EP1068333A1 - Homologues de la proteine r1 de phosphorylation de l'amidon - Google Patents

Homologues de la proteine r1 de phosphorylation de l'amidon

Info

Publication number
EP1068333A1
EP1068333A1 EP99916471A EP99916471A EP1068333A1 EP 1068333 A1 EP1068333 A1 EP 1068333A1 EP 99916471 A EP99916471 A EP 99916471A EP 99916471 A EP99916471 A EP 99916471A EP 1068333 A1 EP1068333 A1 EP 1068333A1
Authority
EP
European Patent Office
Prior art keywords
starch
nucleic acid
protein
encoding
phosphorylation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP99916471A
Other languages
German (de)
English (en)
Inventor
Robert F. Cressman
Stephen M. Allen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
EIDP Inc
Original Assignee
EI Du Pont de Nemours and Co
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by EI Du Pont de Nemours and Co filed Critical EI Du Pont de Nemours and Co
Publication of EP1068333A1 publication Critical patent/EP1068333A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • C12N15/8245—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis

Definitions

  • This invention is in the field of plant molecular biology. More specifically, this invention pertains to nucleic acid fragments encoding starch Rl phosphorylation proteins in plants and seeds.
  • Amylose is an unbranched chain of up to several thousand ⁇ -D-glucopyranose units linked by ⁇ -1,4 glycosidic bonds.
  • Amylopectin is a highly branched molecule made up of up to 50,000 ⁇ -D- glucopyranose residues linked by ⁇ -1,4 and ⁇ -1,6 glycosidic bonds. Approximately 5% of the glycosidic linkages in amylopectin are ⁇ -1,6 bonds, which leads to the branched structure of the polymer.
  • Amylose and amylopectin molecules are organized into granules that are stored in plastids.
  • the starch granules produced by most plants are 15-30% amylose and 70-85% amylopectin.
  • the ratio of amylose to amylopectin and the degree of branching of amylopectin affects the physical and functional properties of the starch.
  • Functional properties, such as viscosity and stability of a gelatinized starch determine the usefulness and hence the value of starches in food and industrial applications.
  • the Rl protein of potato appears to be a granule associated enzyme that is involved in starch phosphorylation (Lorberth, R. et al. (1998) Nature Biotechnology 76:473-477). Rl activity has been associated with starch degradation in potato tubers. Studies have shown that inhibition of Rl activity leads to the synthesis of modified starch that is not degraded by enzymes present in potato tissue. If changes in starch degradation are a direct consequence of changes in the degree of phosphorylation this suggests that starch phosphorylation is an important modification that promotes degradeation.
  • nucleic acid sequences encoding all or a portion of Rl proteins in other plants would facilitate studies to better understand starch degradation and provide genetic tools for the manipulation of starch biosynthesis plant cells.
  • the instant invention relates to isolated nucleic acid fragments encoding starch Rl phosphorylation proteins. Specifically, this invention concerns isolated nucleic acid fragments encoding starch Rl phosphorylation proteins from Arabidopsis, ginger, moss, cattail, rice and soybean. In addition, this invention relates to a nucleic acid fragment that is complementary to the nucleic acid fragments encoding starch Rl phosphorylation proteins. An additional embodiment of the instant invention pertains to a polypeptide encoding all or a substantial portion of a starch Rl phosphorylation protein.
  • the instant invention relates to a chimeric gene encoding a starch Rl phosphorylation protein, or to a chimeric gene that comprises a nucleic acid fragment that is complementary to a nucleic acid fragment encoding a starch Rl phosphorylation protein, operably linked to suitable regulatory sequences, wherein expression of the chimeric gene results in production of levels of the encoded protein in a transformed host cell that is altered (i.e., increased or decreased) from the level produced in an untransformed host cell.
  • the instant invention concerns a transformed host cell comprising in its genome a chimeric gene encoding a starch Rl phosphorylation protein operably linked to suitable regulatory sequences.
  • the transformed host cell can be of eukaryotic or prokaryotic origin, and include cells derived from higher plants and microorganisms.
  • the invention also includes transformed plants that arise from transformed host cells of higher plants, and seeds derived from such transformed plants.
  • An additional embodiment of the instant invention concerns a method of altering the level of expression of a starch Rl phosphorylation protein in a transformed host cell comprising: a) transforming a host cell with a chimeric gene comprising a nucleic acid fragment encoding a starch Rl phosphorylation protein; and b) growing the transformed host cell under conditions that are suitable for expression of the chimeric gene wherein expression of the chimeric gene results in production of altered levels of starch Rl phosphorylation protein in the transformed host cell.
  • An addition embodiment of the instant invention concerns a method for obtaining a nucleic acid fragment encoding all or a substantial portion of an amino acid sequence encoding a starch Rl phosphorylation protein.
  • Figure 1 shows a comparison of the amino acid sequences of potato starch Rl phosphorylation protein set forth in NCBI Inentifier No. gi 3287270 (SEQ ID NO: 15), the instant rice Rl homolog (SEQ ID NO: 10) and the instant soybean Rl homolog (SEQ ID NO:12).
  • SEQ ID NO:l is the nucleotide sequence comprising a portion of the cDNA insert in clone acs2c.pk001.g20 encoding a portion of an Arabidopsis starch Rl phosphorylation protein.
  • SEQ ID NO:2 is the deduced amino acid sequence of a portion of a starch Rl phosphorylation protein derived from the nucleotide sequence of SEQ ID NO: 1.
  • SEQ ID NO:3 is the nucleotide sequence comprising a portion of the cDNA insert in clone ecrlc.pk007.119 encoding a portion of a ginger (Curcuma zedoari ⁇ ) starch Rl phosphorylation protein.
  • SEQ ID NO:4 is the deduced amino acid sequence of a portion of a starch Rl phosphorylation protein derived from the nucleotide sequence of SEQ ID NO:3.
  • SEQ ID NO: 5 is the nucleotide sequence comprising a portion of the cDNA insert in clone emmlc.pk001.pl 8 encoding a portion of a moss (Brachythecium oxycladon, Plagiomnium cuspidatum and Amblystegium varium) starch Rl phosphorylation protein.
  • SEQ ID NO:6 is the deduced amino acid sequence of a portion of a starch Rl phosphorylation protein derived from the nucleotide sequence of SEQ ID NO:5.
  • SEQ ID NO: 7 is the nucleotide sequence comprising a portion of the cDNA insert in clone etrlc.pk003.c21 encoding a portion of a cattail [Typha latifolia) starch Rl phosphorylation protein.
  • SEQ ID NO:8 is the deduced amino acid sequence of a portion of a starch Rl phosphorylation protein derived from the nucleotide sequence of SEQ ID NO:7.
  • SEQ ID NO:9 is the nucleotide sequence comprising a contig assembled from the cDNA inserts in clones rlm4n.pk003.pl 7, rl0n.pk088.j l 1 and rlr6.pk0099.d9 encoding a portion of a rice starch Rl phosphorylation protein.
  • SEQ ID NO: 10 is the deduced amino acid sequence of a portion of a starch Rl phosphorylation protein derived from the nucleotide sequence of SEQ ID NO:9.
  • SEQ ID NO:l 1 is the nucleotide sequence comprising a contig assembled from the cDNA inserts in clones scrlc.pk003.e3, ses4d.pk0019.b5, sll.pk0109.f9, sl2.pk0041.d7, src3c.pk006.dl 1 and src3c.pk026.j6 encoding a portion of a soybean starch Rl phosphorylation protein.
  • SEQ ID NO: 12 is the deduced amino acid sequence of a portion of a starch Rl phosphorylation protein derived from the nucleotide sequence of SEQ ID NO:l 1.
  • SEQ ID NO: 13 is the nucleotide sequence comprising a portion of the cDNA insert in clone scrlc.pk002.kl4 encoding a portion of a soybean starch Rl phosphorylation protein.
  • SEQ ID NO: 14 is the deduced amino acid sequence of a portion of a starch Rl phosphorylation protein derived from the nucleotide sequence of SEQ ID NO: 13.
  • SEQ ID NO: 15 is the amino acid sequence of potato tuber starch Rl phosphorylation protein, (NCBI Identifier No. gi 3287270).
  • the Sequence Listing contains the one letter code for nucleotide sequence characters and the three letter codes for amino acids as defined in conformity with the IUPAC-IUBMB standards described in Nucleic Acids Research 73:3021-3030 (1985) and in the Biochemical Journal 219 (No. 2 ⁇ :345-373 (1984) which are herein incorporated by reference.
  • the symbols and format used for nucleotide and amino acid sequence data comply with the rules set forth in 37 C.F.R. ⁇ 1.822.
  • an "isolated nucleic acid fragment” is a polymer of RNA or DNA that is single- or double- stranded, optionally containing synthetic, non-natural or altered nucleotide bases.
  • An isolated nucleic acid fragment in the form of a polymer of DNA may be comprised of one or more segments of cDNA, genomic DNA or synthetic DNA.
  • "contig” refers to an assemblage of overlapping nucleic acid sequences to form one contiguous nucleotide sequence. For example, several DNA sequences can be compared and aligned to identify common or overlapping regions. The individual sequences can then be assembled into a single contiguous nucleotide sequence.
  • substantially similar refers to nucleic acid fragments wherein changes in one or more nucleotide bases results in substitution of one or more amino acids, but do not affect the functional properties of the protein encoded by the DNA sequence. “Substantially similar” also refers to nucleic acid fragments wherein changes in one or more nucleotide bases does not affect the ability of the nucleic acid fragment to mediate alteration of gene expression by antisense or co-suppression technology.
  • Substantially similar also refers to modifications of the nucleic acid fragments of the instant invention such as deletion or insertion of one or more nucleotides that do not substantially affect the functional properties of the resulting transcript vis-a-vis the ability to mediate alteration of gene expression by antisense or co-suppression technology or alteration of the functional properties of the resulting protein molecule. It is therefore understood that the invention encompasses more than the specific exemplary sequences.
  • antisense suppression and co-suppression of gene expression may be accomplished using nucleic acid fragments representing less than the entire coding region of a gene, and by nucleic acid fragments that do not share 100% sequence identity with the gene to be suppressed.
  • alterations in a gene which result in the production of a chemically equivalent amino acid at a given site, but do not effect the functional properties of the encoded protein are well known in the art.
  • a codon for the amino acid alanine, a hydrophobic amino acid may be substituted by a codon encoding another less hydrophobic residue, such as glycine, or a more hydrophobic residue, such as valine, leucine, or isoleucine.
  • a codon encoding another less hydrophobic residue such as glycine
  • a more hydrophobic residue such as valine, leucine, or isoleucine.
  • changes which result in substitution of one negatively charged residue for another such as aspartic acid for glutamic acid, or one
  • substantially similar nucleic acid fragments may also be characterized by their ability to hybridize, under stringent conditions (0.1X SSC, 0.1% SDS, 65°C), with the nucleic acid fragments disclosed herein.
  • Substantially similar nucleic acid fragments of the instant invention may also be characterized by the percent similarity of the amino acid sequences that they encode to the amino acid sequences disclosed herein, as determined by algorithms commonly employed by those skilled in this art. Preferred are those nucleic acid fragments whose nucleotide sequences encode amino acid sequences that are 80% similar to the amino acid sequences reported herein. More preferred nucleic acid fragments encode amino acid sequences that are 90% similar to the amino acid sequences reported herein.
  • a "substantial portion" of an amino acid or nucleotide sequence comprises enough of the amino acid sequence of a polypeptide or the nucleotide sequence of a gene to afford putative identification of that polypeptide or gene, either by manual evaluation of the sequence by one skilled in the art, or by computer-automated sequence comparison and identification using algorithms such as BLAST (Basic Local Alignment Search Tool; Altschul, S. F., et al., (1993) J. Mol. Biol. 275:403-410; see also www.ncbi.nlm.nih.gov/BLAST/).
  • BLAST Basic Local Alignment Search Tool
