCATAGCTACCATGATTAAAACACT

® CTTCCCCATAACCTCAC 2151

GTGAGGTTATGGGGAAG 2152

Reduced palmitate ATGGTAGCTATGAGTGCTACTGCGTCGCTGTTTCCGGTTTCTTCCC | 2153

Acyl-ACP-thicesterase |CAAAACCTCACTCTIGAGCCAAGACATCTGATAAGCTTGGAGGTG

Helianthus annuus AACCAGGTAGTGTTGCTGTGCGCGGAATCA

Gly21Term TGATTCCGCGCACAGCAACACTACCTGGTTCACCTCCAAGCTTATC | 2154

GGA-TGA AGATGTCTTGGCTCAAGAGTGAGGTTTTGGGGAAGAAACCGGAAA

CAGCGACGCAGTAGCACTCATAGCTACCAT

. CTCACTCTIGAGCCAAG 2155

CTTGGCTCAAGAGTGAG 2156 1112001 01:47 pm 03137.009 — [NY)793553.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos :

Reduced palmitate GCTATGAGTGCTACTGCGTCGCTGTTTCCGGTTTCTTCCCCAAAAC | 2157

Acyl-ACP-thioesterase |CTCACTCTGGAGCCTAGACATCTGATAAGCTTGGAGGTGAACCAG

Helianthus annuus GTAGTGTTGCTGTGCGCGGAATCAAGACAA

Lys23Term TTGTCTTGATTCCGCGCACAGCAACACTACCTGGTTCACCTCCAAG | 2158

AAG-TAG CTTATCAGATGTCTAGGCTCCAGAGTGAGGTTTTGGGGAAGAAAC

CGGAAACAGCGACGCAGTAGCACTCATAGC

CTGGAGCCTAGACATCT 2159

AGATGTCTAGGCTCCAG 2160

Reduced palmitate A AGCAA A A 3

Acyl-ACP-thioesterase [CCAGGAGCCTCCCCTTAACCTGGGAAGTTAGGCAACTGGTCATCG @® Cuphea palustris AGTTTGAGCCCTTCCTTGAAGCCCAAGTCAA

Lys21Term TTGACTTGGGCTTCAAGGAAGGGCTCAAACTCGATGACCAGTTGC | 2162

AAA-TAA CTAACTTCCCAGGTTAAGGGGAGGCTCCTGGGGATGGAACAGGG

AAGCATGCAGAACTTGCTGCAGCAGCCACCAT

CCTCCCCTTAACCTGGG 2163

CCCAGGTTAAGGGGAGG EZ

Reduced palmitate GCTGCAGCAAGTTCTGCATGCTTCCCTGTTCCATCCCCAGGAGCC | 2165

Acyl-ACP-thioesterase |[TCCCCTAAACCTGGGTAGTTAGGCAACTGGTCATCGAGTTTGAGC

Cuphea palustris CCTTCCTTGAAGCCCAAGTCAATCCCCAATG

Lys24Term CATTGGGGATTGACTTGGGCTTCAAGGAAGGGCTCAAACTCGATG | 2166

AAG-TAG ACCAGTTGCCTAACTACCCAGGTTTAGGGGAGGCTCCTGGGGATG

GAACAGGGAAGCATGCAGAACTTGCTGCAGC

AACCTGGGTAGTTAGGC 2167

GCCTAACTACCCAGGTT

@® [Reducedpamiate |TGOATGGTTCCCIGTICOATCCCOAGGAGCCTCOCCTARCETGS | 2168

Acyl-ACP-thioesterase [GAAGTTAGGCAACTGATCATCGAGTTTGAGCCCTTCCTTGAAGCC

Cuphea palustris CAAGTCAATCCCCAATGGCGGATTTCAGGTT

Trp28Term AACCTGAAATCCGCCATTGGGGATTGACTTGGGCTTCAAGGAAGG | 2170

TGG-TGA GCTCAAACTCGATGATCAGTTGCCTAACTTCCCAGGTTTAGGGGA

GGCTCCTGGGGATGGAACAGGGAAGCATGCA

GGCAACTGATCATCGAG 2171

CTCGATGAICAGTTGCC 2172

Acyl-ACP-thioesterase |[AGTTAGGCAACTGGTGATCGAGTTTGAGCCCTTCCTTGAAGCCCA

Cuphea palustris AGTCAATCCCCAATGGCGGATTTCAGGTTAA

Ser29Term TTAACCTGAAATCCGCCATTGGGGATTGACTTGGGCTTCAAGGAA | 2174

TCA-TGA GGGCTCAAACTCGATCACCAGTTGCCTAACTTCCCAGGTTTAGGG

GAGGCTCCTGGGGATGGAACAGGGAAGCATG

11/12/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration i ES—

CAACTGGTGATCGAGTT 2175

AACTCGATCACCAGTTG 2176

Acyl-ACP-thicesterase |CCGGGAATCTCCCCTTAACCCGGGAAGTTCGGTAATGGTGGCTTT

Cuphea hookeriana CAGGTTAAGGCAAACGCCAATGCCCATCCTA

Lys21Term TAGGATGGGCATTGGCGTTTGCCTTAACCTGAAAGCCACCATTAC | 2178

AAA-TAA CGAACTTCCCGGGTTAAGGGGAGATTCCCGGGGTTGGAACGGAG

AAGAATGCAGAACTTGCTGCGGCAGCCACCAT

TCTCCCCTTAACCCGGG 2179 ® CCCGGGTTAAGGGGAGA 2180

Acyl-ACP-thicesterase |TCCCCTAAACCCGGGTAGTTCGGTAATGGTGGCTTTCAGGTTAAG

Cuphea hookeriana | GCAAACGCCAATGCCCATCCTAGTCTAAAGT

Lys24Term ACTTTAGACTAGGATGGGCATTGGCGTTTGCCTTAACCTGAAAGC | 2182

AAG-TAG CACCATTACCGAACTACCCGGGTTTAGGGGAGATTCCCGGGGTTG

GAACGGAGAAGAATGCAGAACTTGCTGCGGC

AACCCGGGTAGTTCGGT 2183

ACCGAACTACCCGGGTT

Reduced palmitate TTCTCCGTTCCAACCCCGGGAATCTCCCCTAAACCCGGGAAGTTC | 2185

Acyl-ACP-thicesterase |GGTAATGGTGGCTTTTAGGTTAAGGCAAACGCCAATGCCCATCCT

Cuphea hookeriana |AGTCTAAAGTCTGGCAGCCTCGAGACTGAAG

GIn31Term CTTCAGTCTCGAGGCTGCCAGACTTTAGACTAGGATGGGCATTGG | 2186

CAG-TAG CGTTTGCCTTAACCTAAAAGCCACCATTACCGAACTTCCCGGGTTT

AGGGGAGATTCCCGGGGTTGCAACGGAGAA

® GTGGCTTTTAGGTTAAG 2187

CTTAACCTAAAAGCCAC 2188

Acyl-ACP-thicesterase |GGTGGCTTTCAGGTTTAGGCAAACGCCAATGCCCATCCTAGTCTA

Cuphea hookeriana |AAGTCTGGCAGCCTCGAGACTGAAGATGACA

Lys33Term TGTCATCTTCAGTCTCGAGGCTGCCAGACTTTAGACTAGGATGGG | 2190

AAG-TAG CATTGGCGTTTGCCTAAACCTGAAAGCCACCATTACCGAACTTCCC

GGGTTTAGGGGAGATTCCCGGGGTTGGAAC

: TTCAGGTTTAGGCAAAC 2191

GTTTGCCTAAACCTGAA

1112/01 01:47 pm 03137008 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Reduced palmitate A AA AA i ATAA ACAGA J

Acyl-ACP-thioesterase |TCTCGCATTCTCATTAACCGGATCCGGCACACCGGAGAACCGTCT

Brassica rapa CCTCCGTGTCGTGCTCTCATCTGAGGAAAC

Gin21Term GTTTCCTCAGATGAGAGCACGACACGGAGGAGACGGTTCTCCGGT| 2194

CAA-TAA GTGCCGGATCCGGTTAATGAGAATGCGAGAAGAGGGTCTGTAACT

TATCAGTCGCATTACACGAGAGCTTCAACAT

ATTCTCATTAACCGGAT 2195

ATCCGGTTAATGAGAAT

Reduced palmitate GCGACTGATAAGTTACAGACCCTCTTCTCGCATTCTCATCAACCGG | 2197

Acyl-ACP-thioesterase |ATCCGGCACACCGGIGAACCGTCTCCTCCGTGTCGTGCTCTCATC @ Brassica rapa TGAGGAAACCGGTTCTCGATCCTTTGCGAG

Arg28Term CTCGCAAAGGATCGAGAACCGGTTTCCTCAGATGAGAGCACGACA | 2198

AGA-TGA CGGAGGAGACGGTTCACCGGTGTGCCGGATCCGGTTGATGAGAA

TGCGAGAAGAGGGTCTGTAACTTATCAGTCGC

CACACCGGTIGAACCGTC 2199

GACGGTTCACCGGTGTG 2200

Reduced palmitate CCCTCTTCTCGCATTCTCATCAACCGGATCCGGCACACCGGAGAA | 2201

Acyl-ACP-thioesterase |CCGTCTCCTCCGTGTAGTGCTCTCATCTGAGGAAACCGGTTCTCG

Brassica rapa ATCCTTTGCGAGCGATCGTATCTGCTGATCA

Ser24Term TGATCAGCAGATACGATCGCTCGCAAAGGATCGAGAACCGGTTTC | 2202

TCG-TAG CTCAGATGAGAGCACTACACGGAGGAGACGGTTCTCCGGTGTGC

CGGATCCGGTTGATGAGAATGCGAGAAGAGGG

CTCCGTGTAGTGCTCTC 2203

GAGAGCACTACACGGAG 2204 ®

Acyl-ACP-thioesterase [CTCCTCCGTGTCGTGATCTCATCTGAGGAAACCGGTTCTCGATCC

Brassica rapa TTTGCGAGCGATCGTATCTGCTGATCAAGGA

Cys25Term TCCTTGATCAGCAGATACGATCGCTCGCAAAGGATCGAGAACCGG | 2206

TGC-TGA TTTCCTCAGATGAGATCACGACACGGAGGAGACGGTTCTCCGGTG

TGCCGGATCCGGTTGATGAGAATGCGAGAAG

GTGTCGTGATCTCATCT 2207

AGATGAGATCACGACAC 2208 .

Acyl-ACP-thioesterase | GCATCAAAAATGTAGAAGCTTTCGTGTAATGTGACTAACAACTTAC CT

Brassica napus ACACCTTCTCCTTCTTCTCCGATTCCTC

Leu2Term GAGGAATCGGAGAAGAAGGAGAAGGTGTGTAAGTTGTTAGTCACA | 2210

TTG-TAG TTACACGAAAGCTTCTACATTTTTGATGCCCTTTTTTTTTTATGGTTC

CTGAGGTTTTGGTTTATAGAAGAAGAAT

1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ) Alteration _— — — ___———— — —_— —————— ———=

AAAAATGTAGAAGCTTT 2211

AAAGCTTCTACATTTTT 2212

Reduced palmitate TCTTCTTCTATAAACCAAAACCTCAGGAACCATAAAAAAAAAAGGG | 2213

Acyl-ACP-thioesterase |CATCAAAAATGTTGTAGCTTTCGTGTAATGTGACTAACAACTTACAC

Brassica napus ACCTTCTCCTTCTTCTCCGATTCCTCCC

Lys3Term GGGAGGAATCGGAGAAGAAGGAGAAGGTGTGTAAGTTGTTAGTCA| 2214

AAG-TAG CATTACACGAAAGCTACAACATTTTTGATGCCCTTTTTTTTTTATGG

TTCCTGAGGTTTTGGTTTATAGAAGAAGA

AAATGTTGTAGCTTTCG 2215 ® CGAAAGCTACAACATTT 2216 —_— | <£'9

Reduced palmitate CTATAAACCAAAACCTCAGGAACCATAAAAAAAAAAGGGCATCAAA | 2217

Acyl-ACP-thioesterase |AATGTTGAAGCTTTAGTGTAATGTGACTAACAACTTACACACCTTCT

Brassica napus CCTTCTTCTCCGATTCCTCCCTTTTCAT

Ser5Term ATGAAAAGGGAGGAATCGGAGAAGAAGGAGAAGGTGTGTAAGTT | 2218

TCG-TAG GTTAGTCACATTACACTAAAGCTTCAACATTTTTGATGCCCTTTTTI

TTTTATGGTTCCTGAGGTTTTGGTTTATAG

GAAGCTTTAGTGTAATG 2219

CATTACACTAAAGCTTC

AAAACCTCAGGAACCATAAAAAAAAAAGGGCATCAAAAATG | 2221

Acyl-ACP-thicesterase | TTGAAGCTTTCGTGAAATGTGACTAACAACTTACACACCTTCTCCTT

Brassica napus CTTCTCCGATTCCTCCCTTTTCATCCCG

Cys6Term CGGGATGAAAAGGGAGGAATCGGAGAAGAAGGAGAAGGTGTGTA | 2222

TGT-TGA AGTTGTTAGTCACATTICACGAAAGCTTCAACATTTTTGATGCCCTT

TTTTTTTTATGGTTCCTGAGGTTTTGGTTT

® CTTTCGTGAAATGTGAC 2223

GTCACATTTICACGAAAG 2224 = | eect 1112101 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Table 23

Oligonucleotides to produce plants with increased stearate

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration _

Increased stearate GGGAGAGCTCTAGCTCTGTAGAAAAGAAGGATTCATTCATCATATC | 2225 stearoyl-ACP CAGAAATGGCTCTATAGTTTAACCCTTTGGTGGCATCTCAGCCTTA desaturase CAAATTCCCTTCCTCGACTCGTCCGCCAA

