WO1993005160A1 - Modification de la synthese de lignine dans les plantes - Google Patents

Modification de la synthese de lignine dans les plantes Download PDF

Info

Publication number
WO1993005160A1
WO1993005160A1 PCT/GB1992/001640 GB9201640W WO9305160A1 WO 1993005160 A1 WO1993005160 A1 WO 1993005160A1 GB 9201640 W GB9201640 W GB 9201640W WO 9305160 A1 WO9305160 A1 WO 9305160A1
Authority
WO
WIPO (PCT)
Prior art keywords
dna
omt
plants
lignin
plant
Prior art date
Application number
PCT/GB1992/001640
Other languages
English (en)
Inventor
Jan Van Doorsselaere
Gernard Jean Meinrad Fritig
Dirk Gustaaf Inze
Lise Jouanin
Mary Elizabeth Knight
Marc Van Montagu
Michel Legrand
Original Assignee
Zeneca Limited
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 Zeneca Limited filed Critical Zeneca Limited
Priority to CA002118793A priority Critical patent/CA2118793A1/fr
Priority to AU25167/92A priority patent/AU663726B2/en
Priority to JP5505067A priority patent/JPH06510429A/ja
Priority to EP92919119A priority patent/EP0603250A1/fr
Priority to BR9206481A priority patent/BR9206481A/pt
Priority to US08/204,288 priority patent/US5959178A/en
Publication of WO1993005160A1 publication Critical patent/WO1993005160A1/fr

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
    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against 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
    • C12N15/8255—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 lignin biosynthesis
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/1003—Transferases (2.) transferring one-carbon groups (2.1)
    • C12N9/1007—Methyltransferases (general) (2.1.1.)
    • C12N9/1011—Catechol O-methyltransferase (2.1.1.6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y201/00—Transferases transferring one-carbon groups (2.1)
    • C12Y201/01—Methyltransferases (2.1.1)
    • C12Y201/01006—Catechol O-methyltransferase (2.1.1.6)
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/11—Antisense
    • C12N2310/111—Antisense spanning the whole gene, or a large part of it