  • a sequence often or more contiguous amino acids or thirty or more nucleotides is necessary in order to putatively identify a polypeptide or nucleic acid sequence as homologous to a known protein or gene.
  • gene specific oligonucleotide probes comprising 20-30 contiguous nucleotides may be used in sequence-dependent methods of gene identification (e.g., Southern hybridization) and isolation (e.g., in situ hybridization of bacterial colonies or bacteriophage plaques).
  • short oligonucleotides of 12-15 bases may be used as
  • a "substantial portion" of a nucleotide sequence comprises enough of the sequence to afford specific identification and/or isolation of a nucleic acid fragment comprising the sequence.
  • the instant specification teaches partial or complete amino acid and nucleotide sequences encoding one or more particular plant proteins. The skilled artisan, having the benefit of the sequences as reported herein, may now use all or a substantial portion of the disclosed sequences for purposes known to those skilled in this art. Accordingly, the instant invention comprises the complete sequences as reported in the accompanying Sequence Listing, as well as substantial portions of those sequences as defined above.
  • Codon degeneracy refers to divergence in the genetic code permitting variation of the nucleotide sequence without effecting the amino acid sequence of an encoded polypeptide. Accordingly, the instant invention relates to any nucleic acid fragment that encodes all or a substantial portion of the amino acid sequence encoding the starch Rl phosphorylation proteins as set forth in SEQ ID NOs:2, 4, 6, 8, 10, 12 and 14. The skilled artisan is well aware of the "codon-bias" exhibited by a specific host cell in usage of nucleotide codons to specify a given amino acid.
  • Synthetic genes can be assembled from oligonucleotide building blocks that are chemically synthesized using procedures known to those skilled in the art. These building blocks are ligated and annealed to form gene segments which are then enzymatically assembled to construct the entire gene.
  • “Chemically synthesized”, as related to a sequence of DNA, means that the component nucleotides were assembled in vitro. Manual chemical synthesis of DNA may be accomplished using well established procedures, or automated chemical synthesis can be performed using one of a number of commercially available machines.
  • the genes can be tailored for optimal gene expression based on optimization of nucleotide sequence to reflect the codon bias of the host cell.
  • the skilled artisan appreciates the likelihood of successful gene expression if codon usage is biased towards those codons favored by the host. Determination of preferred codons can be based on a survey of genes derived from the host cell where sequence information is available.
  • Gene refers to a nucleic acid fragment that expresses a specific protein, including regulatory sequences preceding (5' non-coding sequences) and following (3' non-coding sequences) the coding sequence.
  • “Native gene” refers to a gene as found in nature with its own regulatory sequences.
  • Chimeric gene refers any gene that is not a native gene, comprising regulatory and coding sequences that are not found together in nature. Accordingly, a chimeric gene may comprise regulatory sequences and coding sequences that are derived from different sources, or regulatory sequences and coding sequences derived
  • Endogenous gene refers to a native gene in its natural location in the genome of an organism.
  • a “foreign” gene refers to a gene not normally found in the host organism, but that is introduced into the host organism by gene transfer.
  • Foreign genes can comprise native genes inserted into a non-native organism, or chimeric genes.
  • a “transgene” is a gene that has been introduced into the genome by a transformation procedure.
  • Coding sequence refers to a DNA sequence that codes for a specific amino acid sequence.
  • Regulatory sequences refer to nucleotide sequences located upstream (5' non- coding sequences), within, or downstream (3' non-coding sequences) of a coding sequence, and which influence the transcription, RNA processing or stability, or translation of the associated coding sequence. Regulatory sequences may include promoters, translation leader sequences, introns, and polyadenylation recognition sequences.
  • Promoter refers to a DNA sequence capable of controlling the expression of a coding sequence or functional RNA.
  • a coding sequence is located 3' to a promoter sequence.
  • the promoter sequence consists of proximal and more distal upstream elements, the latter elements often referred to as enhancers.
  • an “enhancer” is a DNA sequence which can stimulate promoter activity and may be an innate element of the promoter or a heterologous element inserted to enhance the level or tissue-specificity of a promoter. Promoters may be derived in their entirety from a native gene, or be composed of different elements derived from different promoters found in nature, or even comprise synthetic DNA segments.
  • promoters may direct the expression of a gene in different tissues or cell types, or at different stages of development, or in response to different environmental conditions. Promoters which cause a gene to be expressed in most cell types at most times are commonly referred to as "constitutive promoters". New promoters of various types useful in plant cells are constantly being discovered; numerous examples may be found in the compilation by Okamuro and Goldberg, (1989) Biochemistry of Plants 75:1-82. It is further recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, DNA fragments of different lengths may have identical promoter activity.
  • the "translation leader sequence” refers to a DNA sequence located between the promoter sequence of a gene and the coding sequence.
  • the translation leader sequence is present in the fully processed mRNA upstream of the translation start sequence.
  • the translation leader sequence may affect processing of the primary transcript to mRNA, mRNA stability or translation efficiency. Examples of translation leader sequences have been described (Turner, R. and Foster, G. D. (1995) Molecular Biotechnology 3:225).
  • the "3' non-coding sequences" refer to DNA sequences located downstream of a coding sequence and include polyadenylation recognition sequences and other sequences encoding regulatory signals capable of affecting mRNA processing or gene expression.
  • polyadenylation signal is usually characterized by affecting the addition of polyadenylic acid tracts to the 3' end of the mRNA precursor.
  • the use of different 3' non-coding sequences is exemplified by Ingelbrecht et al., (1989) Plant Cell 7:671-680.
  • RNA transcript refers to the product resulting from RNA polymerase-catalyzed transcription of a DNA sequence. When the RNA transcript is a perfect complementary copy of the DNA sequence, it is referred to as the primary transcript or it may be a RNA sequence derived from posttranscriptional processing of the primary transcript and is referred to as the mature RNA.
  • Messenger RNA (mRNA) refers to the RNA that is without introns and that can be translated into protein by the cell.
  • cDNA refers to a double-stranded DNA that is complementary to and derived from mRNA.
  • Sense RNA transcript that includes the mRNA and so can be translated into protein by the cell.
  • Antisense RNA refers to a RNA transcript that is complementary to all or part of a target primary transcript or mRNA and that blocks the expression of a target gene (U.S. Patent No. 5,107,065, incorporated herein by reference). The complementarity of an antisense RNA may be with any part of the specific gene transcript, i.e., at the 5' non-coding sequence, 3' non-coding sequence, introns, or the coding sequence.
  • “Functional RNA” refers to sense RNA, antisense RNA, ribozyme RNA, or other RNA that may not be translated but yet has an effect on cellular processes.
  • operably linked refers to the association of nucleic acid sequences on a single nucleic acid fragment so that the function of one is affected by the other.
  • a promoter is operably linked with a coding sequence when it is capable of affecting the expression of that coding sequence (i.e., that the coding sequence is under the transcriptional control of the promoter).
  • Coding sequences can be operably linked to regulatory sequences in sense or antisense orientation.
  • expression refers to the transcription and stable accumulation of sense (mRNA) or antisense RNA derived from the nucleic acid fragment of the invention. Expression may also refer to translation of mRNA into a polypeptide.
  • Antisense inhibition refers to the production of antisense RNA transcripts capable of suppressing the expression of the target protein.
  • Overexpression refers to the production of a gene product in transgenic organisms that exceeds levels of production in normal or non-transformed organisms.
  • Co-suppression refers to the production of sense RNA transcripts capable of suppressing the expression of identical or substantially similar foreign or endogenous genes (U.S. Patent No. 5,231,020, incorporated herein by reference).
  • altered levels refers to the production of gene product(s) in transgenic organisms in amounts or proportions that differ from that of normal or non-transformed organisms.
  • “Mature” protein refers to a post-translationally processed polypeptide; i.e., one from which any pre- or propeptides present in the primary translation product have been removed.
  • Precursor protein refers to the primary product of translation of mRNA; i.e., with pre- and
  • Pre- and propeptides may be but are not limited to intracellular localization signals.
  • chloroplast transit peptide is an amino acid sequence which is translated in conjunction with a protein and directs the protein to the chloroplast or other plastid types present in the cell in which the protein is made.
  • Chloroplast transit sequence refers to a nucleotide sequence that encodes a chloroplast transit peptide.
  • a “signal peptide” is an amino acid sequence which is translated in conjunction with a protein and directs the protein to the secretory system (Chrispeels, J. J., (1991) Ann. Rev. Plant Phys. Plant Mol. Biol. 42:21-53).
  • a vacuolar targeting signal can further be added, or if to the endoplasmic reticulum, an endoplasmic reticulum retention signal (supra) may be added.
  • an endoplasmic reticulum retention signal may be added.
  • any signal peptide present should be removed and instead a nuclear localization signal included (Raikhel (1992) Plant Phys. 100:1627-1632).
  • Transformation refers to the transfer of a nucleic acid fragment into the genome of a host organism, resulting in genetically stable inheritance. Host organisms containing the transformed nucleic acid fragments are referred to as "transgenic" organisms. Examples of methods of plant transformation include Agrobacterium-mediated transformation (De Blaere et al. (1987) Meth. Enzymol. 143:211) and particle-accelerated or “gene gun” transformation technology (Klein et al. (1987) Nature (London) 327:10-13; U.S. Patent No. 4,945,050, incorporated herein by reference).
  • the nucleic acid fragments of the instant invention may be used to isolate cDNAs and genes encoding homologous proteins from the same or other plant species. Isolation of homologous genes using sequence-dependent protocols is well known in the art. Examples of sequence-dependent protocols include, but are not limited to, methods of nucleic acid hybridization, and methods of DNA and RNA amplification as exemplified by various uses of nucleic acid amplification technologies (e.g., polymerase chain reaction, ligase chain reaction).
  • genes encoding other starch Rl phosphorylation proteins could be isolated directly by using all or a portion of the instant nucleic acid fragments as DNA hybridization probes to screen libraries from any desired plant employing methodology well known to those skilled in the art.
  • Specific oligonucleotide probes based upon the instant nucleic acid sequences can be designed and synthesized by methods known in the art (Maniatis). Moreover, the entire sequences can be used directly to synthesize DNA probes by methods known to the skilled artisan such as random primer DNA labeling, nick translation, or end-labeling techniques, or RNA probes using available in vitro transcription systems.
  • primers can be designed and used to amplify a part or all of the instant sequences.
  • the resulting amplification products can be labeled directly during amplification reactions or labeled after amplification reactions, and used as probes to isolate full length cDNA or genomic fragments under conditions of appropriate stringency.
  • two short segments of the instant nucleic acid fragments may be used in polymerase chain reaction protocols to amplify longer nucleic acid fragments encoding homologous genes from DNA or RNA.