Arabidopsis thaliana | TTGGCGGACGAGTCGAGGAAGGGAATTTGTAAGGCTGAGATGCC | 2226

Lys4Term ACCAAAGGGTTAAACTATAGAGCCATTTCTGGATATGATGAATGAA

AAG-TAG TCCTTCTTTTCTACAGAGCTAGAGCTCTCCC

TGGCTCTATAGTTTAAC 2227 ® GTTAAACTATAGAGCCA 2228

Increased stearate CTCTGTAGAAAAGAAGGATTCATTCATCATATCCAGAAATGGCTCT | 2229 stearoyl-ACP AAAGTTTAACCCTTAGGTGGCATCTCAGCCTTACAAATTCCCTTCC desaturase TCGACTCGTCCGCCAACTCCTTCTTTCAG

Arabidopsis thaliana |CTGAAAGAAGGAGTTGGCGGACGAGTCGAGGAAGGGAATTTGTA | 2230

Leu8Term AGGCTGAGATGCCACCTAAGGGTTAAACTTTAGAGCCATTTCTGG

TTG-TAG ATATGATGAATGAATCCTTCTTTTCTACAGAG

TAACCCTTAGGTGGCAT 2231

ATGCCACCTAAGGGTTA 2232

Increased stearate AGAAGGATTCATTCATCATATCCAGAAATGGCTCTAAAGTTTAACC | 2233 stearoyl-ACP CTTTGGTGGCATCTTAGCCTTACAAATTCCCTTCCTCGACTCGTCC desaturase GCCAACTCCTTCTTTCAGATCTCCCAAGT

Arabidopsis thaliana | ACTTGGGAGATCTGAAAGAAGGAGTTGGCGGACGAGTCGAGGAA | 2234

Gin12Term GGGAATTTGTAAGGCTAAGATGCCACCAAAGGGTTAAACTTTAGA ® CAG-TAG GCCATTTCTGGATATGATGAATGAATCCTTCT

TGGCATCTTAGCCTTAC 2235

GTAAGGCTAAGATGCCA 2236

Increased stearate TCATTCATCATATCCAGAAATGGCTCTAAAGTTTAACCCTTTGGTG | 2237 stearoyl-ACP GCATCTCAGCCTTAGAAATTCCCTTCCTCGACTCGTCCGCCAACTC desaturase CTTCTTTCAGATCTCCCAAGTTCCTCTGC

Arabidopsis thaliana |GCAGAGGAACTTGGGAGATCTGAAAGAAGGAGTTGGCGGACGAG | 2238

Phe14Term TCGAGGAAGGGAATTTCTAAGGCTGAGATGCCACCAAAGGGTTAA : TAC-TAG ACTTTAGAGCCATTTCTGGATATGATGAATGA

CAGCCTTAGAAATTCCC 2239 : GGGAATTTCTAAGGCTG 2240 1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration ncreased stearate AGA AAAGAACA AAA A AAAAA 4 stearoyl-ACP AAAGAAAATGGCATAGAAGCTTAACCCTTTGGCATCTCAGCCTTAC desaturase AAACTCCCTTCCTCGGCTCGTCCGCCAAT

Brassica napus ATTGGCGGACGAGCCGAGGAAGGGAGTTTGTAAGGCTGAGATGC | 2242

Leu3Term CAAAGGGTTAAGCTTCTATGCCATTTTCTTTITTTTGATACGAGGTT

TTG-TAG TGATGTTCTTTCAGACACGAGCGAGCTCTC

AATGGCATAGAAGCTTA 2243

TAAGCTTCTATGCCATT

Increased stearate |GAGCTCGCTCGTGTCTGAAAGAACATCAAACCTCGTATCAAAAAAA | 2245 stearoyl-ACP AGAAAATGGCATTGTAGCTTAACCCTTTGGCATCTCAGCCTTACAA ® desaturase ACTCCCTTCCTCGGCTCGTCCGCCAATCT

Brassica napus AGATTGGCGGACGAGCCGAGGAAGGGAGTTTGTAAGGCTGAGAT | 2246

Lys4Term GCCAAAGGGTTAAGCTACAATGCCATTTTCTTTTTTTTGATACGAG

AAG-TAG GTTTGATGTTCTTTCAGACACGAGCGAGCTC

TGGCATTGTAGCTTAAC 2247

GTTAAGCTACAATGCCA 2248

Increased stearate ~~ [TCTGAAAGAACATCAAACCTCGTATCAAAAAAAAGAAAATGGCATT | 2249 stearoyl-ACP GAAGCTTAACCCTTAGGCATCTCAGCCTTACAAACTCCCTTCCTCG desaturase GCTCGTCCGCCAATCTCTACTCTCAGATC

Brassica napus GATCTGAGAGTAGAGATTGGCGGACGAGCCGAGGAAGGGAGTTT | 2250

Leu8Term GTAAGGCTGAGATGCCTAAGGGTTAAGCTTCAATGCCATTTTCTTT

TTG-TAG TTTTTGATACGAGGTTTGATGTTCTTTCAGA

TAACCCTTAGGCATCTC 2251

GAGATGCCTAAGGGTTA 2252 ® AAAAAAAAGAAAATGGCATTGAAGCTTA | 2253 stearoyl-ACP ACCCTTTGGCATCTTAGCCTTACAAACTCCCTTCCTCGGCTCGTCC desaturase GCCAATCTCTACTCTCAGATCTCCCAAGT

Brassica napus ACTTGGGAGATCTGAGAGTAGAGATTGGCGGACGAGCCGAGGAA | 2254

Gin11Term GGGAGTTTGTAAGGCTAAGATGCCAAAGGGTTAAGCTTCAATGCC

CAG-TAG ATTTTCTTTTTTTTGATACGAGGTTTGATGTT

TGGCATCTTAGCCTTAC 2255

GTAAGGCTAAGATGCCA 2256 ncreased sfearale AA AAAAGAAAAA AAGAAAAAAAL RA R7 x stearoyl-ACP ATCCTTTCCTTTCTTAAACCCAAAAGTTACCTTCTTTCGCTCTTCCA desaturase CCAATGGCCAGTACCAGATCTCCTAAGT : Ricinus communis ~~ [ACTTAGGAGATCTGGTACTGGCCATTGGTGGAAGAGCGAAAGAAG | 2258

GIn27Term GTAACTTTTGGGTTTAAGAAAGGAAAGGATTGAGCTTGAGAGCCAT

CAA-TAA TGTTITTTITCTTACCTTTTITCTTTTGGTT 1112/01 01:47 pm 03137.009 — (NY}793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos } Alteration a ———— ee ——

TCCTTTCTTAAACCCAA 2259

TTGGGTTTAAGAAAGGA 2260 increased stearate AAGAAAAAGGTAAGAAAAAAAACAATGGCTCTCAAGCTCAATCCTT | 2261 stearoyl-ACP TCCTTTCTCAAACCTAAAAGTTACCTTCTTTCGCTCTTCCACCAATG desaturase GCCAGTACCAGATCTCCTAAGTTCTACA

Ricinus communis TGTAGAACTTAGGAGATCTGGTACTGGCCATTGGTGGAAGAGCGA | 2262

Gin29Term AAGAAGGTAACTTTTAGGTTTGAGAAAGGAAAGGATTGAGCTTGA

CAA-TAA GAGCCATTGTTTTTTTTCTTACCTTTTTCTT

CTCAAACCTAAAAGTTA 2263 [ TAACTTTTAGGTTTGAG 2264

Increased stearate AAAAAGGTAAGAAAAAAAACAATGGCTCTCAAGCTCAATCCTTTCC | 2265 stearoyl-ACP TTTCTCAAACCCAATAGTTACCTTCTTTCGCTCTTCCACCAATGGCC desaturase AGTACCAGATCTCCTAAGTTCTACATGG

Ricinus communis CCATGTAGAACTTAGGAGATCTGGTACTGGCCATTGGTGGAAGAG | 2266

Lys30Term CGAAAGAAGGTAACTATTGGGTTTGAGAAAGGAAAGGATTGAGCT

AAG-TAG TGAGAGCCATTGTTTTTTITTCTTACCTTTTT

AAACCCAATAGTTACCT 2267

AGGTAACTATTGGGTTT 2268

Increased stearate TCTCAAACCCAAAAGTTACCTICTTTCGCTCTTCCACCAATGGCCA 2269 stearoyl-ACP GTACCAGATCTCCTTAGTTCTACATGGCCTCTACCCTCAAGTCTGG desaturase TTCTAAGGAAGTTGAGAATCTCAAGAAGC

Ricinus communis GCTTCTTGAGATTCTCAACTTCCTTAGAACCAGACTTGAGGGTAGA | 2270

Lysd6Term GGCCATGTAGAACTAAGGAGATCTGGTACTGGCCATTGGTGGAAG

AAG-TAG AGCGAAAGAAGGTAACTTTTGGGTTTGAGA ® GATCTCCTTAGTTCTAC 2271

GTAGAACTAAGGAGATC 2272 ncreased stearate A A AZ i ATCAA SAGACTIGAA T9073 stearoyl-ACP CCCTATCCCCACCTAAACCTTCTCCCTCCCCCAAATGGCCAGTCTC desaturase AGATCTCCCAGGTTCCGCATGGCCTCTA

Glycine max TAGAGGCCATGCGGAACCTGGGAGATCTGAGACTGGCCATTTGG | 2274

Gin11Term GGGAGGGAGAAGGTTTAGGTGGGGATAGGGTTCAGTCTCAGAGC

CAA-TAA CATTGATGAGTAAAGATTAAATGAATCAGAAGA ] TCCCCACCTIAAACCTTC 2275

GAAGGTTTAGGTGGGGA 2276 11/12/01 01:47 pm 03137.008 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos } Alteration

Ee 1

Increased stearate CTTTACTCATCAATGGCTCTGAGACTGAACCCTATCCCCACCCAAA | 2277 stearoyl-ACP CCTTCTCCCTCCCCIAAATGGCCAGTCTCAGATCTCCCAGGTTCC desaturase GCATGGCCTCTACCCTCCGCTCCGGTTCCA

Glycine max TGGAACCGGAGCGGAGGGTAGAGGCCATGCGGAACCTGGGAGAT | 2278

GIn17Term CTGAGACTGGCCATTTAGGGGAGGGAGAAGGTTTGGGTGGGGAT

CAA-TAA AGGGTTCAGTCTCAGAGCCATTGATGAGTAAAG

CCCTCCCCTAAATGGCC 2279

GGCCATTTAGGGGAGGG

Increased stearate GCTCTGAGACTGAACCCTATCCCCACCCAAACCTTCTCCCTCCCC 2281 stearoyl-ACP CAAATGGCCAGTCTCTGATCTCCCAGGTTCCGCATGGCCTCTACC ® desaturase CTCCGCTCCGGTTCCAAAGAGGTTGAAAATA

Glycine max TATTTTCAACCTCTTTGGAACCGGAGCGGAGGGTAGAGGCCATGC | 2282

Arg22Term GGAACCTGGGAGATCAGAGACTGGCCATTTGGGGGAGGGAGAAG

AGA-TGA GTTTGGGTGGGGATAGGGTTCAGTCTCAGAGC

GGGAGATCAGAGACTGG 2284

Increased stearate CAAATGGCCAGTCTCAGATCTCCCAGGTTCCGCATGGCCTCTACC | 2285 stearoyl-ACP CTCCGCTCCGGTTCCTAAGAGGTTGAAAATATTAAGAAGCCATTCA desaturase CTCCTCCCAGAGAAGTGCATGTTCAAGTAA

Glycine max TTACTTGAACATGCACTTCTCTGGGAGGAGTGAATGGCTTCTTAAT | 2286

Lys37Term ATTTTCAACCTCTTAGGAACCGGAGCGGAGGGTAGAGGCCATGCG

AAA-TAA GAACCTGGGAGATCTGAGACTGGCCATTTG

CCGGTTCCTAAGAGGTT 2287

AACCTCTTAGGAACCGG 2288 o ncreased stearate AA AA ) A J AA AA AA A AA AA A A HY stearoyl-ACP AATACGGCGACGTTTTAATCAGACCTGTACCGTTCATTCGCGTTTC desaturase CTCAACCGAAACCTCTCAGATCTCCCAAAT

Helianthus annuus ATTTGGGAGATCTGAGAGGTTTCGGTTGAGGAAACGCGAATGAAC | 2290

GIn11Term GGTACAGGTCTGATTAAAACGTCGCCGTATTGATGCGAATCGCCA

CAA-TAA TTGTTGATGTTGTTCTTGTGTGTGCTTGTTG

CGACGTTTTAATCAGAC 229A

GTCTGATTAAAACGTCG 2292

Increased stearate AAGCACACACAAGAACAACATCAACAATGGCGATTCGCATCAATACT 2293 stearoyl-ACP GGCGACGTTTCAATGAGACCTGTACCGTTCATTCGCGTTTCCTCAA desaturase CCGAAACCTCTCAGATCTCCCAAATTCGC

Helianthus annuus GCGAATTTGGGAGATCTGAGAGGTTTCGGTTGAGGAAACGCGAAT | 2294

Ser12Term GAACGGTACAGGTCTCATTGAAACGTCGCCGTATTGATGCGAATC

TCA-TGA GCCATTGTTGATGTTIGTTICTTGTGTGTGCTT 11112101 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

GTTTCAATGAGACCTGT

: ACAGGTCTCATTGAAAC 2296

Increased stearate AAGAACAACATCAACAATGGCGATTCGCATCAATACGGCGACGTTT| 2297 stearoyl-ACP CAATCAGACCTGTAGCGTTCATTCGCGTTTCCTCAACCGAAACCTC desaturase TCAGATCTCCCAAATTCGCCATGGCTTCC