Definitions

  • This invention relates to the improvement of plants by the modification of lignin biosynthesis, particularly, but not exclusively, the improvement of digestibility of fodder crops.
  • Grassland farmers, and farmers of other fodder crops face a difficult decision each year about when to cut their crops for conservation. All grass varieties of agricultural importance suffer from the disadvantage that during the normal increase in dry matter yield with growth, the digestibility decreases. The farmer,- therefore, has, to compromise between a lower yield of highly digestible material and a higher yield of less digestible material.
  • Another limitation is that harvesting at optimum maturity may be prevented by unfavourable weather. If the decline in digestibility could be controlled or delayed, higher yields of highly digestible material could be obtained and the prevailing weather conditions would not play such a major role in determining the quality of the harvested crop.
  • Digestibility of fodder crops is determined, among other factors, by the amount and quality of lignin deposition which has taken place during growth of the plants and the degree of secondary modification of lignin deposited.
  • Beside cellulose and other poly- saccharides, lignins are an essential component of cell wall in tissues like the sclerenchyma and the xylem of vascular plants.
  • Lignins are not only important in the productivity and performance of field crops but are also of great importance in trees for paper making.
  • lignins are also used as feedstocks for the preparation of speciality chemicals such as phenolics which can be used as precursors in chemical synthesis.
  • speciality chemicals such as phenolics which can be used as precursors in chemical synthesis.
  • Lignins are the product of a dehydrogenative polymerisation of three primary precursors: the trans—coniferyl, trans-sinapyl and trans-p-coumaryl alcohols.
  • the monomers can occur in lignins in different proportions and with different types of links both with each other and with the surrounding cell wall polysaccharides,thus producing a wide variety of polymers.
  • These polymers, or "lignin cores" are always associated covalently with hemicelluloses. Most lignins also contain varying amounts of aromatic carboxylic acids in ester-like combinations.
  • lignin monomers are a part of the phenylpropanoid biosynthesis pathway, which is also responsible for the production of a wide range of compounds including flavonoid pigments, isoflavonoids, coumarin phytoalexins and cell division promoting dehydrodiconiferyl glucosides. Phenylalanine is deaminated to produce cinnamic acid.
  • OMT O-methyl transferase
  • the resulting two phenolics, ferulic acid and sinapic acid, respectively, are the precursors of coniferyl alcohol and sinapyl alcohol which are together with coumaryl alcohol substrates for peroxidases (Lewis and Yamamoto, 1990).
  • Lignification takes place during this phase. It starts in the cell corners and extends along the middle lamella, through the primary wall and, finally, to the secondary wall. External factors can induce qualitative and quantitative modifications in lignification.
  • the synthesis of new types of lignins, sometimes in tissues which are not normally lignified, may be induced by infection with pathogenic microorganisms. Lignification is stimulated by light, as well as by low calcium levels, by boron, by mechanical stress and by infection.
  • OMTs O-methyltransferase ⁇
  • OMT I used mainly caffeic acid and 5-hydroxyferulic acid as a substrate and is the OMT actively present in healthy plants.
  • OMT II and OMT III have a broader substrate specificity and also use catechol as substrate.
  • TMV Upon infection with TMV, an increase in activity of all three OMTs was shown. Based on this observation, it has been postulated that the OMT I is specifically involved in lignification, whereas OMT II and OMT III have a.
  • lignin may have a negative effect on plant growth.
  • crops such as wheat, oilseed rape, sugar beet or maize might presumably increase the grain yield.
  • Trees with reduced lignin contents or altered lignin structure will lead to a reduction in the cost of the paper as less lignin will have to be removed during the pulping process.
  • novel papers may be produced due to the purity of cellulose fibre which could otherwise not be produced.
  • Reduction of lignification can be achieved by the application of chemical inhibitors to plants.
  • a more effective method controlling lignin deposition and structure is the inhibition of expression of the CAD gene using antisense RNA.
  • Antisense RNA technology is an appropriate molecular biology approach to the inhibition of lignification.
  • An antisense RNA is an RNA produced by the transcription of the non-coding DNA strand (nonsense).
  • antisense RNA has the same sequence as the coding DNA strand and is complementary to the mRNA product of a specific gene.
  • RNA sequence which is complementary to a sequence of bases in a mRNA: complementary in the sense that each base (or a majority of bases) in the antisense sequence (read in the 3' to 5' sense) is capable of pairing with the corresponding base (G with C, A with U) in the mRNA sequence read in the 5' to 3' sense.
  • RNA Ribonucleic acid
  • antisense RNA may be produced in the cell by transformation with an appropriate DNA construct arranged to transcribe backwards part of the coding strand (as opposed to the template strand) of the relevant gene (or of a
  • An object of the present invention is to provide plants having an altered ability to synthesi ⁇ e lignin.