  • the polymerase chain reaction may also be performed on a library of cloned nucleic acid fragments wherein the sequence of one primer is derived from the instant nucleic acid fragments, and the sequence of the other primer takes advantage of the presence of the polyadenylic acid tracts to the 3' end of the mRNA precursor encoding plant genes.
  • the second primer sequence may be based upon sequences derived from the cloning vector.
  • the skilled artisan can follow the RACE protocol (Frohman et al., (1988) PNAS USA 55:8998) to generate cDNAs by using PCR to amplify copies of the region between a single point in the transcript and the 3' or 5' end. Primers oriented in the 3' and 5' directions can be designed from the instant sequences. Using commercially available 3' RACE or 5' RACE systems (BRL), specific 3' or 5' cDNA fragments can be isolated (Ohara et al., (1989) PNAS USA 86:5613; Loh et al., (1989) Science 243:2 ⁇ 1). Products generated by the 3' and 5' RACE procedures can be combined to generate full-length cDNAs (Frohman, M. A. and Martin, G. R., (1989) Techniques 7:165).
  • RACE protocol Frohman et al., (1988) PNAS USA 55:8998
  • Synthetic peptides representing portions of the instant amino acid sequences may be synthesized. These peptides can be used to immunize animals to produce polyclonal or monoclonal antibodies with specificity for peptides or proteins comprising the amino acid sequences. These antibodies can be then be used to screen cDNA expression libraries to isolate full-length cDNA clones of interest (Lerner, R. A. (1984) Adv. Immunol. 36:1; Maniatis).
  • nucleic acid fragments of the instant invention may be used to create transgenic plants in which the disclosed starch Rl phosphorylation proteins are present at higher or lower levels than normal or in cell types or developmental stages in which they are not normally found. This would have the effect of altering the level of starch phosphorylation in those cells.
  • Overexpression of the starch Rl phosphorylation proteins of the instant invention may be accomplished by first constructing a chimeric gene in which the coding region is operably linked to a promoter capable of directing expression of a gene in the desired tissues at the desired stage of development.
  • the chimeric gene may comprise promoter sequences and translation leader sequences derived from the same genes.
  • 3' Non- coding sequences encoding transcription termination signals may also be provided.
  • the instant chimeric gene may also comprise one or more introns in order to facilitate gene expression.
  • Plasmid vectors comprising the instant chimeric gene can then constructed.
  • the choice of plasmid vector is dependent upon the method that will be used to transform host
  • the chimeric gene described above may be further supplemented by altering the coding sequence to encode a starch Rl phosphorylation protein with appropriate intracellular targeting sequences such as transit sequences (Keegstra, K. (1989) Cell 56:247-253), signal sequences or sequences encoding endoplasmic reticulum localization (Chrispeels, J. J., (1991) Ann. Rev. Plant Phys. Plant Mol. Biol. 42:21-53), or nuclear localization signals (Raikhel, N. (1992) Plant Phys.100:1621-1632) added and/or with targeting sequences that are already present removed. While the references cited give examples of each of these, the list is not exhaustive and more targeting signals of utility may be discovered in the future.
  • a chimeric gene designed for co-suppression of the instant starch Rl phosphorylation proteins can be constructed by linking a gene or gene fragment encoding an starch Rl phosphorylation protein to plant promoter sequences.
  • a chimeric gene designed to express antisense RNA for all or part of the instant nucleic acid fragment can be constructed by linking the gene or gene fragment in reverse orientation to plant promoter sequences. Either the co-suppression or antisense chimeric genes could be introduced into plants via transformation wherein expression of the corresponding endogenous genes are reduced or eliminated.
  • the instant starch Rl phosphorylation proteins may be produced in heterologous host cells, particularly in the cells of microbial hosts, and can be used to prepare antibodies to the these proteins by methods well known to those skilled in the art.
  • the antibodies are useful for detecting starch Rl phosphorylation proteins in situ in cells or in vitro in cell extracts.
  • Preferred heterologous host cells for production of the instant starch Rl phosphorylation proteins are microbial hosts. Microbial expression systems and expression vectors containing regulatory sequences that direct high level expression of foreign proteins are well known to those skilled in the art. Any of these could be used to
  • nucleic acid fragments of the instant invention may also be used as probes for genetically and physically mapping the genes that they are a part of, and as markers for traits linked to those genes. Such information may be useful in plant breeding in order to develop lines with desired phenotypes.
  • the instant nucleic acid fragments may be used as restriction fragment length polymo ⁇ hism (RFLP) markers.
  • RFLP restriction fragment length polymo ⁇ hism
  • Southern blots Mantonis
  • the resulting banding patterns may then be subjected to genetic analyses using computer programs such as MapMaker (Lander et at., (1987) Genomics 7:174-181) in order to construct a genetic map.
  • nucleic acid fragments of the instant invention may be used to probe Southern blots containing restriction endonuclease-treated genomic DNAs of a set of individuals representing parent and progeny of a defined genetic cross. Segregation of the DNA polymorphisms is noted and used to calculate the position of the instant nucleic acid sequence in the genetic map previously obtained using this population (Botstein, D. et al., (1980) Am. J. Hum. Genet. 32:314-331).
  • Nucleic acid probes derived from the instant nucleic acid sequences may also be used for physical mapping (i.e., placement of sequences on physical maps; see Hoheisel, J. D., et al., In: Nonmammalian Genomic Analysis: A Practical Guide, Academic press 1996, pp. 319-346, and references cited therein).
  • nucleic acid probes derived from the instant nucleic acid sequences may be used in direct fluorescence in situ hybridization (FISH) mapping (Trask, B. J. (1991) Trends Genet. 7:149-154).
  • FISH direct fluorescence in situ hybridization
  • nucleic acid amplification-based methods of genetic and physical mapping may be carried out using the instant nucleic acid sequences. Examples include allele-specific amplification (Kazazian, H. H. (1989) J. Lab. Clin. Med. 114(2):95-96), polymo ⁇ hism of PCR-amplified fragments (CAPS; Sheffield, V. C. et al. (1993) Genomics 76:325-332), allele-specific ligation (Landegren, U. et al. (1988) Science 247:1077-1080), nucleotide extension reactions (Sokolov, B. P. (1990) Nucleic Acid Res. 75:3671), Radiation Hybrid Mapping (Walter, M. A.
  • Loss of function mutant pheno types may be identified for the instant cDNA clones either by targeted gene disruption protocols or by identifying specific mutants for these genes contained in a maize population carrying mutations in all possible genes (Ballinger and Benzer, (1989) Proc. Natl. Acad. Sci USA 56:9402; Koes et al., (1995) Proc. Natl. Acad. Sci USA 92:8149; Bensen et al., (1995) Plant Cell 7:75). The latter approach may be accomplished in two ways.
  • short segments of the instant nucleic acid fragments may be used in polymerase chain reaction protocols in conjunction with a mutation tag sequence primer on DNAs prepared from a population of plants in which Mutator transposons or some other mutation-causing DNA element has been introduced (see Bensen, supra).
  • the amplification of a specific DNA fragment with these primers indicates the insertion of the mutation tag element in or near the plant gene encoding the starch Rl phosphorylation protein.
  • the instant nucleic acid fragment may be used as a hybridization probe against PCR amplification products generated from the mutation population using the mutation tag sequence primer in conjunction with an arbitrary genomic site primer, such as that for a restriction enzyme site-anchored synthetic adaptor.
  • a plant containing a mutation in the endogenous gene encoding a starch Rl phosphorylation protein can be identified and obtained. This mutant plant can then be used to determine or confirm the natural function of the starch Rl phosphorylation protein gene product.
  • EXAMPLE 1 Composition of cDNA Libraries; Isolation and Sequencing of cDNA Clones cDNA libraries representing mRNAs from various Arabidopsis, ginger, moss, rice and soybean tissues were prepared. The characteristics of the libraries are described below.
  • cDNA libraries were prepared in Uni-ZAPTM XR vectors according to the manufacturer's protocol (Stratagene Cloning Systems, La Jolla, CA). Conversion of the Uni-ZAPTM XR libraries into plasmid libraries was accomplished according to the protocol provided by Stratagene. Upon conversion, cDNA inserts were contained in the plasmid vector pBluescript. cDNA inserts from randomly picked bacterial colonies containing recombinant pBluescript plasmids were amplified via polymerase chain reaction using primers specific for vector sequences flanking the inserted cDNA sequences or plasmid DNA was prepared from cultured bacterial cells.
  • Amplified insert DNAs or plasmid DNAs were sequenced in dye-primer sequencing reactions to generate partial cDNA sequences (expressed sequence tags or "ESTs"; see Adams, M. D. et al., (1991) Science 252:1651). The resulting ESTs were analyzed using a Perkin Elmer Model 377 fluorescent sequencer.
  • Example 1 The cDNA sequences obtained in Example 1 were analyzed for similarity to all publicly available DNA sequences contained in the "nr” database using the BLASTN algorithm provided by the National Center for Biotechnology Information (NCBI). The DNA sequences were translated in all reading frames and compared for similarity to all publicly available protein sequences contained in the "nr” database using the BLASTX algorithm (Gish, W. and States, D. J. (1993) Nature Genetics 3:266-272 and Altschul, Stephen F., et al. (1997) Nucleic Acids Res. 25:3389-3402) provided by the NCBI.
  • NCBI National Center for Biotechnology Information
  • the P-value (probability) of observing a match of a cDNA sequence to a sequence contained in the searched databases merely by chance as calculated by BLAST are reported herein as "pLog" values, which represent the negative of the logarithm of the reported P-value. Accordingly, the greater the pLog value, the greater the likelihood that the cDNA sequence and the BLAST "hit" represent homologous proteins.
  • SEQ ID NO: 1 the deduced amino acid sequence of this cDNA, which represents 11% of the protein (middle region), is shown in SEQ ID NO:2.
  • the sequence of a portion of the cDNA insert from clone emm lc.pk001.pl 8 is shown in SEQ ID NO:5; the deduced amino acid sequence of this cDNA, which represents 10.7% of the protein (middle region) is shown in SEQ ID NO:6.
  • the sequence of a portion of the cDNA insert from clone etrlc.pk003.c21 is shown in SEQ ID NO:7; the deduced amino acid sequence of this cDNA, which represents 7.7% of the protein (middle region), is shown in SEQ ID NO:8.
  • SEQ ID NO:9 The sequence of the rice contig composed of clones rlm4n.pk003.pl 7, rl0n.pk088.jl 1 and rlr6.pk0099.d9 is shown in SEQ ID NO:9; the deduced amino acid sequence of this contig, which represents 33% of the protein (C-terminal region) is shown in SEQ ID NO: 10.
  • Figure 1 presents an alignment of the amino acid sequence set forth in SEQ ID NO: 10 and the Solanum tuberosum sequence.
  • SEQ ID NO:l 1 The sequence of the soybean contig composed of clones scrlc.pk003.e3, ses4d.pk0019.b5, sll.pk0109.f9, sl2.pk0041.d7, src3c.pk006.dl 1 and src3c.pk026.j6 is shown in SEQ ID NO:l 1; the deduced amino acid sequence of this contig, which represents 40% of the protein (C-terminal region) is shown in SEQ ID NO: 12.
  • Figure 1 presents an alignment of the amino acid sequence set forth in SEQ ID NO: 12 and the Solanum tuberosum sequence.
  • a calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO: 12 and the Solanum tuberosum sequence revealed that the protein encoded by the contig is 76.4% similar to the Solanum tuberosum starch Rl phosphorylation protein.
  • the degree of similarity between the rice (SEQ ID NO: 10) and soybean amino acid sequences (SEQ ID NO: 12) was calculated to be 70.3% (using the Clustal algorithm).
  • sequence of a portion of the cDNA insert from clone scrlc.pk002.kl4 is shown in SEQ ID NO: 13; the deduced amino acid sequence of this cDNA, which represents 11% of the protein (middle region), is shown in SEQ ID NO: 14.