Helianthus annuus GGAAGCCATGGCGAATTTGGGAGATCTGAGAGGTTTCGGTTGAGG| 2298

Tyr15Term AAACGCGAATGAACGCTACAGGTCTGATTGAAACGTCGCCGTATT

TAC-TAG GATGCGAATCGCCATTGTTGATGTTGTTCTT

GACCTGTAGCGTTCATT 2299 ® AATGAACGCTACAGGTC 2300 stearoyl-ACP AGACCTGTACCGTTGATTCGCGTTTCCTCAACCGAAACCTCTCAGA desaturase TCTCCCAAATTCGCCATGGCTTCCACCAT

Helianthus annuus ATGGTGGAAGCCATGGCGAATTTGGGAGATCTGAGAGGTTTCGGT | 2302

Ser17Term TGAGGAAACGCGAATCAACGGTACAGGTCTGATTGAAACGTCGCC

TCA-TGA GTATTGATGCGAATCGCCATTGTTGATGTTG

GTACCGTIGATICGCGT ~~ | 2303

ACGCGAATCAACGGTAC 2304 stearoyl-ACP AACGATGGCGCTTIGAATGAGTCCGGTGACGCTTCAACGGGAGAT desaturase ATATCCTTCATACACTTTTCATCAATCGA

Helianthus annuus TCGATTGATGAAAAGTGTATGAAGGATATATCTCCCGTTGAAGCGT | 2306

Arg4Term CACCGGACTCATTCAAAGCGCCATCGTTGATGAAGAAGATGATGA

CGA-TGA TGTGTGTGTGATTGAGTGTGTGTTGTGTGT ® TGGCGCTTTGAATGAGT 2307

ACTCATTCAAAGCGCCA 2308 stearoyl-ACP GTCCGGTGACGCTTTAACGGGAGATATATCCTTCATACACTTTTCA desaturase TCAATCGAAAAATCTCAGATCTCCTAAAT

Helianthus annuus ATTTAGGAGATCTGAGATTTTTCGATTGATGAAAAGTGTATGAAGG | 2310

GIn11Term ATATATCTCCCGTTAAAGCGTCACCGGACTCATTCGAAGCGCCATC

CAA-TAA GTTGATGAAGAAGATGATGATGTGTGTGT

TGACGCTTTAACGGGAG 2311

CTCCCGTTAAAGCGTCA 2312 111201 01:47 pm 03137.009 — [NY]793550.1

AMENDED SHEET

28-12-2001 01939797 8 EE es CCE. EASE ee AES

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

Increased stearate ACATCATCATCTTCTTCATCAACGATGGCGCTTCGAATGAGTCCGG | 2313 stearoyl-ACP TGACGCTTCAACGGTAGATATATCCTTCATACACTTTTCATCAATCG desaturase AAAAATCTCAGATCTCCTAAATTCGCGA

Helianthus annuus TCGCGAATTTAGGAGATCTGAGATTTTTCGATTGATGAAAAGTGTA | 2314

Glu13Term TGAAGGATATATCTACCGTTGAAGCGTCACCGGACTCATTCGAAG

GAG-TAG CGCCATCGTTGATGAAGAAGATGATGATGT

TTCAACGGTAGATATAT 2315

ATATATCTACCGTTGAA 2316

Increased stearate ATCTTCTTCATCAACGATGGCGCTTCGAATGAGTCCGGTGACGCTT 2317 stearoyl-ACP CAACGGGAGATATAGCCTTCATACACTTTTCATCAATCGAAAAATC @ desaturase TCAGATCTCCTAAATTCGCGATGGCTTCC

Helianthus annuus GGAAGCCATCGCGAATTTAGGAGATCTGAGATTTTTCGATTGATGA | 2318

Tyr15Term AAAGTGTATGAAGGCTATATCTCCCGTTGAAGCGTCACCGGACTC

TAT-TAG ATTCGAAGCGCCATCGTTGATGAAGAAGAT

GAGATATAGCCTTCATA 2319

TATGAAGGCTATATCTC 2320 stearoyl-ACP ACAACAATGGCTCTCTAGCTCAACCCAGTCACCACCTTCCCTTCAA desaturase CACGCTCCCTCAACAACTTCTCCTCCAGAT

Linum usitatissimum |ATCTGGAGGAGAAGTTGTTGAGGGAGCGTGTTGAAGGGAAGGTG | 2322

Lys4Term GTGACTGGGTTGAGCTAGAGAGCCATTGTIGTTGAAGGTTITTCT

AAG-TAG GCGCTGTTGTTTGGGGGCAAGCTGGCTGAGTT

TGGCTCTCTAGCTCAAC 2323

GTTGAGCTAGAGAGCCA 2324 ® stearoyl-ACP TCACCACCTTCCCTTGAACACGCTCCCTCAACAACTTCTCCTCCAG desaturase ATCTCCTCGCACCTTTCTCATGGCTGCTTC : Linum usitatissimum |GAAGCAGCCATGAGAAAGGTGCGAGGAGATCTGGAGGAGAAGTT | 2326

Ser13Term GTTGAGGGAGCGTGTTCAAGGGAAGGTGGTGACTGGGTTGAGCT

TCA-TGA TGAGAGCCATTGTTGTTGAAGGTTTTITCTGCGC

CTTCCCTTGAACACGCT 2327

AGCGTGTTCAAGGGAAG 2328 .25 Increased stearate CTCAAGCTCAACCCAGTCACCACCTTCCCTTCAACACGCTCCCTCA! 2329 stearoyl-ACP ACAACTTCTCCTCCTIGATCTCCTCGCACCTTTCTCATGGCTGCTTC desaturase CACTTTCAATTCCACCTCCACCAAGTAAG

Linum usitatissimum |CTTACTTGGTGGAGGTGGAATTGAAAGTGGAAGCAGCCATGAGAA | 2330

Arg23Term AGGTGCGAGGAGATCAGGAGGAGAAGTTGTTGAGGGAGCGTGTT

AGA-TGA GAAGGGAAGGTGGTGACTGGGTTIGAGCTTGAG 1412/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797 — ee

Phenotype, Gene,

Plant & Targeted Altering Oligos } Alteration [fercercereareneer

AGGAGATCAGGAGGAGA 2332 stearoyl-ACP ATTCCACCTCCACCTAGTAAGCATCTCCTCCTCCTCGGAATCTCCG desaturase CCGATTTCTTTTAAGCGATTGATCGTAGA

Linum usitatissimum | TCTACGATCAATCGCTTAAAAGAAATCGGCGGAGATTCCGAGGAG | 2334

Lys41Term GAGGAGATGCTTACTAGGTGGAGGTGGAATTGAAAGTGGAAGCA

AAG-TAG GCCATGAGAAAGGTGCGAGGAGATCTGGAGGA

CCTCCACCTAGTAAGCA 2335 @ TGCTTACTAGGTGGAGG 2336 ncreased stearate i A AAA A ACAAGA stearoyl-ACP CCGATGCTCGTATCTIGATCTCACAGGGTTTTCATGGCTTCAACTAT desaturase TCATTCTCCTTCTATGGAGGTCGGAAAAG

Olea europaea CTTTTCCGACCTCCATAGAAGGAGAATGAATAGTTGAAGCCATGAA | 2338

Arg21Term AACCCTGTGAGATCAGATACGAGCATCGGGGAAGGAAGGCATCTT

AGA-TGA GTGGGGTGGAAAGCAAAGTTTCAGTGCCAT

CTCGTATCIGATCTCAC 2339

GTGAGATCAGATACGAG 2340

Increased stearate CCCACAAGATGCCTTCCTTCCCCGATGCTCGTATCAGATCTCACAG| 2341 stearoyl-ACP GGTTTTCATGGCTTGAACTATTCATTCTCCTTCTATGGAGGTCGGA desaturase AAAGTTAAAAAGCCTTTCACGCCTCCACG

Olea europaea CGTGGAGGCGTGAAAGGCTTTTTAACTTTTCCGACCTCCATAGAA 2342

Ser29Term GGAGAATGAATAGTTCAAGCCATGAAAACCCTGTGAGATCTGATAC

TCA-TGA GAGCATCGGGGAAGGAAGGCATCTTGTGGG @® CATGGCTTGAACTATTC 2343

GAATAGTTCAAGCCATG 2344

Increased stearate GATGCTCGTATCAGATCTCACAGGGTTTTCATGGCTTCAACTATTC | 2345 stearoyl-ACP ATTCTCCTTCTATGTAGGTCGGAAAAGTTAAAAAGCCTTTCACGCC desaturase TCCACGAGAGGTACATGTTCAAGTAACCC

Olea europaea GGGTTACTTGAACATGTACCTCTCGTGGAGGCGTGAAAGGCTTTT | 2346

Glu37Term TAACTTTTCCGACCTACATAGAAGGAGAATGAATAGTTGAAGCCAT

GAG-TAG GAAAACCCTGTGAGATCTGATACGAGCATC : CTTCTATGTAGGTCGGA 2347

TCCGACCTACATAGAAG 2348

IN——— Udit etd td Wat 1112/01 01:47 pm 03137.009 — [NY)793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration stearoyl-ACP CTTCTATGGAGGTCTGAAAAGTTAAAAAGCCTTTCACGCCTCCACG desaturase AGAGGTACATGTTCAAGTAACCCATTCCT

Olea europaea AGGAATGGGTTACTTGAACATGTACCTCTCGTGGAGGCGTGAAAG | 2350

Gly39Term GCTTTTTAACTTTTCAGACCTCCATAGAAGGAGAATGAATAGTTGA

GGA-TGA AGCCATGAAAACCCTGTGAGATCTGATACG

TGGAGGTCTGAAAAGTT 2351

AACTTTTCAGACCTCCA 2352 ncreased stearate ATCA } . |stearoyl-ACP GAGAAATGGCACTGTAACTCAGTCCAGTCATGTTTCAATCTCAGAA @® desaturase GCTTCCATTTCTTGCCTCCTATCCGCCTT

Persea americana AAGGCGGATAGGAGGCAAGAAATGGAAGCTTCTGAGATTGAAACA | 2354

Lysd4Term TGACTGGACTGAGTTACAGTGCCATTTCTCCGTGCCTGATAGAGA

AAA-TAA GAGAGAGCAGAGGGGACGACAAAAACGAGAA

TGGCACTGIAACTCAGT 2355

ACTGAGTTACAGTGCCA 2356

Increased stearate CTGCTCTCTCTCTCTATCAGGCACGGAGAAATGGCACTGAAACTCA| 2357 stearoyl-ACP GTCCAGTCATGTTTTAATCTCAGAAGCTTCCATTTCTTGCCTCCTAT desaturase CCGCCTTCCAATCTCAGATCTCCGAGGG

Persea americana CCCTCGGAGATCTGAGATTGGAAGGCGGATAGGAGGCAAGAAAT | 2358

GIn11Term GGAAGCTTCTGAGATTAAAACATGACTGGACTGAGTTTCAGTGCC

CAA-TAA ATTTCTCCGTGCCTGATAGAGAGAGAGAGCAG

TCATGTTTTAATCTCAG 2359

CTGAGATTAAAACATGA 2360 @® Increased stearate TCTCTCTCTATCAGGCACGGAGAAATGGCACTGAAACTCAGTCCA | 2361 stearoyl-ACP GTCATGTTTCAATCTTAGAAGCTTCCATTTCTTGCCTCCTATCCGCC desaturase TTCCAATCTCAGATCTCCGAGGGTTTTCA

Persea americana TGAAAACCCTCGGAGATCTGAGATTGGAAGGCGGATAGGAGGCAA] 2362

Gln13Term GAAATGGAAGCTTCTAAGATTGAAACATGACTGGACTGAGTTTCAG

CAG-TAG TGCCATTTCTCCGTGCCTGATAGAGAGAGA

TTCAATCTTAGAAGCTT 2363

AAGCTTCTAAGATTGAA 2364 ‘25 Increased stearate CTCTCTATCAGGCACGGAGAAATGGCACTGAAACTCAGTCCAGTC | 2365 stearoyl-ACP ATGTTTCAATCTCAGTAGCTTCCATTTCTTGCCTCCTATCCGCCTTC desaturase CAATCTCAGATCTCCGAGGGTTTTCATGG

Persea americana CCATGAAAACCCTCGGAGATCTGAGATTGGAAGGCGGATAGGAG 2366

Lys14Term GCAAGAAATGGAAGCTACTGAGATTGAAACATGACTGGACTGAGT

AAG-TAG TTCAGTGCCATTTCTCCGTGCCTGATAGAGAG 11/12/01 01:47 pm 03137.009 — [NY]793558.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos , Alteration EIS ES—

AATCTCAGTAGCTTCCA 2367 : TGGAAGCTACTGAGATT 2368 ncreasea stearate AGA A OY stearoyl-ACP ACACCGCATCGCCGTAGTCCTGCGGCGGCGTGGCGCAGAGGAG desaturase GAGCAATGGGATGTCGAAGATGGTGGCCATGGCC

Oryza sativa GGCCATGGCCACCATCTTCGACATCCCATTGCTCCTCCTCTGCGC | 2370

Tyr12Term CACGCCGCCGCAGGACTACGGCGATGCGGTGTGGGACGCCGCE

TAC-TAG AACGCCATGAGCAGCGGCAGCGAGATCTCGGGG

TCGCCGTAGTCCTGCGG 2371 ® CCGCAGGACTACGGCGA 2372

Increased stearate CTGCTCATGGCGTTCGCGGCGTCCCACACCGCATCGCCGTACTCC| 2373 stearoyl-ACP TGCGGCGGCGTGGCGTAGAGGAGGAGCAATGGGATGTCGAAGAT desaturase GGTGGCCATGGCCTCCACCATCAACAGGGTCA