  • BLAST scores and probabilities indicate that the instant nucleic acid fragments encode portions of starch Rl phosphorylation proteins. These sequences represent the first arabidopsis, ginger, moss, cattail, rice and soybean sequences encoding starch Rl phosphorylation proteins.
  • a chimeric gene comprising a cDNA encoding a starch Rl phosphorylation protein in sense orientation with respect to the maize 27 kD zein promoter that is located 5' to the cDNA fragment, and the 10 kD zein 3' end that is located 3' to the cDNA fragment, can be constructed.
  • the cDNA fragment of this gene may be generated by polymerase chain reaction (PCR) of the cDNA clone using appropriate oligonucleotide primers. Cloning sites (Ncol or Smal) can be inco ⁇ orated into the oligonucleotides to provide proper orientation of the DNA fragment when inserted into the digested vector pML103 as described below. Amplification is then performed in a standard PCR.
  • the amplified DNA is then digested with restriction enzymes Ncol and Smal and fractionated on an agarose gel.
  • the appropriate band can be isolated from the gel and combined with a 4.9 kb Ncol-Smal fragment of the plasmid pML103.
  • Plasmid pML103 has been deposited under the terms of the Budapest Treaty at ATCC (American Type Culture Collection, 10801 University Boulevard., Manassas, VA 20110-2209), and bears accession number ATCC 97366.
  • the DNA segment from pML103 contains a 1.05 kb Sall-Ncol promoter fragment of the maize 27 kD zein gene and a 0.96 kb Smal-Sall fragment from the 3' end of the maize 10 kD zein gene in the vector pGem9Zf(+) (Promega).
  • Vector and insert DNA can be ligated at 15°C overnight, essentially as described (Maniatis). The ligated DNA may then be used to transform E. coli XL 1 -Blue (Epicurian Coli XL-1 BlueTM; Stratagene). Bacterial transformants can be screened by
  • the resulting plasmid construct would comprise a chimeric gene encoding, in the 5' to 3' direction, the maize 27 kD zein promoter, a cDNA fragment encoding a starch Rl phosphorylation protein, and the 10 kD zein 3' region.
  • the chimeric gene described above can then be introduced into corn cells by the following procedure.
  • Immature corn embryos can be dissected from developing caryopses derived from crosses of the inbred corn lines H99 and LH132.
  • the embryos are isolated 10 to 11 days after pollination when they are 1.0 to 1.5 mm long.
  • the embryos are then placed with the axis-side facing down and in contact with agarose-solidified N6 medium (Chu et al., (1975) Sci. Sin. Peking 18:659-668). The embryos are kept in the dark at 27°C.
  • Friable embryogenic callus consisting of undifferentiated masses of cells with somatic proembryoids and embryoids borne on suspensor structures proliferates from the scutellum of these immature embryos.
  • the embryogenic callus isolated from the primary explant can be cultured on N6 medium and sub-cultured on this medium every 2 to 3 weeks.
  • the plasmid, p35S/Ac (obtained from Dr. Peter Eckes, Hoechst Ag, Frankfurt, Germany) may be used in transformation experiments in order to provide for a selectable marker.
  • This plasmid contains the Pat gene (see European Patent Publication 0 242 236) which encodes phosphinothricin acetyl transferase (PAT).
  • PAT phosphinothricin acetyl transferase
  • the enzyme PAT confers resistance to herbicidal glutamine synthetase inhibitors such as phosphinothricin.
  • the pat gene in p35S/Ac is under the control of the 35S promoter from Cauliflower Mosaic Virus (Odell et al. (1985) Nature 313:810-812) and the 3' region of the nopaline synthase gene from the T-DNA of the Ti plasmid of Agrobacterium tumefaciens.
  • the particle bombardment method (Klein et al, (1987) Nature 327:70-73) may be used to transfer genes to the callus culture cells.
  • gold particles (1 ⁇ m in diameter) are coated with DNA using the following technique.
  • Ten ⁇ g of plasmid DNAs are added to 50 ⁇ L of a suspension of gold particles (60 mg per mL).
  • Calcium chloride 50 ⁇ L of a 2.5 M solution
  • spermidine free base (20 ⁇ L of a 1.0 M solution) are added to the particles.
  • the suspension is vortexed during the addition of these solutions. After 10 minutes, the tubes are briefly centrifuged (5 sec at 15,000 ⁇ m) and the supernatant removed.
  • the particles are resuspended in 200 ⁇ L of absolute ethanol, centrifuged again and the supernatant removed. The ethanol rinse is performed again and the particles resuspended in a final volume of 30 ⁇ L of ethanol.
  • An aliquot (5 ⁇ L) of the DNA-coated gold particles can be placed in the center of a KaptonTM flying disc (Bio-Rad Labs).
  • the particles are then accelerated into the corn tissue with a BiolisticTM PDS-1000/He (Bio-Rad Instruments, Hercules CA), using a helium pressure of 1000 psi, a gap distance of 0.5 cm and a flying distance of 1.0 cm.
  • the embryogenic tissue is placed on filter paper over agarose- solidified N6 medium.
  • the tissue is arranged as a thin lawn and covered a circular area of about 5 cm in diameter.
  • the petri dish containing the tissue can be placed in the chamber of the PDS-1000/He approximately 8 cm from the stopping screen.
  • the air in the chamber is then evacuated to a vacuum of 28 inches of Hg.
  • the macrocarrier is accelerated with a helium shock wave using a rupture membrane that bursts when the He pressure in the shock tube reaches 1000 psi.
  • tissue can be transferred to N6 medium that contains gluphosinate (2 mg per liter) and lacks casein or proline. The tissue continues to grow slowly on this medium. After an additional 2 weeks the tissue can be transferred to fresh N6 medium containing gluphosinate. After 6 weeks, areas of about 1 cm in diameter of actively growing callus can be identified on some of the plates containing the glufosinate- supplemented medium. These calli may continue to grow when sub-cultured on the selective medium. Plants can be regenerated from the transgenic callus by first transferring clusters of tissue to N6 medium supplemented with 0.2 mg per liter of 2,4-D. After two weeks the tissue can be transferred to regeneration medium (Fromm et al., (1990) Bio/Technology 5:833-839).
  • a seed-specific expression cassette composed of the promoter and transcription terminator from the gene encoding the ⁇ subunit of the seed storage protein phaseolin from the bean Phaseolus vulgaris (Doyle et al. (1986) J. Biol. Chem. 261 :9228-9238) can be used for expression of the instant starch Rl phosphorylation protein in transformed soybean.
  • the phaseolin cassette includes about 500 nucleotides upstream (5') from the translation initiation codon and about 1650 nucleotides downstream (3') from the translation stop codon of phaseolin. Between the 5' and 3' regions are the unique restriction endonuclease sites Nco I (which includes the ATG translation initiation codon), Sma I, Kpn I and Xba I. The entire cassette is flanked by Hind III sites.
  • the cDNA fragment of this gene may be generated by polymerase chain reaction
  • PCR PCR of the cDNA clone using appropriate oligonucleotide primers. Cloning sites can be inco ⁇ orated into the oligonucleotides to provide proper orientation of the DNA fragment when inserted into the expression vector. Amplification is then performed as described above, and the isolated fragment is inserted into a pUC18 vector carrying the seed expression cassette.
  • Soybean embroys may then be transformed with the expression vector comprising sequences encoding starch Rl phosphorylation proteins.
  • Soybean embroys may then be transformed with the expression vector comprising sequences encoding starch Rl phosphorylation proteins.
  • soybean cultivar A2872 can be cultured in the light or dark at 26°C on an appropriate agar medium for 6-10 weeks. Somatic embryos which produce secondary embryos are then excised and placed into a suitable liquid medium. After repeated selection for clusters of somatic embryos which multiplied as early, globular staged embryos, the suspensions are maintained as described below.
  • Soybean embryogenic suspension cultures can maintained in 35 mL liquid media on a rotary shaker, 150 ⁇ m, at 26°C with florescent lights on a 16:8 hour day/night schedule. Cultures are subcultured every two weeks by inoculating approximately 35 mg of tissue into 35 mL of liquid medium. Soybean embryogenic suspension cultures may then be transformed by the method of particle gun bombardment (Kline et al. (1987) Nature (London) 327:70, U.S. Patent No. 4,945,050). A DuPont BiolisticTM PDS1000/HE instrument (helium retrofit) can be used for these transformations.
  • a selectable marker gene which can be used to facilitate soybean transformation is a chimeric gene composed of the 35S promoter from Cauliflower Mosaic Virus (Odell et al. (1985) Nature 373:810-812), the hygromycin phosphotransferase gene from plasmid pJR225 (from E. coli; Gritz et al. (1983) Gene 25:179-188) and the 3' region of the nopaline synthase gene from the T-DNA of the Ti plasmid of Agrobacterium tumefaciens.
  • the seed expression cassette comprising the phaseolin 5' region, the fragment encoding the starch Rl phosphorylation protein and the phaseolin 3' region can be isolated as a restriction fragment. This fragment can then be inserted into a unique restriction site of the vector carrying the marker gene.
  • Approximately 300-400 mg of a two-week-old suspension culture is placed in an empty 60x15 mm petri dish and the residual liquid removed from the tissue with a pipette.
  • approximately 5-10 plates of tissue are normally bombarded.
  • Membrane rupture pressure is set at 1100 psi and the chamber is evacuated to a vacuum of 28 inches mercury.
  • the tissue is placed approximately 3.5 inches away from the retaining screen and bombarded three times. Following bombardment, the tissue can be divided in half and placed back into liquid and cultured as described above.
  • liquid media Five to seven days post bombardment, the liquid media may be exchanged with fresh media, and eleven to twelve days post bombardment with fresh media containing 50 mg/mL
  • EXAMPLE 6 Expression of Chimeric Genes in Microbial Cells
  • the cDNAs encoding the instant starch Rl phosphorylation proteins can be inserted into the T7 E. coli expression vector pBT430.
  • This vector is a derivative of pET-3a (Rosenberg et al. (1987) Gene 56:125-135) which employs the bacteriophage T7 RNA polymerase/T7 promoter system.
  • Plasmid pBT430 was constructed by first destroying the EcoR I and Hind III sites in pET-3a at their original positions. An oligonucleotide adaptor containing EcoR I and Hind III sites was inserted at the BamH I site of pET-3a.
  • the fragment can then be purified from the agarose gel by digestion with GELaseTM (Epicentre Technologies) according to the manufacturer's instructions, ethanol precipitated, dried and resuspended in 20 ⁇ L of water.
  • Appropriate oligonucleotide adapters may be ligated to the fragment using T4 DNA ligase (New England Biolabs, Beverly, MA).
  • T4 DNA ligase New England Biolabs, Beverly, MA
  • the fragment containing the ligated adapters can be purified from the excess adapters using low melting agarose as described above.
  • the vector pBT430 is digested, dephosphorylated with alkaline phosphatase (NEB) and deproteinized with phenol/chloroform as decribed above.
  • the prepared vector pBT430 and fragment can then be ligated at 16°C for 15 hours followed by transformation into DH5 electrocompetent cells (GIBCO BRL).
  • Transformants can be selected on agar plates containing LB media and 100 ⁇ g/mL ampicillin. Transformants containing the gene encoding the starch Rl phosphorylation protein are then screened for the correct orientation with respect to the T7 promoter by restriction enzyme analysis.
  • a plasmid clone with the cDNA insert in the correct orientation relative to the T7 promoter can be transformed into E. coli strain BL21(DE3) (Studier et al. (1986) J. Mol. Biol. 189:113-130). Cultures are grown in LB medium
  • IPTG isopropylthio- ⁇ -galactoside, the inducer
  • IPTG isopropylthio- ⁇ -galactoside, the inducer
  • incubation can be continued for 3 h at 25°.
  • Cells are then harvested by centrifugation and re-suspended in 50 ⁇ L of 50 mM Tris-HCl at pH 8.0 containing 0.1 mM DTT and 0.2 mM phenyl methylsulfonyl fluoride.
  • a small amount of 1 mm glass beads can be added and the mixture sonicated 3 times for about 5 seconds each time with a microprobe sonicator.
  • the mixture is centrifuged and the protein concentration of the supernatant determined.
  • One ⁇ g of protein from the soluble fraction of the culture can be separated by SDS-polyacrylamide gel electrophoresis. Gels can be observed for protein bands migrating at the expected molecular weight.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

Cette invention se rapporte à un fragment d'acide nucléique isolé codant une protéine R1 de phosphorylation de l'amidon, ainsi qu'à la construction d'un gène chimérique codant la totalité ou une partie de cette protéine R1 de phosphorylation de l'amidon, selon une orientation sens ou anti-sens, l'expression de ce gène chimérique entraînant la production de niveaux modifiés de cette protéine R1 de phosphorylation de l'amidon dans une cellule hôte transformée.
EP99916471A 1998-04-09 1999-04-08 Homologues de la proteine r1 de phosphorylation de l'amidon Withdrawn EP1068333A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US8114398P 1998-04-09 1998-04-09
US81143P 1998-04-09
PCT/US1999/007639 WO1999053072A1 (fr) 1998-04-09 1999-04-08 Homologues de la proteine r1 de phosphorylation de l'amidon

Publications (1)

Publication Number Publication Date
EP1068333A1 true EP1068333A1 (fr) 2001-01-17

Family

ID=22162353

Family Applications (1)

Application Number Title Priority Date Filing Date
EP99916471A Withdrawn EP1068333A1 (fr) 1998-04-09 1999-04-08 Homologues de la proteine r1 de phosphorylation de l'amidon

Country Status (4)

Country Link
EP (1) EP1068333A1 (fr)
AU (1) AU3478499A (fr)
BR (1) BR9908858A (fr)
WO (1) WO1999053072A1 (fr)