Oryza sativa TGACCCTGTTGATGGTGGAGGCCATGGCCACCATCTTCGACATCC | 2374

GIn19Term CATTGCTCCTCCTCTACGCCACGCCGCCGCAGGAGTACGGCGAT

CAG-TAG GCGGTGTGGGACGCCGCGAACGCCATGAGCAG

GCGTGGCGTAGAGGAGG 2375

CCTCCTCTACGCCACGC 2376

Increased stearate CCCACACCGCATCGCCGTACTCCTGCGGCGGCGTGGCGCAGAGG | 2377 stearoyl-ACP AGGAGCAATGGGATGTAGAAGATGGTGGCCATGGCCTCCACCAT desaturase CAACAGGGTCAAGACTGCTAAGAAGCCCTACAC

Oryza sativa GTGTAGGGCTTCTTAGCAGTCTTGACCCTGTTGATGGTGGAGGCC | 2378

Ser26Term ATGGCCACCATCTTCTACATCCCATTGCTCCTCCTCTGCGCCACGE

TCG-TAG CGCCGCAGGAGTACGGCGATGCGGTGTGGEG ® TGGGATGTAGAAGATGG 2379

CCATCTTCTACATCCCA

Increased stearate CACACCGCATCGCCGTACTCCTGCGGCGGCGTGGCGCAGAGGAG | 2381 stearoyl-ACP GAGCAATGGGATGTCGTAGATGGTGGCCATGGCCTCCACCATCAA desaturase CAGGGTCAAGACTGCTAAGAAGCCCTACACTC

Oryza sativa GAGTGTAGGGCTTCTTAGCAGTCTTGACCCTGTTGATGGTGGAGG | 2382

Lys27Term CCATGGCCACCATCTACGACATCCCATTGCTCCTCCTCTGCGCCA

AAG-TAG CGCCGCCGCAGGAGTACGGCGATGCGGTGTG : GGATGTCGTAGATGGTG 2383

CACCATCTACGACATCC 2384 1112001 01:47 pm 03137.009 — [NY]793553.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration ncreased stearafe A i ACCAACAGAAGCA AGGAGHA 3 stearoyl-ACP GAGAAGCAATGGCGTAGAAGCTTCACCACACGGCCTTCAATCCTT desaturase CCATGGCGGTTACCTCTTCGGGACTTCCTCG

Simmondsia chinensis |CGAGGAAGTCCCGAAGAGGTAACCGCCATGGAAGGATTGAAGGC | 2386

Leu3Term CGTGTGGTGAAGCTTCTACGCCATTGCTTCTCTCTCCTAAGTGCTT

TTG-TAG CTGTTGGTAACCGCTCAACCTAGAGAGAGAA

AATGGCGTAGAAGCTTC 2387

GAAGCTTCTACGCCATT 2388

Increased stearate CTCTCTCTAGGTTGAGCGGTTACCAACAGAAGCACTTAGGAGAGA | 2389 stearoyl-ACP GAAGCAATGGCGTTGTAGCTTCACCACACGGCCTTCAATCCTTCC ) desaturase ATGGCGGTTACCTCTTCGGGACTTCCTCGAT

Simmondsia chinensis |ATCGAGGAAGTCCCGAAGAGGTAACCGCCATGGAAGGATTGAAG 2390

Lysd4Term GCCGTGTGGTGAAGCTACAACGCCATTGCTTCTCTCTCCTAAGTG

AAG-TAG CTTCTGTTGGTAACCGCTCAACCTAGAGAGAG

TGGCGTTGTAGCTTCAC 2391

GTGAAGCTACAACGCCA 2392

Increased stearate AAGCAATGGCGTTGAAGCTTCACCACACGGCCTTCAATCCTTCCAT| 2393 stearoyl-ACP GGCGGTTACCTCTTAGGGACTTCCTCGATCGTATCACCTCAGATCT desaturase CACCGCGTTTTCATGGCTTCTTCTACAAT

Simmondsia chinensis |ATTGTAGAAGAAGCCATGAAAACGCGGTGAGATCTGAGGTGATAC | 2394

Ser19Term GATCGAGGAAGTCCCTAAGAGGTAACCGCCATGGAAGGATTGAAG

TCG-TAG GCCGTGTGGTGAAGCTTCAACGCCATTGCTT

TACCTCTTAGGGACTTC 2395

GAAGTCCCTIAAGAGGTA 2396

J Increased stearate GCAATGGCGTTGAAGCTTCACCACACGGCCTTCAATCCTTCCATG | 2397 stearoyl-ACP GCGGTTACCTCTTCGIGACTTCCTCGATCGTATCACCTCAGATCTC desaturase ACCGCGTTTTCATGGCTTCTTCTACAATTG

Simmondsia chinensis |CAATTGTAGAAGAAGCCATGAAAACGCGGTGAGATCTGAGGTGAT | 2398

Gly20Term ACGATCGAGGAAGTCACGAAGAGGTAACCGCCATGGAAGGATTG

GGA-TGA AAGGCCGTGTGGTGAAGCTTCAACGCCATTGC

CCTCTTCGIGACTTCCT 2399

AGGAAGTCACGAAGAGG 2400 ncreased stearate AA AA ACACCA i 20 stearoyl-ACP ATTGCCGTCTTTCTGACCTCGTCAAACGCCTTCTCGCAGATCTCCC desaturase AAATTCTTCATGGCTTCCACTCTCAGCAG

Spinacia oleracea CTGCTGAGAGTGGAAGCCATGAAGAATTTGGGAGATCTGCGAGAA | 2402

Ser21Term GGCGTTTGACGAGGTCAGAAAGACGGCAATCGACGACACTGAAAT

A-TGA GGTGTGGAAACGGGGTTGAGATTCAGAGCCA 141112001 01:47 pm 03137.009 — [NY}793559.1

AMENDED SHEET

28-12-2001 01939797 — ee _

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

GTCTITCTGACCICGIC 2403 i GACGAGGTCAGAAAGAC 2404

Increased stearate AATCTCAACCCCGTTTCCACACCATTTCAGTGTCGTCGATTGCCGT | 2405 stearoyl-ACP CTTTCTCACCTCGTTAAACGCCTTCTCGCAGATCTCCCAAATTCTT desaturase CATGGCTTCCACTCTCAGCAGCTCTTCTC

Spinacia oleracea GAGAAGAGCTGCTGAGAGTGGAAGCCATGAAGAATTTGGGAGATC | 2406

GIn24Term TGCGAGAAGGCGTTTAACGAGGTGAGAAAGACGGCAATCGACGA

CAA-TAA CACTGAAATGGTGTGGAAACGGGGTTGAGATT

CACCTCGTTAAACGCCT 2407

C AGGCGTTTAACGAGGTG 2408

Increased stearate TCCACACCATTTCAGTGTCGTCGATTGCCGTCTTTCTCACCTCGTC | 2409 stearoyl-ACP AAACGCCTTCTCGCTGATCTCCCAAATTCTTCATGGCTTCCACTCT desaturase CAGCAGCTCTTCTCCTAAGGAAGCGGAAA

Spinacia oleracea TTTCCGCTTCCTTAGGAGAAGAGCTGCTGAGAGTGGAAGCCATGA | 2410

Arg29Term AGAATTTGGGAGATCAGCGAGAAGGCGTTTGACGAGGTGAGAAA

AGA-TGA GACGGCAATCGACGACACTGAAATGGTGTGGA

CTTCTCGCIGATCTCCC

GGGAGATCAGCGAGAAG 2412

Increased stearate TTTCAGTGTCGTCGATTGCCGTCTTTCTCACCTCGTCAAACGCCTT | 2413 stearoyl-ACP CTCGCAGATCTCCCTAATTCTTCATGGCTTCCACTCTCAGCAGCTC desaturase TTCTCCTAAGGAAGCGGAAAGCCTGAAGA

Spinacia oleracea TCTTCAGGCTTTCCGCTTCCTTAGGAGAAGAGCTGCTGAGAGTGG | 2414 © |Lys32Term AAGCCATGAAGAATTAGGGAGATCTGCGAGAAGGCGTTTGACGAG

AAA-TAA GTGCAGAAAGACGGCAATCGACGACACTGAAA [ GATCTCCCTAATTCTTC 2475

GAAGAATTAGGGAGATC stearoyl-ACP CAATGGGGTGTCGTGAAAATCTCACAAAATGTTACCATTTCCTTGT desaturase TCTTCAGCCAGATCTGAGCGAGTTTTCAT

Solanum tuberosum |ATGAAAACTCGCTCAGATCTGGCTGAAGAACAAGGAAATGGTAAC | 2418

Leu10Term ATTTTGTGAGATTTTCACGACACCCCATTGATATTCAGTGCCATTGT

TTA-TGA TGATGCTCTGTTTTTCACCTCGACTATTT : GGTGTCGTGAAAATCTC 2419

GAGATTTTCACGACACC 2420 = | cY 11112101 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos :

Increased stearate ATAGTCGAGGTGAAAAACAGAGCATCAACAATGGCACTGAATATCA| 2421 stearoyl-ACP ATGGGGTGTCGTTATAATCTCACAAAATGTTACCATTTCCTTGTTCT desaturase TCAGCCAGATCTGAGCGAGTTTTCATGG

Solanum tuberosum |CCATGAAAACTCGCTCAGATCTGGCTGAAGAACAAGGAAATGGTA | 2422

Lys11Term ACATTTTGTGAGATTATAACGACACCCCATTGATATTCAGTGCCATT

AAA-TAA GTTGATGCTCTGTTTTTCACCTCGACTAT

TGTCGTTATAATCTCAC 2423

GTGAGATTATAACGACA 2424

Increased stearate GTGAAAAACAGAGCATCAACAATGGCACTGAATATCAATGGGGTG | 2425 stearoyl-ACP TCGTTAAAATCTCACTAAATGTTACCATTTCCTTGTTCTTCAGCCAG ® desaturase ATCTGAGCGAGTTTTCATGGCTTCAACCA

Solanum tuberosum | TGGTTGAAGCCATGAAAACTCGCTCAGATCTGGCTGAAGAACAAG | 2426

Lys14Term GAAATGGTAACATTTAGTGAGATTTTAACGACACCCCATTGATATT

AAA-TAA CAGTGCCATTGTTGATGCTCTGTTTTTCAC

AATCTCACTAAATGTTA 2427

TAACATTTAGTGAGATT 2428

Increased stearate ACAGAGCATCAACAATGGCACTGAATATCAATGGGGTGTCGTTAAA| 2429 stearoyl-ACP ATCTCACAAAATGTGACCATTTCCTTGTTCTTCAGCCAGATCTGAG desaturase CGAGTTTTCATGGCTTCAACCATTCATCG

Solanum tuberosum |CGATGAATGGTTGAAGCCATGAAAACTCGCTCAGATCTGGCTGAA | 2430

Leu16Term GAACAAGGAAATGGTCACATTTTGTGAGATTTTAACGACACCCCAT

TTA-TGA TGATATTCAGTGCCATTGTTGATGCTCTGT

CAAAATGTGACCATTTC 2431

GAAATGGTCACATTTTG 2432 @® ncreased stearate A AA AA ACAGAA 4 stearoyl-ACP TCCTTCTTCATCATGATCTTCTTCTTCTTCATCGTTCTCGCTTCCTC desaturase AAATGGCTAGCCTCAGATCTCCAAGGTT

Arachis hypogaea AACCTTGGAGATCTGAGGCTAGCCATTTGAGGAAGCGAGAACGAT | 2434

Ser21Term GAAGAAGAAGAAGATCATGATGAAGAAGGAGAGAGAAAGAGCTTC

TCA-TGA TGTGAAGGGTTAGGGTTCAGCCTCAGAGCCA

TTCATCATGATCTTCTT 2435

AAGAAGATCATGATGAA 2436 2 stearoyl-ACP TCTTCTTCTTCTTGATCGTTCTCGCTTCCTCAAATGGCTAGCCTCA desaturase GATCTCCAAGGTTCCGCATGGCCTCCAC

Arachis hypogaea GTGGAGGCCATGCGGAACCTTGGAGATCTGAGGCTAGCCATTTGA | 2438

Ser26Term GGAAGCGAGAACGATCAAGAAGAAGAAGATGATGATGAAGAAGGA

TCA-TGA GAGAGAAAGAGCTTCTGTGAAGGGTTAGGGT 11/12/01 01:47 pm 03137.009 — {NY]793550.