Families Citing this family (179)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100408687C (zh) 1998-11-09 2008-08-06 拜尔生物科学有限公司 来自稻米的核酸分子及其用于生产改性淀粉的用途
DE19926771A1 (de) 1999-06-11 2000-12-14 Aventis Cropscience Gmbh Nukleinsäuremoleküle aus Weizen, transgene Pflanzenzellen und Pflanzen und deren Verwendung für die Herstellung modifizierter Stärke
US6734340B2 (en) 2000-10-23 2004-05-11 Bayer Cropscience Gmbh Monocotyledon plant cells and plants which synthesise modified starch
NZ514547A (en) * 2001-09-28 2004-10-29 Duncan Stanley Plastid alph-amylase protein and nucleic acids encoding same and methods for altering the starch content of a plant
DE10208133A1 (de) * 2002-02-26 2003-09-11 Planttec Biotechnologie Gmbh Transgene Futterpflanzen mit erhöhtem Blattstärkegehalt
CL2007003743A1 (es) 2006-12-22 2008-07-11 Bayer Cropscience Ag Composicion que comprende fenamidona y un compuesto insecticida; y metodo para controlar de forma curativa o preventiva hongos fitopatogenos de cultivos e insectos.
CL2007003744A1 (es) 2006-12-22 2008-07-11 Bayer Cropscience Ag Composicion que comprende un derivado 2-piridilmetilbenzamida y un compuesto insecticida; y metodo para controlar de forma curativa o preventiva hongos fitopatogenos de cultivos e insectos.
EP1969934A1 (fr) 2007-03-12 2008-09-17 Bayer CropScience AG Phénoxyphénylamidine substituée par 4 cycloalkyl ou 4 aryl et son utilisation en tant que fongicide
EP1969929A1 (fr) 2007-03-12 2008-09-17 Bayer CropScience AG Phénylamidine substituée et son utilisation en tant que fongicide
BRPI0808786A2 (pt) 2007-03-12 2014-09-16 Bayer Cropscience Ag Di-halogenofenoxifenilamidinas e seu uso como fungicidas
BRPI0808846A2 (pt) 2007-03-12 2019-09-24 Bayer Cropscience Ag fenoxifenilamidinas 3-substituídas e seu uso como fungicidas
US8080688B2 (en) 2007-03-12 2011-12-20 Bayer Cropscience Ag 3, 4-disubstituted phenoxyphenylamidines and use thereof as fungicides
US8168567B2 (en) 2007-04-19 2012-05-01 Bayer Cropscience Ag Thiadiazolyl oxyphenyl amidines and the use thereof as a fungicide
DE102007045956A1 (de) 2007-09-26 2009-04-09 Bayer Cropscience Ag Wirkstoffkombination mit insektiziden und akariziden Eigenschaften
DE102007045920B4 (de) 2007-09-26 2018-07-05 Bayer Intellectual Property Gmbh Synergistische Wirkstoffkombinationen
DE102007045953B4 (de) 2007-09-26 2018-07-05 Bayer Intellectual Property Gmbh Wirkstoffkombinationen mit insektiziden und akariziden Eigenschaften
DE102007045919B4 (de) 2007-09-26 2018-07-05 Bayer Intellectual Property Gmbh Wirkstoffkombinationen mit insektiziden und akariziden Eigenschaften
DE102007045922A1 (de) 2007-09-26 2009-04-02 Bayer Cropscience Ag Wirkstoffkombinationen mit insektiziden und akariziden Eigenschaften
EP2090168A1 (fr) 2008-02-12 2009-08-19 Bayer CropScience AG Méthode destinée à l'amélioration de la croissance des plantes
AR069440A1 (es) * 2007-11-27 2010-01-20 Commw Scient Ind Res Org Plantas con metabolismo de almidon modificado y moleculas de acido nucleico vegetales que codifican enzimas intervinientes
EP2072506A1 (fr) 2007-12-21 2009-06-24 Bayer CropScience AG Thiazolyloxyphenylamidine ou thiadiazolyloxyphenylamidine et son utilisation en tant que fongicide
EP2168434A1 (fr) 2008-08-02 2010-03-31 Bayer CropScience AG Utilisation d'azoles destinés à l'augmentation de la résistance de plantes ou de parties de plantes contre le stress abiotique
JP2011530548A (ja) 2008-08-14 2011-12-22 バイエル・クロップサイエンス・アーゲー 殺虫性４−フェニル−１ｈ−ピラゾール類
DE102008041695A1 (de) 2008-08-29 2010-03-04 Bayer Cropscience Ag Methoden zur Verbesserung des Pflanzenwachstums
EP2201838A1 (fr) 2008-12-05 2010-06-30 Bayer CropScience AG Combinaisons utiles de matière active ayant des propriétés insecticides et acaricides
EP2198709A1 (fr) 2008-12-19 2010-06-23 Bayer CropScience AG Procédé destiné à lutter contre des parasites animaux résistants
EP2381781B1 (fr) 2008-12-29 2016-06-08 Bayer Intellectual Property GmbH Procédé pour l'utilisation améliorée d'un potentiel de production d'introduction de plantes transgéniques
EP2204094A1 (fr) 2008-12-29 2010-07-07 Bayer CropScience AG Procédé pour l'utilisation améliorée d'un potentiel de production d'introduction de plantes transgéniques
EP2223602A1 (fr) 2009-02-23 2010-09-01 Bayer CropScience AG Procédé destiné à l'utilisation améliorée du potentiel de production de plantes génétiquement modifiées
EP2039772A2 (fr) 2009-01-06 2009-03-25 Bayer CropScience AG Procédé pour améliorer l'utilisation du potentiel de production de plantes transgéniques
EP2039771A2 (fr) 2009-01-06 2009-03-25 Bayer CropScience AG Procédé pour améliorer l'utilisation du potentiel de production de plantes transgéniques
EP2039770A2 (fr) 2009-01-06 2009-03-25 Bayer CropScience AG Procédé pour améliorer l'utilisation du potentiel de production de plantes transgéniques
BRPI1006916A8 (pt) 2009-01-19 2016-05-03 Bayer Cropscience Ag Dionas cíclicas e seu uso como inseticidas, acaricidas e/ou fungicidas
EP2227951A1 (fr) 2009-01-23 2010-09-15 Bayer CropScience AG Utilisation des composés d'énaminocarbonyle de combattre des virus transmis par les insectes
PT2391608E (pt) 2009-01-28 2013-05-13 Bayer Cropscience Ag Derivados de n-cicloalquil-n-biciclíco-metilenocarboxamida fungicidas
AR075126A1 (es) 2009-01-29 2011-03-09 Bayer Cropscience Ag Metodo para el mejor uso del potencial de produccion de plantas transgenicas
EP2218717A1 (fr) 2009-02-17 2010-08-18 Bayer CropScience AG Dérivés de N-((HET)aryléthyl)thiocarboxamide fongicides
EP2398770B1 (fr) 2009-02-17 2016-12-28 Bayer Intellectual Property GmbH Dérivés de n-(phénylcycloalkyl)-carboxamide, n-(benzylcycloalkyl)-carboxamide et -thiocarboxamide fongicides
TW201031331A (en) 2009-02-19 2010-09-01 Bayer Cropscience Ag Pesticide composition comprising a tetrazolyloxime derivative and a fungicide or an insecticide active substance
EP2232995A1 (fr) 2009-03-25 2010-09-29 Bayer CropScience AG Procédé destiné à l'utilisation améliorée du potentiel de production de plantes transgéniques
CN102395271A (zh) 2009-03-25 2012-03-28 拜尔农作物科学股份公司 具有杀虫和杀螨特性的活性化合物结合物
US9012360B2 (en) 2009-03-25 2015-04-21 Bayer Intellectual Property Gmbh Synergistic combinations of active ingredients
AP3073A (en) 2009-03-25 2014-12-31 Bayer Cropscience Ag Active ingredient combinations with insecticidal and acaricidal properties
MX2011009916A (es) 2009-03-25 2011-10-06 Bayer Cropscience Ag Combinaciones de principios activos con propiedades insecticidas y acaricidas.
KR101647706B1 (ko) 2009-03-25 2016-08-11 바이엘 인텔렉쳐 프로퍼티 게엠베하 살충 및 살비성을 지니는 활성 성분 배합물
EP2239331A1 (fr) 2009-04-07 2010-10-13 Bayer CropScience AG Procédé pour améliorer l'utilisation du potentiel de production dans des plantes transgéniques
BRPI1015543A8 (pt) 2009-05-06 2016-05-24 Bayer Cropscience Ag Compostos de ciclopentanodiona e seu uso como inseticidas, acaricidas e/ou fungicidas.
EP2251331A1 (fr) 2009-05-15 2010-11-17 Bayer CropScience AG Dérivés de carboxamides de pyrazole fongicides
AR076839A1 (es) 2009-05-15 2011-07-13 Bayer Cropscience Ag Derivados fungicidas de pirazol carboxamidas
EP2255626A1 (fr) 2009-05-27 2010-12-01 Bayer CropScience AG Utilisation d'inhibiteurs de succinate déhydrogénase destinés à l'augmentation de la résistance de plantes ou de parties de plantes contre le stress abiotique
WO2010139410A2 (fr) 2009-06-02 2010-12-09 Bayer Cropscience Ag Utilisation d'inhibiteurs de la succinate déshydrogénase pour lutter contre sclerotinia ssp
MX2012000566A (es) 2009-07-16 2012-03-06 Bayer Cropscience Ag Combinaciones sinergicas de principios activos con feniltriazoles.
WO2011015524A2 (fr) 2009-08-03 2011-02-10 Bayer Cropscience Ag Dérivés d’hétérocycles fongicides
EP2292094A1 (fr) 2009-09-02 2011-03-09 Bayer CropScience AG Combinaisons de composés actifs
EP2343280A1 (fr) 2009-12-10 2011-07-13 Bayer CropScience AG Dérivés de quinoléine fongicides
MX336392B (es) 2009-12-28 2016-01-18 Bayer Cropscience Ag Derivados de hidroximoil-heterociclos fungicidas.
TW201141381A (en) 2009-12-28 2011-12-01 Bayer Cropscience Ag Fungicide hydroximoyl-tetrazole derivatives
US8796463B2 (en) 2009-12-28 2014-08-05 Bayer Cropscience Ag Fungicide hydroximoyl-tetrazole derivatives
BR112012018108A2 (pt) 2010-01-22 2015-10-20 Bayer Ip Gmbh combinações acaricidas e/ou inseticidas de ingredientes ativos
AR080443A1 (es) 2010-03-04 2012-04-11 Bayer Cropscience Ag 2-amidobencimidazoles sustituidos con fluruoalquilo
AR080827A1 (es) 2010-04-06 2012-05-09 Bayer Cropscience Ag Utilizacion del acido 4- fenil- butirico y/o de sus sales para el aumento de la tolerancia al estres en plantas
AU2011237909B2 (en) 2010-04-09 2015-08-20 Bayer Intellectual Property Gmbh Use of derivatives of the (1-cyanocyclopropyl)phenylphosphinic acid, the esters thereof and/or the salts thereof for enhancing the tolerance of plants to abiotic stress
EP2563772A1 (fr) 2010-04-28 2013-03-06 Bayer Cropscience AG Dérivés d'hydroximoyl-hétérocycles fongicides
WO2011134911A2 (fr) 2010-04-28 2011-11-03 Bayer Cropscience Ag Dérivés hydroximoyle-tétrazole fongicides
US20130116287A1 (en) 2010-04-28 2013-05-09 Christian Beier Fungicide hydroximoyl-heterocycles derivatives
US8999956B2 (en) 2010-06-03 2015-04-07 Bayer Intellectual Property Gmbh N-[(het)arylalkyl)] pyrazole(thio)carboxamides and their heterosubstituted analogues
JP5730992B2 (ja) 2010-06-03 2015-06-10 バイエル・クロップサイエンス・アーゲーＢａｙｅｒ Ｃｒｏｐｓｃｉｅｎｃｅ Ａｇ Ｎ−［（ヘタ）アリールエチル）］ピラゾール（チオ）カルボキサミド類及びそれらのヘテロ置換された類似体
UA110703C2 (uk) 2010-06-03 2016-02-10 Байєр Кропсайнс Аг Фунгіцидні похідні n-[(тризаміщений силіл)метил]-карбоксаміду
EP2580336B1 (fr) 2010-06-09 2017-05-10 Bayer CropScience NV Procédés et moyens pour modifier un génome végétal au niveau d'une séquence nucléotidique habituellement utilisée dans l'ingénierie des génomes végétaux
MX347575B (es) 2010-06-09 2017-05-03 Bayer Cropscience N V * Metodos y medios para modificar un genoma vegetal en una secuencia de nucleotidos usada comunmente en la ingenieria genetica de genomas vegetales.
JP2013532648A (ja) 2010-07-20 2013-08-19 バイエル・クロップサイエンス・アーゲー 抗菌剤としてのベンゾシクロアルケン類
EP2611300B1 (fr) 2010-09-03 2016-04-06 Bayer Intellectual Property GmbH Dérivés de dihydropyrimdinone annelés et substitués
CN103442575A (zh) 2010-09-22 2013-12-11 拜耳知识产权有限责任公司 生物或化学防治剂用于在抗性作物中防治昆虫和线虫的用途
EP2460406A1 (fr) 2010-12-01 2012-06-06 Bayer CropScience AG Utilisation de fluopyram pour contrôler les nématodes dans les cultures résistant aux nématodes
WO2012045798A1 (fr) 2010-10-07 2012-04-12 Bayer Cropscience Ag Composition fongicide comprenant un dérivé de tétrazolyloxime et un dérivé de thiazolylpipéridine
WO2012052490A1 (fr) 2010-10-21 2012-04-26 Bayer Cropscience Ag N-benzylcarboxamides hétérocycliques
CA2815114A1 (fr) 2010-10-21 2012-04-26 Juergen Benting 1-(carbonyl heterocyclique)piperidines
US9320276B2 (en) 2010-11-02 2016-04-26 Bayer Intellectual Property Gmbh N-hetarylmethyl pyrazolylcarboxamides
MX2013005258A (es) 2010-11-15 2013-07-05 Bayer Ip Gmbh N-aril pirazol(tio)carboxamidas.
MX2013005407A (es) 2010-11-15 2013-07-03 Bayer Ip Gmbh 5-halopirazolcarboxamidas.
MX2013005410A (es) 2010-11-15 2013-07-03 Bayer Ip Gmbh 5-halopirazol (tio)carboxamidas).
EP2460407A1 (fr) 2010-12-01 2012-06-06 Bayer CropScience AG Combinaisons de substance actives comprenant du pyridyléthylbenzamide et d'autres substances actives
KR20180096815A (ko) 2010-12-01 2018-08-29 바이엘 인텔렉쳐 프로퍼티 게엠베하 작물에서 선충류를 구제하고 수확량을 증가시키기 위한 플루오피람의 용도
EP2474542A1 (fr) 2010-12-29 2012-07-11 Bayer CropScience AG Dérivés fongicides d'hydroximoyl-tétrazole
EP2658853A1 (fr) 2010-12-29 2013-11-06 Bayer Intellectual Property GmbH Dérivés d'hydroxymoyl-tétrazole fongicides
EP2471363A1 (fr) 2010-12-30 2012-07-04 Bayer CropScience AG Utilisation d'acides aryl-, hétéroaryl- et benzylsulfonaminés, d'esters d'acide aminé, d'amides d'acide aminé et carbonitrile ou leurs sels pour l'augmentation de la tolérance au stress dans des plantes
EP2494867A1 (fr) 2011-03-01 2012-09-05 Bayer CropScience AG Composés substitués par un halogène en combinaison avec des fongicides
JP2014513061A (ja) 2011-03-10 2014-05-29 バイエル・インテレクチユアル・プロパテイー・ゲー・エム・ベー・ハー 処理された種子の種子安全性を保護するためのリポキト−オリゴ糖化合物の使用
US20140005230A1 (en) 2011-03-14 2014-01-02 Juergen Benting Fungicide hydroximoyl-tetrazole derivatives
US20140051575A1 (en) 2011-04-08 2014-02-20 Juergen Benting Fungicide hydroximoyl-tetrazole derivatives
AR090010A1 (es) 2011-04-15 2014-10-15 Bayer Cropscience Ag 5-(ciclohex-2-en-1-il)-penta-2,4-dienos y 5-(ciclohex-2-en-1-il)-pent-2-en-4-inos sustituidos como principios activos contra el estres abiotico de las plantas, usos y metodos de tratamiento