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

TTCTTCTTGATCGTTCT 2439 : AGAACGATCAAGAAGAA

Increased stearate CTAACCCTTCACAGAAGCTCTTTCTCTCTCCTICTTCATCATCATCT 2441 stearoyl-ACP TCTTCTTCTTCATAGTTCTCGCTTCCTCAAATGGCTAGCCTCAGAT desaturase CTCCAAGGTTCCGCATGGCCTCCACCCT

Arachis hypogaea AGGGTGGAGGCCATGCGGAACCTTGGAGATCTGAGGCTAGCCAT | 2442

Ser27Term TTGAGGAAGCGAGAACTATGAAGAAGAAGAAGATGATGATGAAGA i

TCG-TAG AGGAGAGAGAAAGAGCTTCTGTGAAGGGTTAG

TTCTTCATAGTTCTCGC 2443 9 GCGAGAACTATGAAGAA 2444

Increased stearate CTTCACAGAAGCTCTTTCTCTCTCCTTCTTCATCATCATCTTCTTCT | 2445 stearoyl-ACP TCTTCATCGTTCTAGCTTCCTCAAATGGCTAGCCTCAGATCTCCAA desaturase GGTTCCGCATGGCCTCCACCCTCCGCAC

Arachis hypogaea GTGCGGAGGGTGGAGGCCATGCGGAACCTTGGAGATCTGAGGCT | 2446

Ser29Term AGCCATTTGAGGAAGCTAGAACGATGAAGAAGAAGAAGATGATGA

TCG-TAG TGAAGAAGGAGAGAGAAAGAGCTTCTGTGAAG

ATCGTTCTAGCTTCCTC 2447

GAGGAAGCTAGAACGAT 2448 [Increased stearate AAA AAAA AAAA AAACCAGGAAZ AAACGAA 443 stearoyl-ACP AAGAAAAAATGGCTTAGAATTTTAATGCCATCGCCTCGAAATCTCA desaturase GAAGCTCCCTTGCTTTGCTCTTCCACCAAA

Gossypium hirsutum | TTTGGTGGAAGAGCAAAGCAAGGGAGCTTCTGAGATTTCGAGGCG | 2450

Leu3Term ATGGCATTAAAATTCTAAGCCATTTTTTCTTTTCGTTTGCCTTTCCT

TTG-TAG GGTTTGGGTTTTGGACGGCTTTTAACTTT @ AATGGCTTAGAATTTTA 2451

TAAAATTCTAAGCCATT 2452

Increased stearate CCCAAACCAGGAAAGGCAAACGAAAAGAAAAAATGGCTTTGAATTT| 2453 stearoyl-ACP TAATGCCATCGCCTAGAAATCTCAGAAGCTCCCTTGCTTTGCTCTT desaturase CCACCAAAGGCCACCCTTAGATCTCCCAA

Gossypium hirsutum | TTGGGAGATCTAAGGGTGGCCTTTGGTGGAAGAGCAAAGCAAGG | 2454

Ser1-Term GAGCTTCTGAGATTTCTAGGCGATGGCATTAAAATTCAAAGCCATT

TCG-TAG TTTTCTTTTCGTTTGCCTTTCCTGGTTTGGG } CATCGCCTAGAAATCTC 2455

GAGATTTCTAGGCGATG 2456 11/42/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration

Increased stearate CAAACCAGGAAAGGCAAACGAAAAGAAAAAATGGCTTTGAATTTTA | 2457 stearoyl-ACP ATGCCATCGCCTCGTAATCTCAGAAGCTCCCTTGCTTTGCTCTTCC desaturase ACCAAAGGCCACCCTTAGATCTCCCAAGT

Gossypium hirsutum |ACTTGGGAGATCTAAGGGTGGCCTTTGGTGGAAGAGCAAAGCAAG | 2458

Lys11Term GGAGCTTCTGAGATTACGAGGCGATGGCATTAAAATTCAAAGCCA

AAA-TAA TTITTTCTTTTCGTITGCCTITCCTGGTTTG

TCGCCTCGTAATCTCAG

CTGAGATTACGAGGCGA 2460

Increased stearate AGGAAAGGCAAACGAAAAGAAAAAATGGCTTTGAATTTTAATGCCA | 2461 stearoyl-ACP TCGCCTCGAAATCTTAGAAGCTCCCTTGCTTTGCTCTTCCACCAAA @® desaturase GGCCACCCTTAGATCTCCCAAGTTTTCCA

Gossypium hirsutum | TGGAAAACTTGGGAGATCTAAGGGTGGCCTTTGGTGGAAGAGCAA | 2462

GIn13Term AGCAAGGGAGCTTCTAAGATTTCGAGGCGATGGCATTAAAATTCA

CAG-TAG AAGCCATTTTTTCTTTTCGTTTGCCTTTCCT

CGAAATCTTAGAAGCTC 2463

GAGCTTCTAAGATTTCG 2464 1/1201 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Table 24

Oligonucleotides to produce plants with reduced linolenic acid

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

Reduced linolenic acid [AATAGAACGACAGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGC | 2465 omega-3 fatty acid TCCAATGGCGAGCTAGGTTTTATCAGAATGTGGTTTTAGACCTCTC desaturase CCCAGATTCTACCCTAAACACACAACCTC

Arabidopsis thaliana |GAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAGGTCTAAAACCA | 2466

SerdTerm CATTCTGATAAAACCTAGCTCGCCATTGGAGCCTCTTCCCAAGAAG

TCG-TAG AAAAGAGGAAAAAGTCTCTGTCGTTCTATT

GGCGAGCTIGGTTTTAT 2467 ® ATAAAACCAAGCTCGCC 2468

Reduced linolenic acid | ACGACAGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGCTCCAAT | 2469 omega-3 fatty acid GGCGAGCTCCGTTTGATCAGAATGTGGTTTTAGACCTCTCCCCAG desaturase ATTCTACCCTAAACACACAACCTCTTTTGC

Arabidopsis thaliana |GCAAAAGAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAGGTCTA | 2470

LeuBTerm AAACCACATTCTGATCAAACCGAGCTCGCCATTGGAGCCTCTTCCC

TTA-TGA AAGAAGAAAAGAGGAAAAAGTCTCTGTCGT

CTCGGTTTGATCAGAAT 2471

ATTCTGATCAAACCGAG 2472

Reduced linolenic acid |ACAGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGCTCCAATGGC | 2473 omega-3 fatty acid GAGCTCGGTTTTATGAGAATGTGGTTTTAGACCTCTCCCCAGATTC desaturase TACCCTAAACACACAACCTCTTTTGCCTC

Arabidopsis thaliana |GAGGCAAAAGAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAGGT | 2474

Ser7Term CTAAAACCACATTCTCATAAAACCGAGCTCGCCATTGGAGCCTCTT

Q TCA-TGA CCCAAGAAGAAAAGAGGAAAAAGTCTCTGT

GGTTTTATGAGAATGTG 2475

CACATTCTCATAAAACC 2476

Reduced linolenic acid |AGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGCTCCAATGGCGA | 2477 omega-3 fatty acid GCTCGGTTTTATCATAATGTGGTTTTAGACCTCTCCCCAGATTCTA desaturase CCCTAAACACACAACCTCTTTTGCCTCTA

Arabidopsis thaliana |TAGAGGCAAAAGAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAG | 2478

Glu8Term GTCTAAAACCACATTATGATAAAACCGAGCTCGCCATTGGAGCCTC

GAA-TAA TTCCCAAGAAGAAAAGAGGAAAAAGTCTCT

TTTTATCATAATGTGGT 2479

ACCACATTATGATAAAA

1112/01 01:47 pm 03137.009 — [NY}793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene, B

Plant & Targeted Altering Oligos . Alteration

Reduced linolenic acid A A AGAGCAGAGAGA AAAAGA 48 omega-3 fatty acid TAGCAATGGCGAACTAGGTCTTATCCGAATGTGGCATAAGACCTC desaturase TCCCCAGAATCTACACCACACCCAGATCCAC

Brassica juncea GTGGATCTGGGTGTGGTGTAGATTCTGGGGAGAGGTCTTATGCCA | 2482

LeudTerm CATTCGGATAAGACCTAGTTCGCCATTGCTAGAGCTCTTTTGCTCT

TTG-TAG CTCTCTCTCCCCAGAAGAAGAAGATGATGA

GGCGAACTAGGTCTTAT 2483

ATAAGACGCTAGTTCGCC 2484

Reduced linolenic acid [TCTTCTTCTTCTGGGGAGAGAGAGAGAGCAAAAGAGCTCTAGCAA | 2485 omega-3 fatty acid TGGCGAACTTGGTCTGATCCGAATGTGGCATAAGACCTCTCCCCA ® desaturase GAATCTACACCACACCCAGATCCACTTTCCT

Brassica juncea AGGAAAGTGGATCTGGGTGTGGTGTAGATTCTGGGGAGAGGTCTT | 2486

LeubTerm ATGCCACATTCGGATCAGACCAAGTTCGCCATTGCTAGAGCTCTTT

TTA-TGA TGCTCTCTCTCTCTCCCCAGAAGAAGAAGA

CTTGGTCTGATCCGAAT

ATTCGGATCAGACCAAG

Reduced linolenic acid [TTCTTCTGGGGAGAGAGAGAGAGCAAAAGAGCTCTAGCAATGGCG | 2489 omega-3 fatty acid AACTTGGTCTTATCCTAATGTGGCATAAGACCTCTCCCCAGAATCT desaturase ACACCACACCCAGATCCACTTTCCTCTCCA :

Brassica juncea TGGAGAGGAAAGTGGATCTGGGTGTGGTGTAGATTCTGGGGAGA | 2490

Glu8Term GGTCTTATGCCACATTAGGATAAGACCAAGTTCGCCATTGCTAGA

GAA-TAA GCTCTTTTGCTCTCTCTCTCTCCCCAGAAGAA

TCTTATCCTAATGTGGC 2491

GCCACATTAGGATAAGA 2492 ® Reduced linolenic acid [CTGGGGAGAGAGAGAGAGCAAAAGAGCTCTAGCAATGGCGAACT | 2493 omega-3 fatty acid TGGTCTTATCCGAATGAGGCATAAGACCTCTCCCCAGAATCTACAC desaturase CACACCCAGATCCACTTTCCTCTCCAACACC

Brassica juncea GGTGTTGGAGAGGAAAGTGGATCTGGGTGTGGTGTAGATICTGG | 2494

Cys9Term GGAGAGGTCTTATGCCTCATTCGGATAAGACCAAGTTCGCCATTG

TGT-TGA CTAGAGCTCTTTTGCTCTCTCTCTCTCCCCAG

TCCGAATGAGGCATAAG 2495

CTTATGCCTCATTCGGA 2496 -25 Reduced linolenic acid JATAACAGAR AA i A AR xe omega-3 fatty acid TGGCTGCTGGTTGAGTATTATCAGAATGTGGTTTAAGGCCTCTCCC desaturase AAGAATCTACTCACGACCCAGAATTGGT

Ricinus communis ACCAATTCTGGGTCGTGAGTAGATTCTTGGGAGAGGCCTTAAACC | 2498

Trp5Term ACATTCTGATAATACTCAACCAGCAGCCATTGAAAACCCAGAAGCT

TGG-TGA AAAAATGCAAGAATTCAGCAATTCTGTTAT 14/12/01 01:47 pm ' 03137.009 — JNY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

GCTGGTTGAGTATTATC 2499 : GATAATACTCAACCAGC 2500

Reduced linolenic acid [AGAATTGCTGAATTCTTGCATTTTTAGCTTCTGGGTTTTCAATGGCT | 2501 omega-3 fatty acid GCTGGTTGGGTATGATCAGAATGTGGTTTAAGGCCTCTCCCAAGA desaturase ATCTACTCACGACCCAGAATTGGTTTTAC

Ricinus communis GTAAAACCAATTCTGGGTCGTGAGTAGATTCTTGGGAGAGGCCTT | 2502

Leu7Term AAACCACATTCTGATCATACCCAACCAGCAGCCATTGAAAACCCAG

TTA-TGA AAGCTAAAAATGCAAGAATTCAGCAATTCT

TTGGGTATGATCAGAAT 2503 ® ATTCTGATCATACCCAA 2504

Reduced linolenic acid |ATTGCTGAATTCTTGCATITTTAGCTTCTGGGTTTTCAATGGCTGCT | 2505 omega-3 fatty acid GGTTGGGTATTATGAGAATGTGGTTTAAGGCCTCTCCCAAGAATCT desaturase ACTCACGACCCAGAATTGGTTTTACATC

Ricinus communis GATGTAAAACCAATTCTGGGTCGTGAGTAGATTCTTGGGAGAGGC | 2506

Ser8Term CTTAAACCACATTCTCATAATACCCAACCAGCAGCCATTGAAAACC

TCA-TGA CAGAAGCTAAAAATGCAAGAATTCAGCAAT

GGTATTATGAGAATGTG

CACATTCTCATAATACC 2508

Reduced linolenic acid |TGCTGAATTCTTGCATTTTTAGCTTCTGGGTTTTCAATGGCTGCTG | 2509 omega-3 fatty acid GTTGGGTATTATCATAATGTGGTTTAAGGCCTCTCCCAAGAATCTA desaturase CTCACGACCCAGAATTGGTTTTACATCGA

Ricinus communis TCGATGTAAAACCAATTCTGGGTCGTGAGTAGATTCTTGGGAGAG | 2510

Glu9Term GCCTTAAACCACATTATGATAATACCCAACCAGCAGCCATTGAAAA

GAA-TAA CCCAGAAGCTAAAAATGCAAGAATTCAGCA ® TATTATCATAATGTGGT 2511

ACCACATTATGATAATA 2512

Reduced linolenic acid AA ATCAGAA AGACCA AAGAA omega-3 fatty acid TCTACCCTAAGCCCTGAACTGGGGCAGCCACTTCTGCCTCCTCTC desaturase ACATTAAGTTGAGAATTTCACGTACAGATC

Nicotiana tabacum GATCTGTACGTGAAATTCTCAACTTAATGTGAGAGGAGGCAGAAGT| 2514

Arg22Term GGCTGCCCCAGTTCAGGGCTTAGGGTAGATTCTTGGGAGTGGTCT

AGA-TGA AAGACCACATTCTGATAAAACCCAACTTGC : CTAAGCCCTIGAACTGGG 2515

CCCAGTTCAGGGCTTAG 2516 112001 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos :

Reduced linolenic acid | CTCCCAAGAATCTACCCTAAGCCCAGAACTGGGGCAGCCACTTCT | 2517 . omega-3 fatty acid GCCTCCTCTCACATTTAGTTGAGAATTTCACGTACAGATCTGAGTG desaturase GTTCTGCAATTTCTITGTCTAATACTAATA

Nicotiana tabacum TATTAGTATTAGACAAAGAAATTGCAGAACCACTCAGATCTGTACG | 2518

Lys34Term TGAAATTCTCAACTAAATGTGAGAGGAGGCAGAAGTGGCTGCCCC

AAG-TAG AGTTCTGGGCTTAGGGTAGATTCTTGGGAG

CTCACATTTAGTTGAGA 2519

TCTCAACTAAATGTGAG 2520

Reduced linolenic acid | CAAGAATCTACCCTAAGCCCAGAACTGGGGCAGCCACTTCTGCCT | 2521 omega-3 fatty acid CCTCTCACATTAAGTAGAGAATTTCACGTACAGATCTGAGTGGTTC ® desaturase TGCAATTTCTTTGTCTAATACTAATAAAGA

Nicotiana tabacum TCTTTATTAGTATTAGACAAAGAAATTGCAGAACCACTCAGATCTGT | 2522

Leu35Term ACGTGAAATTCTCTACTTAATGTGAGAGGAGGCAGAAGTGGCTGC

TTG-TAG CCCAGTTCTGGGCTTAGGGTAGATTCTTG

CATTAAGTAGAGAATTT 2523

AAATTCTCTACTTAATG 2524

Reduced linolenic acid |AGAATCTACCCTAAGCCCAGAACTGGGGCAGCCACTTCTGCCTCC | 2525 omega-3 fatty acid TCTCACATTAAGTTGTGAATTTCACGTACAGATCTGAGTGGTTCTG desaturase CAATTTCTTTGTCTAATACTAATAAAGAGA

Nicotiana tabacum TCTCTTTATTAGTATTAGACAAAGAAATTGCAGAACCACTCAGATCT | 2526

Arg36Term GTACGTGAAATTCACAACTTAATGTGAGAGGAGGCAGAAGTGGCT

AGA-TGA GCCCCAGTTCTGGGCTTAGGGTAGATTCT

TTAAGTTGTIGAATTTCA 2527

TGAAATTCACAACTTAA 2528 @® Reduced linolenic acid A ATCAGAA A A A J omega-3 fatty acid GTCTATCCTAAGCCATGAACTGGCCACCCTTTGTTGAATTCCAATC desaturase CCACAAAGCTGAGATTTTCAAGAACAGATC

Sesamum indicum GATCTGTTCTTGAAAATCTCAGCTTTGTGGGATTGGAATTCAACAA | 2530

Arg22Term AGGGTGGCCAGTTCATGGCTTAGGATAGACCCTCGGGAGTGGCC

AGA-TGA TCAGACCACATTCTGATAAAACCCAACTCGC

CTAAGCCATGAACTGGC 2531

GCCAGTTCATGGCTTAG 2532 ‘25 Reduced linolenic acid |CAGAATGTGGTCTGAGGCCACTCCCGAGGGTCTATCCTAAGCCAA | 2533 omega-3 fatty acid GAACTGGCCACCCTTAGTTGAATTCCAATCCCACAAAGCTGAGATT desaturase TTCAAGAACAGATCTTGGAAATGGTTCTTC

Sesamum indicum GAAGAACCATTTCCAAGATCTGTTCTTGAAAATCTCAGCTTTGTGG | 2534

Leu27Term GATTGGAATTCAACTAAGGGTGGCCAGTTCTTGGCTTAGGATAGA

TTG-TAG CCCTCGGGAGTGGCCTCAGACCACATTCTG 1112001 01:47 pm 03137.009 — {NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

CCACCCTTAGTTGAATT 2535

AATTCAACTAAGGGTGG 2536

Reduced linolenic acid |AATGTGGTCTGAGGCCACTCCCGAGGGTCTATCCTAAGCCAAGAA | 2537 omega-3 fatty acid CTGGCCACCCTTTGTAGAATTCCAATCCCACAAAGCTGAGATTTTC desaturase AAGAACAGATCTTGGAAATGGTTCTTCATT

Sesamum indicum AATGAAGAACCATTTCCAAGATCTGTTCTTGAAAATCTCAGCTTTGT[ 2538

Leu28Term GGGATTGGAATTCTACAAAGGGTGGCCAGTTCTTGGCTTAGGATA

TIG-TAG GACCCTCGGGAGTGGCCTCAGACCACATT

CCCTTTGTAGAATTCCA 2539 ® TGGAATTCTACAAAGGG

Reduced linolenic acid |CTCCCGAGGGTCTATCCTAAGCCAAGAACTGGCCACCCTTIGTTG | 2541 omega-3 fatty acid AATTCCAATCCCACATAGCTGAGATTTTCAAGAACAGATCTTGGAA desaturase ATGGTTCTTCATTCTGTTTGTCGAGTGGGA

Sesamum indicum TCCCACTCGACAAACAGAATGAAGAACCATTTCCAAGATCTGTTCT | 2542

Lys34Term TGAAAATCTCAGCTATGTGGGATTGGAATTCAACAAAGGGTGGCC

AAG-TAG AGTTCTTGGCTTAGGATAGACCCTCGGGAG

ATCCCACATAGCTGAGA

TCTCAGCTATGTGGGAT

Reduced linolenic acid A AGA A | A AA A AAGAA A 4 omega-3 fatty acid GAAGTCTATGAGTTAGGTCGTCAGAGAGCTAGCCATCGTGTTCGC desaturase ACTAGCTGCTGGAGCTGCTTACCTCAACAAT

Brassica napus ATTGTTGAGGTAAGCAGCTCCAGCAGCTAGTGCGAACACGATGGC | 2546

Tyr3Term TAGCTCTCTGACGACCTAACTCATAGACTTCCATGGATTCTTAACC

TAC-TAG CAGCAATGCTTAGGTATCGCCGCTCTGATG ® ATGAGTTAGGTCGTCAG 2547

CTGACGACCTAACTCAT 2548

Reduced linolenic acid |GCGGCGATACCTAAGCATTGCTGGGTTAAGAATCCATGGAAGTCT | 2549 omega-3 fatty acid ATGAGTTACGTCGTCTGAGAGCTAGCCATCGTGTTCGCACTAGCT desaturase GCTGGAGCTGCTTACCTCAACAATTGGCTTG

Brassica napus CAAGCCAATTGTTGAGGTAAGCAGCTCCAGCAGCTAGTGCGAACA | 2550

Arg6Term CGATGGCTAGCTCTCAGACGACGTAACTCATAGACTTCCATGGATT

AGA-TGA CTTAACCCAGCAATGCTTAGGTATCGCCGC ‘ ACGTCGTCTGAGAGCTA 2551

TAGCTCTCAGACGACGT 2552 111201 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos : Alteration

Reduced linolenic acid | GCGATACCTAAGCATTGCTGGGTTAAGAATCCATGGAAGTCTATGA| 2553 omega-3 fatty acid GTTACGTCGTCAGATAGCTAGCCATCGTGTTCGCACTAGCTGCTG desaturase GAGCTGCTTACCTCAACAATTGGCTTGTTT

Brassica napus AAACAAGCCAATTGTTGAGGTAAGCAGCTCCAGCAGCTAGTGCGA | 2554

Glu7Term ACACGATGGCTAGCTATCTGACGACGTAACTCATAGACTTCCATG

GAG-TAG GATTCTTAACCCAGCAATGCTTAGGTATCGC

TCGTCAGATAGCTAGCC 2555

GGCTAGCTATCTGACGA omega-3 fatty acid TTCGCACTAGCTGCTIGAGCTGCTTACCTCAACAATTGGCTTGTTT @® desaturase GGCCTCTCTATTGGATTGCTCAAGGAACCA

Brassica napus TGGTTCCTTGAGCAATCCAATAGAGAGGCCAAACAAGCCAATTGTT | 2558

Gly17Term GAGGTAAGCAGCTCAAGCAGCTAGTGCGAACACGATGGCTAGCT

GGA-TGA CTCTGACGACGTAACTCATAGACTTCCATGG

TAGCTGCTTGAGCTGCT 2559

AGCAGCTCAAGCAGCTA 2560

Reduced linolenic acid AA ATCAGAA AGA ACTACCAAGAA B omega-3 fatty acid TATACCCAAAGCCCTGAATAGGGTCTTCTTCCGTTTGCGCCACCAA desaturase TTTAAATCTGAGAAGAATTTCACCTTCAC

Solanum tuberosum |GTGAAGGTGAAATTCTTCTCAGATTTAAATTGGTGGCGCAAACGGA| 2562

Arg22Term AGAAGACCCTATTCAGGGCTTTGGGTATATTCTTGGTAGTGGTCTA

AGA-TGA AGACCACATTCTGATAGAACCCAACTTGC

CAAAGCCCTGAATAGGG 2563

CCCTATTCAGGGCTTTG

® omega-3 fatty acid GTCTTCTTCCGTTTGAGCCACCAATTTAAATCTGAGAAGAATTTCA desaturase CCTTCACCTATACGAACAGATCGGAATTGT

Solanum tuberosum |ACAATTCCGATCTGTTCGTATAGGTGAAGGTGAAATTCTTCTCAGA | 2566

Cys29Term TTTAAATTGGTGGCTCAAACGGAAGAAGACCCTATTCTGGGCTTTG

TGC-TGA GGTATATTCTTGGTAGTGGTCTAAGACCA

TCCGTTTGAGCCACCAA 2567

TTGGTGGCTCAAACGGA 2568

Reduced linolenic acid | CACTACCAAGAATATACCCAAAGCCCAGAATAGGGTCTTCTTCCGT | 2569 omega-3 fatty acid TTGCGCCACCAATTGAAATCTGAGAAGAATTTCACCTTCACCTATA desaturase CGAACAGATCGGAATTGTTGGGCATTGAG

Solanum tuberosum CTCAATGCCCAACAATTCCGATCTGTTCGTATAGGTGAAGGTGAAA 2570

Leu33Term TTCTTCTCAGATTTCAATTGGTGGCGCAAACGGAAGAAGACCCTAT

TTA-TGA TCTGGGCTTTGGGTATATTCTTGGTAGTG 1112/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos ) Alteration

CACCAATTGAAATCTGA 2571

TCAGATTTCAATTGGTG 2572

Reduced linolenic acid AGAATATACCCAAAGCCCAGAATAGGGTCTTCTTCCGTTTGCGCCA| 2573 omega-3 fatty acid CCAATTTAAATCTGTGAAGAATTTCACCTTCACCTATACGAACAGAT desaturase CGGAATTGTTGGGCATTGAGGGTAAGTG

Solanum tuberosum {CACTTACCCTCAATGCCCAACAATTCCGATCTGTTCGTATAGGTGA | 2574

Arg36Term AGGTGAAATTCTTCACAGATTTAAATTGGTGGCGCAAACGGAAGAA

AGA-TGA GACCCTATTCTGGGCTTTGGGTATATTCT

TAAATCTGIGAAGAATT 2575 [ AATTCTTCACAGATTTA 2576

Reduced linolenic acid ATTA AGT i A omega-3 fatty acid TATGGCAAGTTGAGTGATTTCAGAATGTGGGCTAAGGCCACTTCC desaturase AAGAATCTATGCCAGGCCCAGAAGTGGA

Petroselinum crispum |TCCACTTCTGGGCCTGGCATAGATTCTTGGAAGTGGCCTTAGCCC | 2578

Trp4Tem ACATTCTGAAATCACTCAACTTGCCATAGGTGACTCAGAACTCAAA

TGG-TGA AAAAACAAAGAAGAGGAGGATAATAAAGAG

GCAAGTTGAGTGATTTC

GAAATCACTICAACTTGC 2580 omega-3 fatty acid AGTTGGGTGATTTGAGAATGTGGGCTAAGGCCACTTCCAAGAATC desaturase TATGCCAGGCCCAGAAGTGGAGCTTCATG

Petroselinum crispum |CATGAAGCTCCACTTCTGGGCCTGGCATAGATTCTTGGAAGTGGC | 2582

Ser7Term CTTAGCCCACATTCTCAAATCACCCAACTTGCCATAGGTGACTCAG

TCA-TGA AACTCAAAAAAAACAAAGAAGAGGAGGATA @ GGTGATTTGAGAATGTG | 2583

CACATTCTCAAATCACC

Reduced linolenic acid |[TCCTCCTCTTCTTTGTTTTTTTTGAGTTCTGAGTCACCTATGGCAAG | 2585 omega-3 fatty acid TTGGGTGATTTCATAATGTGGGCTAAGGCCACTTCCAAGAATCTAT desaturase GCCAGGCCCAGAAGTGGAGCTTCATGTT

Petroselinum crispum |AACATGAAGCTCCACTTCTGGGCCTGGCATAGATTCTTGGAAGTG | 2586

Glu8Term GCCTTAGCCCACATTATGAAATCACCCAACTTGCCATAGGTGACTC

GAA-TAA AGAACTCAAAAAAAACAAAGAAGAGGAGGA : TGATTTCATAATGTGGG 2587

CCCACATTATGAAATCA 2588 11/42/01 01:47 pm 03137.009 — [NY}793559.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Reduced linolenic acid |CTCTTCTTTGTTTTTTTITGAGTTCTGAGTCACCTATGGCAAGTTGGG | 2589 : omega-3 fatty acid TGATTTCAGAATGAGGGCTAAGGCCACTTCCAAGAATCTATGCCA desaturase GGCCCAGAAGTGGAGCTTCATGTTTCAAC

Petroselinum crispum {GTTGAAACATGAAGCTCCACTTCTGGGCCTGGCATAGATTCTTGG | 2590

Cys9Term AAGTGGCCTTAGCCCTCATTCTGAAATCACCCAACTTGCCATAGGT

TGT-TGA GACTCAGAACTCAAAAAAAACAAAGAAGAG

TCAGAATGAGGGCTAAG 2591

CTTAGCCCICATTCTGA 2592

Reduced finolenic acid JATGAAGCAGCAACAGTACAAAGACA AA AAJ A 0 omega-3 fatty acid ATGGTTTTCATGCTTAAGAAGAAGAAGAAGAAGAGGATTTCGACTT ® desaturase AAGCAATCCTCCTCCATTCAATATTGGTC

Vernicia fordii GACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATCCTCTTC | 2594

Lys21Term TTCTTCTTCTTCTTAAGCATGAAAACCATTAACGCCATTTAGAATTG

AAA-TAA CGGTGTCTTTGTACTGTTGCTGCTTCAT

TTCATGCTTAAGAAGAA 2595

TTCTTCTTAAGCATGAA 2596

Reduced linolenic acid [AAGCAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTAATG | 2597 omega-3 fatty acid GTTTTCATGCTAAATAAGAAGAAGAAGAAGAGGATTTCGACTTAAG desaturase CAATCCTCCTCCATTCAATATTGGTCAGA

Vernicia fordii TCTGACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATCCTC | 2598

Glu22Term TTCTTCTTCTTCTTATTTAGCATGAAAACCATTAACGCCATTTAGAA

GAA-TAA TTGGGGTGTCTTTGTACTGTTGCTGCTT

ATGCTAAATAAGAAGAA 2599

TTCTTCTTATTTAGCAT 2600 ® Reduced linolenic acid | CAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTAATGGTT | 2601 . 20 omega-3 fatty acid TTCATGCTAAAGAATAAGAAGAAGAAGAGGATTTCGACTTAAGCAA desaturase TCCTCCTCCATTCAATATTGGTCAGATCC

Vernicia fordii GGATCTGACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATC | 2602

Glu23Term CTCTTCTTCTTCTTATTCTTTAGCATGAAAACCATTAACGCCATTTA

GAA-TAA GAATTGGGGTGTCTTTGTACTGTTGCTG

CTAAAGAATAAGAAGAA 2603

TTCTTCTTATICTTTAG 2604

Reduced linolenic acid |CAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTAATGGTT | 2605 omega-3 fatty acid TTCATGCTAAAGAATAAGAAGAAGAAGAGGATTTCGACTTAAGCAA desaturase TCCTCCTCCATTCAATATTGGTCAGATCC

Vernicia fordii GGATCTGACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATC| 2606

Glu24Term CTCTTCTTCTTCTTATTCTTTAGCATGAAAACCATTAACGCCATTTA

GAA-TAA GAATTGGGGTGTCTITGTACTGTTGCTG 11/12/01 01:47 pm 03137.009 — [NY}793558.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos

Alteration

CTAAAGAATAAGAAGAA 2607 : TTCTTCTTATTCTTTAG 2608 reduced linolenic acid AAGCACA ACAACA AA A AA HUY omega-3 fatty acid AGAAGATCAAGCTTAGTTTGATCCAAGTGCTCCACCACCCTTCAAG desaturase ATTGCAAATATCAGAGCAGCAATTCCAAAA

Glycine max TTTTGGAATTGCTGCTCTGATATTTGCAATCTTGAAGGGTGGTGGA | 2610

Tyr21Term GCACTTGGATCAAACTAAGCTTGATCTTCTTTCCCTGCACCATTAC

TAT-TAG CAACATGTTGTAGAGGCTGTGCTTGGACC

CAAGCTTAGTTTGATCC 2611 ® GGATCAAACTAAGCTTG 2612

Reduced Inokenc acd |GGTAATGGTGCAGGGAAAGARGATCARGCT TATTTTGATCOAAGT | 2613 omega-3 fatty acid GCTCCACCACCCTTCTAGATTGCAAATATCAGAGCAGCAATTCCAA desaturase AACATTGCTGGGAGAAGAACACATTGAGAT

Glycine max ATCTCAATGTGTTCTTCTCCCAGCAATGTTTTGGAATTGCTGCTCT 2614

Lys31Term GATATTTGCAATCTAGAAGGGTGGTGGAGCACTTGGATCAAAATAA

AAG-TAG GCTTGATCTTCTTTCCCTGCACCATTACC

CACCCTTCTAGATTGCA 2615

TGCAATCTAGAAGGGTG 2616

Reduced linolenic acid |AAAGAAGATCAAGCTTATTTTGATCCAAGTGCTCCACCACCCTTCA | 2617 omega-3 fatty acid AGATTGCAAATATCTGAGCAGCAATTCCAAAACATTGCTGGGAGAA desaturase GAACACATTGAGATCTCTGAGTTATGTTC

Glycine max GAACATAACTCAGAGATCTCAATGTGTTCTTCTCCCAGCAATGTTTT| 2618

Arg36Term GGAATTGCTGCTCAGATATTTGCAATCTTGAAGGGTGGTGGAGCA

AGA-TGA CTTGGATCAAAATAAGCTTGATCTTCTTT

Qo CAAATATCTGAGCAGCA 2619

TGCTGCTCAGATATTTG 2620 omega-3 fatty acid GAGCAGCAATTCCATAACATTGCTGGGAGAAGAACACATTGAGAT desaturase CTCTGAGTTATGTTCTGAGGGATGTGTTGG

Glycine max CCAACACATCCCTCAGAACATAACTCAGAGATCTCAATGTGTTCTT | 2622

LeudtTerm CTCCCAGCAATGTTATGGAATTGCTGCTCTGATATTTGCAATCTTG

AAA-TAA AAGGGTGGTGGAGCACTTGGATCAAAATA

CAATTCCATAACATTGC 2623

GCAATGTTATGGAATTG 2624 111201 01:47 pm 03137.009 — [NY]793550.1

AMENDED SHEET

28-12-2001 01939797

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration ey} NO:

Reduced Tinolenic acid [CA A i A A y: 3 omega-3 fatty acid CCGGCTCGTGCTCTCCTAGTGCTCGGGCCTCGCGCCCGTCCGCE desaturase GCCTGCGCGCCGGCCGGGGCBCCATTGCGGCGC

Zea mays GCGCCGCAATGGCGCCCCGGCCGGCGCGCAGGCGGCGGACGG | 2626

Glu8Term GCGCGAGGCCCGAGCACTAGGAGAGCACGAGCCGGBCCATTGC

GAG-TAG CGCCGTCAGCGGGGCGGGTGCGGGTGCGGGTGGATG

TeCeTCeneTeCis aw

CGAGCACTAGGAGAGCA 2628

Reduced linolenic acid | ACCCGCACCCGCACCCGCCCCGCTGACGGCGGCAATGGCCCGG | 2629 omega-3 fatty acid CTCGTGCTCTCCGAGTGATCGGGCCTCGCGCCCGTCCGCCGCCT ® desaturase GCGCGCCGGCCGGGGCGCCATTGCGGCGCGGTCA

Zea mays TGACCGCGCCGCAATGGCGCCCCGGCCGGCGCGCAGGCGGCGG | 2630

Cys9Term ACGGGCGCGAGGCCCGATCACTCGGAGAGCACGAGCCGGGCCA

TGC-TGA TTGCCGCCGTCAGCGGGGCGGGTGCGGGRTGCGGET

I CC I

AGGCCCGATCACTCGGA 2632

Reduced linolenic acid | CCGCACCCGCACCCGCCCCGCTGACGGCGGCAATGGCCCGGCT | 2633 omega-3 fatty acid CGTGCTCTCCGAGTGCTAGGGCCTCGCGCCCGTCCGCCGCCTGC desaturase GCGCCGGCCGGGGCGCCATTGCGGCGCGGTCACC

Zea mays GGTGACCGCGCCGCAATGGCGCCCCGGCCGGCGCGCAGGCGGE | 2634

Ser10Term GGACGGGCGCGAGGCCCTAGCACTCGGAGAGCACGAGCCGGGC

TCG-TAG CATTGCCGCCGTCAGCGGGGCGGRTGCGGGTGCGE

CGAGTGCTAGGGCCTCG 2635

CGAGGCCCTAGCACTCG 2636 ® Reduced linolenic acid {GCTCGGGCCTCGCGCCCGTCCGCCGCCTGCGCGCCGGCCGGRG | 2637 omega-3 fatty acid CGCCATTGCGGCGCGGTBGACCCCCCGCGCTCTCCGCGGCGCCE desaturase CGCCGTCGTCCCGCGTCCGCGTCCATCCACCGCGA

Zea mays TCGCGGTGGATGGACGCGGACGCGGGACGACGGCGCGGCGCCE | 2638

Ser29Term CGGAGAGCGCGGGGGGTCACCGCGCCGCAATGGCGCCCCGGLC i TCA-TGA GGCGCGCAGGCGGCGGACGGGCGCGCAGGCCCGAGC a

Reduced linolenic acid A ACA ACAGA A A of omega-3 fatty acid CCCCGCAATGAGGCCGTAGCAGGAGGCGAGCTGCAAGGCCACCG desaturase AGGACCACCGCTCCGAGTTCGACGCCGCCAAGC ) Triticum aestivum GCTTGGCGGCGTCGAACTCGGAGCGGTGGTCCTCGGTGGCCTTG | 2642

GluBTerm CAGCTCGCCTCCTGCTACGGCCTCATTGCGGGGGCCATGGCCGC

GAG-TAG GGATGGATCTGTGCGTGTGCGTGGGCGAGGGEE 11201 01:47 pm 03137.009 — [NY]783559.1

AMENDED SHEET

28-12-2001 01939797

B Phenotype, Gene,

Plant & Targeted Altering Oligos i Alteration

TM TITTITITIIT ROORR BERiaytiyez >

TGAGGCCGTAGCAGGAG 2643

CTOCTGCTACGGCCTCA | %#

Reduced linolenic acid |CCTCCCCCACGCACACGCACAGATCCATCCGCGGCCATGGCCCC | 2845 omega-3 fatty acid CGCAATGAGGCCGGAGTAGGAGGCGAGCTGCAAGGCCACCGAG desaturase GACCACCGCTCCGAGTTCGACGCCGCCAAGCCEGC

Triticum aestivum GCGGCTTGGCGGCGTCGAACTCGGAGCGGTGGTCCTCGGTGRCC| 2646

GIn9Term TTGCAGCTCGCCTCCTACTCCGGCCTCATTGCGGGGGCCATGGC

CAG-TAG CGCGGATGGATCTGTGCGTGTGCGTGGGGGAGG

GGCCGGAGTAGGAGGCG 2647 ® CooCTCCTACTCCGGC. | mm

Reduced linolenic acid {CCCCCACGCACACGCACAGATCCATCCGCGGCCATGGCCCCCGE [| 2649 omega-3 fatty acid AATGAGGCCGGAGCAGTAGGCGAGCTGCAAGGCCACCGAGGACC desaturase ACCGCTCCGAGTTCGACGCCGCCAAGCCGCCGC

Triticum aestivum GCGGCGGCTTGGCGGCGTCGAACTCGGAGCGGTGGTCCTCGGT | 2650

Glu10Term GGCCTTGCAGCTCGCCTACTGCTCCGGCCTCATTGCGGGGGCCA

GAG-TAG TGGCCGCCGCGATGGATCTGTGCGTGTGCGTGGGGE

CGGAGCAGTAGGCGAGC 2651

OCTCGCCTACTGCTCCG [26%

Reduced linolenic acid JACGCACAGATCCATCCGCGGCCATGGCCCCCGCAATGAGGCCGG | 2653 omega-3 fatty acid AGCAGGAGGCGAGCTGAAAGGCCACCGAGGACCACCGCTCCGA desaturase GTTCGACGCCGCCAAGCCGCCGCCCTTCCGCATC

Triticum aestivum GATGCGGAAGGGCGGCGGCTTGGCGGCGTCGAACTCGGAGCGG | 2654

Cys13Term TGGTCCTCGGTGGCCTTTICAGCTCGCCTCCTGCTCCGGCCTCATT

TGC-TGA GCGGGGGCCATGGCCGCGGATGGATCTGTGCGT ® GCGAGCTGAAAGGCCAC 2655

GTGGCCTTICAGCTCGC 2656

Reduced linolenic acid ACAAATCACAAE AATCAGA ACCACGACA 0 omega-3 fatty acid CGGCAATGGCGGCGTAGGCGACCCAGGAGGCCGACTGCAAGGE desaturase TTCCGAGGACGCCCGTCTCTTCTTCGACGCCGC

Oryza sativa GCGGCGTCGAAGAAGAGACGGGCGTCCTCGGAAGCCTTGCAGTC | 2658

SerdTerm GGCCTCCTGGGTCGCCTACGCCGCCATTGCCGCCRGGGTGTCGT

TCG-TAG GGTGGATCTGATTCCGATTTGTGATTTGTGAAG

GaGTeeeCTAGSEeeCe | 7m ee _] SYYY 1412/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

28-12-2001 01939797 —

Phenotype, Gene,

Plant & Targeted Altering Oligos . Alteration

TI TN EE RTC VT NC TT CY ee

Reduced linolenic acid ATCACAAATCGGAATCAGATCCACCACGACACCCCGGCGGCAATG | 2661 , omega-3 fatty acid GCGGCGTCGGCGACCTAGGAGGCCGACTGCAAGGCTTCCGAGGA desaturase CGCCCGTCTCTTCTTCGACGCCGCCAAGCCCC