AR085568A1 (es) 2011-04-15 2013-10-09 Bayer Cropscience Ag 5-(biciclo[4.1.0]hept-3-en-2-il)-penta-2,4-dienos y 5-(biciclo[4.1.0]hept-3-en-2-il)-pent-2-en-4-inos sustituidos como principios activos contra el estres abiotico de las plantas
EP2511255A1 (fr) 2011-04-15 2012-10-17 Bayer CropScience AG Dérivés de prop-2-yn-1-ol et prop-2-en-1-ol substitués
AR085585A1 (es) 2011-04-15 2013-10-09 Bayer Cropscience Ag Vinil- y alquinilciclohexanoles sustituidos como principios activos contra estres abiotico de plantas
LT2997825T (lt) 2011-04-22 2019-02-25 Bayer Cropscience Aktiengesellschaft Veikliojo junginio kompozicijos, apimančios (tio)karboksamido darinį ir fungicidinį junginį
US20140173770A1 (en) 2011-06-06 2014-06-19 Bayer Cropscience Nv Methods and means to modify a plant genome at a preselected site
JP2014520776A (ja) 2011-07-04 2014-08-25 バイエル・インテレクチユアル・プロパテイー・ゲー・エム・ベー・ハー 植物における非生物的ストレスに対する活性薬剤としての置換されているイソキノリノン類、イソキノリンジオン類、イソキノリントリオン類およびジヒドロイソキノリノン類または各場合でのそれらの塩の使用
CN103717076B (zh) 2011-08-10 2016-04-13 拜耳知识产权股份有限公司 含有特定特特拉姆酸衍生物的活性化合物组合物
WO2013026740A2 (fr) 2011-08-22 2013-02-28 Bayer Cropscience Nv Procédés et moyens pour modifier un génome de plante
EP2748161A1 (fr) 2011-08-22 2014-07-02 Bayer Intellectual Property GmbH Dérivés d'hydroximoyl-tétrazole fongicides
EP2561759A1 (fr) 2011-08-26 2013-02-27 Bayer Cropscience AG 2-amidobenzimidazoles fluoroalkyl substitués et leur effet sur la croissance des plantes
CN103781353B (zh) 2011-09-09 2016-10-19 拜耳知识产权有限责任公司 用于改良植物产量的酰基高丝氨酸内酯衍生物
WO2013037717A1 (fr) 2011-09-12 2013-03-21 Bayer Intellectual Property Gmbh Dérivés de 3-{phényl[(hétérocyclylméthoxy)imino]méthyl}-1,2,4-oxadizol-5(4h)-one 4-substituée fongicides
CN103917097A (zh) 2011-09-16 2014-07-09 拜耳知识产权有限责任公司 5-苯基-或5-苄基-2-异噁唑啉-3-甲酸酯用于改善植物产量的用途
MX362112B (es) 2011-09-16 2019-01-07 Bayer Ip Gmbh Uso de fenilpirazolin-3-carboxilatos para mejorar el rendimiento de las plantas.
JP6138797B2 (ja) 2011-09-16 2017-05-31 バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングＢａｙｅｒ Ｉｎｔｅｌｌｅｃｔｕａｌ Ｐｒｏｐｅｒｔｙ ＧｍｂＨ 植物収量を向上させるためのアシルスルホンアミド類の使用
US9226505B2 (en) 2011-09-23 2016-01-05 Bayer Intellectual Property Gmbh 4-substituted 1-phenylpyrazole-3-carboxylic acid derivatives as agents against abiotic plant stress
UA115132C2 (uk) 2011-10-04 2017-09-25 Байєр Інтелекчуал Проперті Гмбх Спосіб боротьби з грибами та ооміцетами шляхом інгібування гена сахаропіндегідрогенази
WO2013050324A1 (fr) 2011-10-06 2013-04-11 Bayer Intellectual Property Gmbh Combinaison, destinée à réduire le stress abiotique de plantes, contenant de l'acide 4-phénylbutyrique (4-pba) ou un de ses sels (composant (a)) et un ou plusieurs autres composés agronomiquement actifs sélectionnés (composant(s) (b)
EP2782920B1 (fr) 2011-11-21 2016-12-21 Bayer Intellectual Property GmbH Dérivés fongicides de n-[(trisubstitué silyl)méthyl]-carboxamide
US9725414B2 (en) 2011-11-30 2017-08-08 Bayer Intellectual Property Gmbh Fungicidal N-bicycloalkyl and N-tricycloalkyl pyrazole-4-(thio)carboxamide derivatives
CN104270946B (zh) 2011-12-19 2017-05-10 拜耳农作物科学股份公司 邻氨基苯甲酸二酰胺衍生物用于防治转基因作物中的害虫的用途
JP5976837B2 (ja) 2011-12-29 2016-08-24 バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングＢａｙｅｒ Ｉｎｔｅｌｌｅｃｔｕａｌ Ｐｒｏｐｅｒｔｙ ＧｍｂＨ 殺菌性３−［（１，３−チアゾール−４−イルメトキシイミノ）（フェニル）メチル］−２−置換−１，２，４−オキサジアゾール−５（２ｈ）−オン誘導体
US9556158B2 (en) 2011-12-29 2017-01-31 Bayer Intellectual Property Gmbh Fungicidal 3-[(pyridin-2-ylmethoxyimino)(phenyl)methyl]-2-substituted-1,2,4-oxadiazol-5(2H)-one derivatives
NZ722692A (en) 2012-02-22 2018-02-23 Bayer Ip Gmbh Use of succinate dehydrogenase inhibitors (sdhis) for controlling wood diseases in grape
BR112014020898B1 (pt) 2012-02-27 2020-08-11 Bayer Intellectual Property Gmbh Combinação, método para controle de fungos fitopatogênicos prejudiciais compreendendo a referida combinação e seu uso
WO2013139949A1 (fr) 2012-03-23 2013-09-26 Bayer Intellectual Property Gmbh Compositions comprenant un composé de strigolactame pour la croissance et le rendement accrus de plantes
US9357778B2 (en) 2012-04-12 2016-06-07 Bayer Cropscience Ag N-acyl-2-(cyclo)alkypyrrolidines and piperidines useful as fungicides
US10125126B2 (en) 2012-04-20 2018-11-13 Bayer Cropscience Ag N-cycloalkyl-N-[(heterocyclylphenyl)methylene]-(thio)carboxamide derivatives
JP2015516396A (ja) 2012-04-20 2015-06-11 バイエル・クロップサイエンス・アーゲーＢａｙｅｒ Ｃｒｏｐｓｃｉｅｎｃｅ Ａｇ Ｎ−シクロアルキル−ｎ−［（三置換シリルフェニル）メチレン］−（チオ）カルボキサミド誘導体
AU2013254857B2 (en) 2012-04-23 2018-04-26 Bayer Cropscience Nv Targeted genome engineering in plants
EP2662363A1 (fr) 2012-05-09 2013-11-13 Bayer CropScience AG Biphénylcarboxamides 5-halogenopyrazoles
EP2662360A1 (fr) 2012-05-09 2013-11-13 Bayer CropScience AG Carboxamides indanyles 5-halogenopyrazoles
EP2847171A1 (fr) 2012-05-09 2015-03-18 Bayer CropScience AG 5-halogénopyrazole indanyle carboxamides
EP2662361A1 (fr) 2012-05-09 2013-11-13 Bayer CropScience AG Carboxamides indanyles de pyrazole
BR112014027643B1 (pt) 2012-05-09 2019-04-24 Bayer Cropscience Ag Pirazole-indanil-carboxamidas.
EP2662362A1 (fr) 2012-05-09 2013-11-13 Bayer CropScience AG Carboxamides indanyles de pyrazole
EP2662370A1 (fr) 2012-05-09 2013-11-13 Bayer CropScience AG Carboxamides de benzofuranyle 5-halogenopyrazole
EP2662364A1 (fr) 2012-05-09 2013-11-13 Bayer CropScience AG Carboxamides tétrahydronaphtyles de pyrazole
AR091104A1 (es) 2012-05-22 2015-01-14 Bayer Cropscience Ag Combinaciones de compuestos activos que comprenden un derivado lipo-quitooligosacarido y un compuesto nematicida, insecticida o fungicida
AU2013289301A1 (en) 2012-07-11 2015-01-22 Bayer Cropscience Ag Use of fungicidal combinations for increasing the tolerance of a plant towards abiotic stress
US20150216168A1 (en) 2012-09-05 2015-08-06 Bayer Cropscience Ag Use of substituted 2-amidobenzimidazoles, 2-amidobenzoxazoles and 2-amidobenzothiazoles or salts thereof as active substances against abiotic plant stress
MX2015004773A (es) 2012-10-19 2015-08-14 Bayer Cropscience Ag Metodo de promocion de crecimiento de planta usando derivados de carboxamida.
AU2013333916B2 (en) 2012-10-19 2017-05-25 Bayer Cropscience Ag Active compound combinations comprising carboxamide derivatives
CN105451556B (zh) 2012-10-19 2017-11-14 拜尔农科股份公司 使用羧酰胺或硫代羧酰胺衍生物处理植物以抵抗对杀真菌剂具有抗性的真菌的方法
MX2015004778A (es) 2012-10-19 2015-08-14 Bayer Cropscience Ag Metodo para mejorar la tolerancia al estres abiotico en plantas usando derivados de carboxamida o tiocarboxamida.
EP2735231A1 (fr) 2012-11-23 2014-05-28 Bayer CropScience AG Combinaisons de composés actifs
WO2014079957A1 (fr) 2012-11-23 2014-05-30 Bayer Cropscience Ag Inhibition sélective de la transduction du signal éthylène
EP2925138A1 (fr) 2012-11-30 2015-10-07 Bayer CropScience AG Mélanges fongicides et pesticides ternaires
PL2925134T3 (pl) 2012-11-30 2020-06-29 Bayer Cropscience Ag Trójskładnikowe mieszaniny grzybobójcze
CA3105365A1 (fr) 2012-11-30 2014-06-05 Bayer Cropscience Ag Melanges fongicides binaires
JP6367214B2 (ja) 2012-11-30 2018-08-01 バイエル・クロップサイエンス・アクチェンゲゼルシャフト 二成分殺菌剤混合物又は二成分殺害虫剤混合物
US9510596B2 (en) 2012-11-30 2016-12-06 Bayer Cropscience Ag Binary pesticidal and fungicidal mixtures
EP2740720A1 (fr) 2012-12-05 2014-06-11 Bayer CropScience AG Dérivés d'acides pent-2-en-4-ines bicycliques et tricycliques substitués et leur utilisation pour augmenter la tolérance au stress chez les plantes
US20150305334A1 (en) 2012-12-05 2015-10-29 Bayer Cropscience Ag Use of substituted 1-(aryl ethynyl)-, 1-(heteroaryl ethynyl)-, 1-(heterocyclyl ethynyl)- and 1-(cycloalkenyl ethynyl)-cyclohexanols as active agents against abiotic plant stress
EP2740356A1 (fr) 2012-12-05 2014-06-11 Bayer CropScience AG Dérivés d'acides (2Z)-5(1-hydroxycyclohexyl)pent-2-en-4-ines substitués
WO2014090765A1 (fr) 2012-12-12 2014-06-19 Bayer Cropscience Ag Utilisation de 1-[2-fluoro-4-méthyle-5-(2,2,2- trifluoroéthylsulfinyl)phényl]-5-amino-3-trifluorométhyl)-1 h-1,2,4 tfia zole à des fins de régulation des nématodes dans les cultures résistantes aux nématodes
AR093996A1 (es) 2012-12-18 2015-07-01 Bayer Cropscience Ag Combinaciones bactericidas y fungicidas binarias
WO2014095677A1 (fr) 2012-12-19 2014-06-26 Bayer Cropscience Ag Carboxamides difluorométhyl-nicotinique-tétrahydronaphtyle
EP2964614A1 (fr) 2013-03-07 2016-01-13 Bayer Cropscience AG Dérivés 3-{phenyl[(heterocyclylmethoxy)imino]methyl}-heterocycle fongicides
CA2908403A1 (fr) 2013-04-02 2014-10-09 Bayer Cropscience Nv Modification ciblee du genome dans des cellules eucaryotes
BR112015025637A2 (pt) 2013-04-12 2017-07-18 Bayer Cropscience Ag novos derivados de triazol
US9550752B2 (en) 2013-04-12 2017-01-24 Bayer Cropscience Aktiengesellschaft Triazolinthione derivatives
CA2909725A1 (fr) 2013-04-19 2014-10-23 Bayer Cropscience Aktiengesellschaft Procede pour l'utilisation amelioree du potentiel de production de plantes transgeniques
EP2986117A1 (fr) 2013-04-19 2016-02-24 Bayer CropScience Aktiengesellschaft Mélange insecticide ou pesticide binaire
TW201507722A (zh) 2013-04-30 2015-03-01 Bayer Cropscience Ag 做為殺線蟲劑及殺體內寄生蟲劑的ｎ－（２－鹵素－２－苯乙基）－羧醯胺類
WO2014177514A1 (fr) 2013-04-30 2014-11-06 Bayer Cropscience Ag Phénéthylcarboxamides n-substitués nématicides
BR112015031235A2 (pt) 2013-06-26 2017-07-25 Bayer Cropscience Ag derivados de n-cicloalquil-n-[(biciclil-fenil)metileno]-(tio)carboxamida
EA201600097A1 (ru) 2013-07-09 2016-06-30 Байер Кропсайенс Акциенгезельшафт Применение выбранных пиридон карбоксамидов или их солей в качестве активных веществ против абиотического стресса растений
JP6507165B2 (ja) 2013-12-05 2019-04-24 バイエル・クロップサイエンス・アクチェンゲゼルシャフト Ｎ−シクロアルキル−ｎ−｛［２−（１−置換シクロアルキル）フェニル］メチレン｝−（チオ）カルボキサミド誘導体
CN105873907B (zh) 2013-12-05 2019-03-12 拜耳作物科学股份公司 N-环烷基-n-{[2-(1-取代的环烷基)苯基]亚甲基}-(硫代)甲酰胺衍生物
AR101214A1 (es) 2014-07-22 2016-11-30 Bayer Cropscience Ag Ciano-cicloalquilpenta-2,4-dienos, ciano-cicloalquilpent-2-en-4-inas, ciano-heterociclilpenta-2,4-dienos y ciano-heterociclilpent-2-en-4-inas sustituidos como principios activos contra el estrés abiótico de plantas
AR103024A1 (es) 2014-12-18 2017-04-12 Bayer Cropscience Ag Piridoncarboxamidas seleccionadas o sus sales como sustancias activas contra estrés abiótico de las plantas
BR112017022000A2 (pt) 2015-04-13 2018-07-03 Bayer Cropscience Ag derivados de n-cicloalquil-n-(biheterocicliletileno)-(tio)carboxamida.
AU2016279062A1 (en) 2015-06-18 2019-03-28 Omar O. Abudayyeh Novel CRISPR enzymes and systems
US20190159451A1 (en) 2016-07-29 2019-05-30 Bayer Cropscience Aktiengesellschaft Active compound combinations and methods to protect the propagation material of plants
CN109715621A (zh) 2016-09-22 2019-05-03 拜耳作物科学股份公司 新的***衍生物
WO2018054829A1 (fr) 2016-09-22 2018-03-29 Bayer Cropscience Aktiengesellschaft Nouveaux dérivés de triazole et leur utilisation en tant que fongicides
US20190225974A1 (en) 2016-09-23 2019-07-25 BASF Agricultural Solutions Seed US LLC Targeted genome optimization in plants
BR112019008455A2 (pt) 2016-10-26 2019-07-09 Bayer Cropscience Ag uso de piraziflumida para o controle de sclerotinia spp. em aplicações de tratamento de semente
WO2018104392A1 (fr) 2016-12-08 2018-06-14 Bayer Cropscience Aktiengesellschaft Utilisation d'insecticides pour lutter contre les vers fil de fer
EP3332645A1 (fr) 2016-12-12 2018-06-13 Bayer Cropscience AG Utilisation de pyrimidinedione ou ses sels respectifs en tant qu'agent contre l'agression abiotique des plantes
WO2018108627A1 (fr) 2016-12-12 2018-06-21 Bayer Cropscience Aktiengesellschaft Utilisation d'indolinylméthylsulfonamides substitués ou de leurs sels pour accroître la tolérance au stress chez les plantes
WO2018204777A2 (fr) 2017-05-05 2018-11-08 The Broad Institute, Inc. Procédés d'identification et de modification d'arninc associés à des génotypes et des phénotypes cibles
WO2019025153A1 (fr) 2017-07-31 2019-02-07 Bayer Cropscience Aktiengesellschaft Utilisation de n-sulfonyl-n'-aryldiaminoalcanes et de n-sulfonyl-n'-hétéroaryldiaminoalcanes substitués ou de leurs sels pour accroître la tolérance au stress chez les plantes
US11618896B2 (en) 2017-09-21 2023-04-04 The Broad Institute, Inc. Systems, methods, and compositions for targeted nucleic acid editing
US10968257B2 (en) 2018-04-03 2021-04-06 The Broad Institute, Inc. Target recognition motifs and uses thereof
JP2021525774A (ja) 2018-06-04 2021-09-27 バイエル アクチェンゲゼルシャフトＢａｙｅｒ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ 除草活性二環式ベンゾイルピラゾール
US11384344B2 (en) 2018-12-17 2022-07-12 The Broad Institute, Inc. CRISPR-associated transposase systems and methods of use thereof