Oryza sativa GGGGCTTGGCGGCGTCGAAGAAGAGACGGGCGTCCTCGGAAGC | 2662

GIn7Term CTTGCAGTCGGCCTCCTAGGTCGCCGACGCCGCCATTGCCGCCS

CAG-TAG GGGTGTCGTGGTGGATCTGATTCCGATTTGTGAT

CGGCGACCTAGGAGGCC 2663

GGCCTCCTAGGTCGCC6 ~~ — | 2664

Reduced linolenic acid |ACAAATCGGAATCAGATCCACCACGACACCCCGGCGGCAATGGC 2665 omega-3 fatty acid GGCGTCGGCGACCCAGTAGGCCGACTGCAAGGCTTCCGAGGACG @ desaturase CCCGTCTCTTCTTCGACGCCGCCAAGCCCCCGC

Oryza sativa GCGGGGGCTTGGCGGCGTCGAAGAAGAGACGGGCGTCCTCGGA | 2666

Glu8Term AGCCTTGCAGTCGGCCTACTGGGTCGCCGACGCCGCCATTGCCG

GAG-TAG CCGGGGTGTCGTGATGGATCTGATTCCGATTTGT

CGACCCAGTAGGCCGAC 2667 omega-3 fatty acid CCCAGGAGGCCGACTGAAAGGCTTCCGAGGACGCCCGTCTCTTC desaturase TTCGACGCCGCCAAGCCCCCGCCCTTCCGCATC

Oryza sativa GATGCGGAAGGGCGGGGGCTTGGCGGCGTCGAAGAAGAGACGG | 2670

Cys10Term GCGTCCTCGGAAGCCTTICAGTCGGCCTCCTGGGTCGCCGACGE

TGC-TGA CGCCATTGCCGCCGGGGTGTCGTGGTGGATCTGA

GCCGACTGAAAGGCTTC 2671

GAAGCCTTTICAGTCGGC 2672 11/42/01 01:47 pm 03137.009 — [NY]793559.1

AMENDED SHEET

Claims (74)

204-3 WHAT IS CLAIMED IS:

1. A method of targeted alteration of a plant chromosome sequence, the method comprising: introducing a sequence-altering oligonucleotide into a plant cell in vitro, wherein said sequence-altering oligonucleotide: is a single-stranded nonhairpin oligonucleotide 17 - 121 nucleotides in length; has an unmodified DNA domain of at least 8 contiguous deoxyribonucleotides; is fully complementary in sequence to a first strand of the cell's chromosomal DNA at a chromosomal target sequence, except for one or two mismatches positioned (i) within said oligonucleotide's unmodified DNA domain and (ii) at least 8 nucleotides from said oligonucleotide's 5' and 3' termini; and has at least one terminal chemical modification, wherein said sequence alteration confers a phenotype selected from the group consisting of: imidazolinone herbicide resistance, sufonylurea herbicide resistance, porphyric herbicide resistance, triazine resistance, male sterility, abiotic stress tolerance, albinism, altered amino acid content, altered starch metabolism, and altered fatty acid content.

2. A method of targeted alteration of a plant chromosome sequence, the method comprising: introducing a sequence-altering oligonucleotide into a plant cell in vitro, wherein said sequence-altering oligonucleotide: } is a single-stranded nonhairpin oligonucleotide 17 - 121 nucleotides in length; has an unmodified DNA domain of at least 8 contiguous deoxyribonucleotides; is fully complementary in sequence to a first strand of the cell's chromosomal DNA at a chromosomal target sequence, except for one or two mismatches Amended Sheet —- 2004-08-20

-203- C positioned (i) within said oligonucleotide's unmodified DNA domain and (ii) at least 8 nucleotides from said oligonucleotide’s §' and 3' termini; and has at least one terminal chemical modification, wherein said plant cell is selected from the group of plant species consisting of: Chlamydomonas reinhardtii, Physcomitrella patens, cauliflower, artichoke, apples, mangoes, berries, kiwifruit, grapes, bell peppers, cherries, cucumber, melons, nuts, walnuts, peanut, orange, peach, pear, plum, strawberry, tomato, alfalfa, cabbage, endive, leek, lettuce arrowroot, beet, carrot, cassava, turnip, radish, yam, sweet potato, oilseeds, beans, pea, cowpea, mothbean, sorghum, barley, rice, millet, sunflower, oats, chickpea, tubers, kohirabi, potato, flax, cotton, pine, oak, eucalyptus, turfgrass, petunia, hyacinth, carnation, delphinium, Job's tears, snapdragon, poppy, lilac, hydrangea, roses, Gallica roses, Alba roses, Damask roses, Damask Perpetual roses, Centifolia roses, China roses, Tea roses, Hybrid Tea roses, and ornamental goldenrods.

3. A method of targeted alteration of a plant chromosome sequence, the method comprising: introducing a sequence-altering oligonucleotide into a plant cell in vitro, wherein said sequence-altering oligonucleotide: is a single-stranded nonhairpin oligonucleotide 17 - 121 nucleotides in length; has an unmodified DNA domain of at least 8 contiguous deoxyribonuclectides; is fully complementary in sequence to a first strand of the cell's chromosomal DNA at a chromosomal target sequence, except for one or two mismatches positioned (i) within said oligonucleotide's unmodified DNA domain and (ii) at least 8 nucleotides from said oligonucleotide's 5' and 3' termini; and has at least one terminal chemical modification, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 1 —- 2672.

4. The method of any one of claims 1 - 3, wherein said at least one terminal modification is selected from the group consisting of: at least one terminal Amended Sheet - 2004-08-20

208.5 phosphorothioate linkage, at least one terminal locked nucleic acid (LNA) residue, and at least one 2'-O-Me residue.

5. The method of claim 4, wherein said at least one terminal modification is at least one terminal phosphorothioate linkage.

6. The method of claim 5, wherein said at least one terminal modification is a plurality of contiguous terminal phosphorothioate linkages.

7. The method of claim 6, wherein said at least one terminal modification is at least three contiguous terminal phosphorothioate linkages.

8. The method of any one of claims 1 — 7, wherein both of said oligonucleotide's termini have terminal modifications.

9. The method of claim 8, wherein both of said oligonucleotide’s termini have at least one terminal phosphorothioate linkage.

10. The method of claim 9, wherein both of said oligonucleotide's termini have a plurality of contiguous phosphorothioate linkages.

11. The method of claim 10, wherein both of said oligonucleotide's termini have three contiguous terminal phosphorothioate linkages.

12. The method of any one of claims 1 - 11, wherein said sequence alteration is a substitution of at least one base.

13. The method of any one of claims 1 — 11, wherein said sequence alteration is a deletion of at least one base.

14. The method of any one of claims 1 ~ 11, wherein said alteration is an insertion of at least one base. Amended Sheet - 2004-08-20

207k

15. The method of any one of claims 1 - 14, wherein said oligonucleotide is at least 25 nucleotides in length.

16. The method of any one of claims 1 — 15, wherein said oligonucleotide is no more than 74 nucleotides in length.

17. The method of any one of claims 1 — 16, wherein said first strand is the nontranscribed strand of the chromosomal target.

18. The method of any one of claims 1 — 17, wherein the sequences of said oligonucleotide deoxyribonucleotide domain and of the target nucleic acid first strand are mismatched at a single nucleotide.

19. The method of any one of claims 1 - 17, wherein the sequences of said oligonucleotides deoxyribonucleotide domain and of its complement on the target nucleic acid first strand are mismatched at two or more nucleotides.

20. The method of any one of claims 2 — 19, wherein said sequence alteration confers a phenotype of glyphosate resistance.

21. The method of claim 20, wherein said oligonucleotide comprises a sequence selected from the group of SEQ ID NOs: 1 - 36.

22. The method of any one of claims 2 - 19, wherein said sequence alteration confers a phenotype of herbicide resistance.

23. The method of any one of claims 1 — 19, wherein said sequence alteration confers a phenotype of resistance to an imidazolinone herbicide.

24. The method of claim 23, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 49 - 56, 69 - 72, 81-88, 97 Amended Sheet — 2004-08-20

-20F7 -100, 109 - 112, 125-128, 141 — 144, 164 — 168, 181 — 184, 193 - 196, 209 - 212, 221 - 224, 237 - 240, 253 - 256, 265 - 268, 281 ~ 284, and 297 - 300.

25. The method of claim 22, wherein said herbicide is a sufonylurea herbicide.

26. The method of claim 25, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 27 — 48, 57 - 68, 73 ~ 80, 89 -96, 101 - 108, 113 - 124, 129 - 140, 145 — 152, 157 - 163, 169 — 180, 185-192, 197 - 208, 213 - 220, 225 - 236, 241 - 252, 257 - 264, 269 - 280, and 285 — 296.

27. The method of claim 22, wherein said herbicide is a porphyric herbicide.

28. The method of claim 27, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 301 — 328.

29. The method of claim 22, wherein said herbicide is a triazine herbicide.

30. The method of claim 29, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 329 - 460.

31. The method of any one of claims 1 - 19, wherein said sequence alteration confers a phenotype of male sterility.

32. The method of claim 31, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 461 - 892.

33. The method of any one of claims 1 - 19, wherein said sequence alteration confers a phenotype of stress tolerance. Amended Sheet — 2004-08-20

34. The method of claim 33, wherein said sequence alteration confers salt tolerance.

35. The method of claim 34, wherein said oligonucleotide comprises a sequence selected from the group of SEQ ID NOs: 893 — 988.

36. The method of claim 33, wherein said sequence alteration confers tolerance to freezing.

37. The method of claim 36, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 989 — 1008.

38. The method of claim 33, wherein said sequence alteration confers lead tolerance.

39. The method of claim 38, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 1009 — 1068.

40. The method of any one of claims 1 - 19, wherein said sequence alteration confers a phenotype of resistance to 2,4-DB.

41. The method of claim 40, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 1069 — 1188.

42. The method of any one of claims 1 — 19, wherein said sequence alteration confers an albino phenotype.

43. The method of claim 42, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 1189 - 1288. Amended Sheet — 2004-08-20 wl >a

44, The method of any one of claims 1 - 19, wherein said sequence alteration confers a phenotype of altered amino acid content.

45. The method of claim 44, wherein said phenotype is methionine overproduction.

46. The method of claim 45, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 1289 - 1392.

47. The method of claim 44, wherein said phenotype is lysine overproduction.

48. The method of claim 47, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 1393 - 1488.

49. The method of claim 44, wherein said phenotype is tryptophan overproduction.

50. The method of claim 49, wherein said oligonucleotide has a sequence selected from the group consisting of SEQ ID NOs: 1489 - 1508.

51. The method of any one of claims 1 — 19, wherein said sequence alteration confers a phenotype of altered starch metabolism.

52. The method of claim 51, wherein said phenotype is increased starch production.

53. The method of claim §2, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 1509 - 1704.

54. The method of claim 51, wherein said phenotype is production of waxy starch. Amended Sheet — 2004-08-20

55. The method of claim 54, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 1705 - 2064.

56. The method of any one of claims 1 - 19, wherein said sequence alteration confers a phenotype of altered fatty acid content.

57. The method of claim 56, wherein said sequence alteration causes reduced levels of palmitate.

58. The method of claim 57, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 2065 - 2224.

59. The method of claim 56, wherein said sequence alteration causes increased levels of stearate.

60. The method of claim 53, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 2225 - 2464.

61. The method of claim 56, wherein said sequence alteration causes reduced levels of linolenic acid.

62. The method of claim 61, wherein said oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NOs: 2465 - 2672.

63. The method of any one of claims 1 - 62, wherein said oligonucleotide is introduced into said cells by biolistic or particle bombardment.

64. The method of any one of claims 1 — 62, wherein said oligonucleotide is introduced into said cells by electroporation, using polycations, using cationic using lipids, using liposomes, using polyethylenimine (PE!), or by microinjection. Amended Sheet — 2004-08-20 w=

65. The method of claim 1 or claim 3, wherein said plant cell is selected from the group of plant species consisting of: canola, Arabidopsis, tobacco, banana, maize, soybean, wheat, spinach, Chlamydomonas reinhardtii, Physcomitrella patens, cauliflower, artichoke, apples, mangoes, berries, kiwifruit, grapes, bell peppers, cherries, cucumber, melons, nuts, walnuts, peanut, orange, peach, pear, plum, strawberry, tomato, alfalfa, cabbage, endive, leek, lettuce arrowroot, beet, carrot, cassava, turnip, radish, yam, sweet potato, oilseeds, beans, pea, cowpea, mothbean, sorghum, barley, rice, millet, sunflower, oats, chickpea, tubers, kohlrabi, potato, flax, cotton, pine, oak, eucalyptus, turfgrass, petunia, hyacinth, carnation, delphinium, Job's tears, snapdragon, poppy, lilac, hydrangea, roses, Gallica roses, Alba roses, Damask roses, Damask Perpetual roses, Centifolia roses, China roses, Tea roses, Hybrid Tea roses, and ornamental goldenrods.

66. The method of claim 65, wherein said plant cell is selected from the group of plant species consisting of: canola, maize, soybean, wheat, and rice.

67. The method of claim 66, wherein said plant cell is a canola cell.

68. The method of claim 66, wherein said plant cell is a maize cell.

69. The method of claim 66, wherein said plant cell is a soybean cell.

70. The method of claim 66, wherein said plant cell is a wheat cell.

71. The method of claim 66, wherein said plant cell is a rice cell.

72. The method of any one of claims 1 — 71, further comprising the later step of: generating a plant from said cell.

73. Aplant having an altered chromosomal sequence produced by the method of claim 72. Amended Sheet —- 2004-08-20

74. A seed derived from a plant according to claim 73. Amended Sheet — 2004-08-20