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1702518A1 (fr) * 1995-09-19 2006-09-20 Bayer BioScience GmbH Plantes synthétisant un amidon modifié, procédé de production de telles plantes, et amidon modifié obtenu à partir de ces plantes
DE19653176A1 (de) * 1996-12-19 1998-06-25 Planttec Biotechnologie Gmbh Neue Nucleinsäuremoleküle aus Mais und ihre Verwendung zur Herstellung einer modifizierten Stärke
GB9710370D0 (en) * 1997-05-20 1997-07-16 Zeneca Ltd Genetic control of fruit ripening

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9953072A1 *

Also Published As

Publication number Publication date
WO1999053072A1 (fr) 1999-10-21
BR9908858A (pt) 2000-12-19
AU3478499A (en) 1999-11-01

Similar Documents

Publication Publication Date Title
WO1999053072A1 (fr) Homologues de la proteine r1 de phosphorylation de l'amidon
US7605247B1 (en) Nucleic acid molecules encoding a wheat sucrose transporter
WO2000004166A2 (fr) Cellulose synthases vegetales
EP1097221A2 (fr) Genes codant pour des proteines d'assimilation des sulfates
EP1062353A2 (fr) Inhibiteurs des proteines d'apoptose dans des vegetaux
US7189531B2 (en) Nucleic acid encoding plant sugar transport proteins
US7332300B2 (en) Plant sugar transport proteins
US20050042722A1 (en) Plant sorbitol biosynthetic enzymes
WO1999024574A2 (fr) Homologues vegetaux de pad1 et de crm1 de levure et de jab1 humaine: regulateurs de l'activite du facteur de transcription de type ap-1
WO2000003026A2 (fr) Coactivateurs de transcription
US6893853B1 (en) Chromatin associated proteins
US7588917B2 (en) Nucleic acids encoding sugar transport proteins and methods of using same
EP1056863A1 (fr) FACTEUR DE TRANSLATION eIF-4E DE VEGETAUX
WO1999057285A2 (fr) Saccharose phosphate synthase
US20020002713A1 (en) Starch branching enzyme IIb
AU3204699A (en) Plant 1,3-beta-d-glucan synthase and brittle-1 encoding sequences
US6930226B2 (en) Granule-bound starch synthase
US6939710B2 (en) Genes encoding sulfate assimilation proteins
WO2001014540A2 (fr) Isoforme v de la synthase de l'amidon
WO1999043827A1 (fr) Cyclopropane-acyle-gras-phospholipide synthase
WO1999055870A2 (fr) Regulateurs de la transcription et de l'expression genique
WO1999053082A2 (fr) Transporteurs de l'hexose

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20000927

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): DE FR GB

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION HAS BEEN WITHDRAWN

18W Application withdrawn

Withdrawal date: 20010525