  • the DNA insert contained in the clones pPLC4 and pTOMTI and variants thereof such as are permitted by the degeneracy of the genetic code or the functional equivalents thereof.
  • the present invention provides a reco binant DNA construct containing the said DNA under control of a transcriptional control sequence operative in plants, so that the construct can generate mRNA in plant cells.
  • the aforesaid DNA is in antisense orientation.
  • the aforesaid DNA is in sense orientation thus to provide one or more additional copies of the said DNA in the plant genome.
  • the present invention provides DNA constructs comprising a transcriptional initiation region operative in plants positioned for transcription of a DNA sequence encoding RNA complementary to a substantial run of bases showing substantial homology to an mRNA encoding the protein produced by the gene in pPLC4 pTOMTl.
  • the invention further provides plant cells, and plants derived therefrom having stably incorporated in their genomes the aforesaid DNA in sense or antisense orientation, and fruit and seeds of such plants.
  • the present invention is principally concerned with the suppression of lignin formation and, that being so, the inserted gene will be in antisense orientation, but there are instances where over-production of lignin may have an advantageous effect, for example to improve plant stalk strength and resistance to diseases, and the present invention provides means for achieving amplification of the lignin biosynthetic ability of plants.
  • the invention relates generally to the regulation of the plant's lignin biosynthetic pathway, in which OMT plays a dominant role, in order that the production of OMT, and hence the production and composition of lignin is altered by insertion of the OMT gene, or a portion thereof
  • constructs of the invention may be inserted into plants to regulate the production of the CAD enzyme. Depending on the nature of the construct, the production of the protein may be increased, or reduced, either throughout or at particular stages in the life of the plant. It is also possible to target the expression of the gene to a specific cell types of the plant, such as the epidermis, the xylem, the roots etc.
  • DNA constructs according to the invention preferably comprise a sequence of at least 50 bases which is homologous to the DNA of the insert in pPLC4 or pOMTl.A and pOMTl.B There is no theoretical upper limit to the base sequence - it may be as long as the relevant mRNA produced by the cell - but for convenience it will generally be found suitable to use sequences between 100 and 1000 bases in length. The preparation of such constructs is described in more detail below.
  • the preferred source of antisense RNA for use in the present invention is DNA derived from the clone pPLC4 or pOMTl.A and pOMTl.B
  • the required DNA encoding antisense RNA can be obtained in several ways: by cutting an appropriate sequence of DNA from pPLC4 or pOMTl.A or pOMTl.B (or any other source of the OMT gene); by synthesising a DNA fragment using synthetic oligonucleotides which are annealed and then ligated together in such a way as to give suitable restriction sites at each end; by using synthetic oligonucleotides in a polymerase chain reaction (PCR) to generate the required fragment with suitable restriction sites at each end.
  • PCR polymerase chain reaction
  • the DNA is then cloned into a vector containing upstream promoter and downstream terminator sequences, the cloning being carried out so that the DNA sequence is inverted with respect to its orientation to the promoter in the strand from which it was cut.
  • the strand that was formerly the template strand becomes the coding strand, and vice versa.
  • the new vector will thus encode RNA in a base sequence which is complementary to the sequence of pPLC4 and pTOMTl mRNAs.
  • the two RNA strands are complementary not only in their base sequence but also in their orientations (5' to 3 r ).
  • pPLC4 As source of the DNA base sequence for transcription, it is convenient to use a cDNA clone such as pPLC4.
  • the base sequence of pPLC4 is shown in Figure 1 and the base sequences of pOMTl.A and pOMTl.B is shown in Figure 2.
  • the clone pPLC4 has been deposited at the National Collections of Industrial and Marine Bacteria, PO Box 31, of 23 St Machar Drive, Aberdeen AB2 1RY, Scotland, as a plasmid in E.coli, strain 'sure', under the reference NCIB 40436 on August 22, 1991.
  • the clone pTOMTl.A has been deposited at the National Collections of Industrial and Marine Bacteria, PO Box 31, of 23 St Machar Drive,
  • a source of DNA for the base sequence for transcription is the promoter of the OMT gene itself or other genes involved in lignification such as the promoter of the phenyl alanine ammonia lyase gene or its modified version which permits expression in xylem tissue, or the s-Adenosyl methionine synthase gene or the promoter of the extensin gene.
  • Such a gene may differ from the cDNA of pPLC4 and pOMTlA or pOMTl.B in that introns may be present. The introns are not transcribed into mRNA (or, if so transcribed, are subsequently cut out).
  • a further way of obtaining a suitable DNA base sequence for transcription is to synthesise it ab initio from the appropriate bases, for example using Figure 1 and Figure 2 as a guide.
  • Recombinant DNA and vectors according to the present invention may be made as follows.
  • a suitable vector containing the desired base sequence for transcription for example pPLC4 and pOMTl.A and pOMTl.B
  • restriction enzymes to out cut the sequence.
  • the DNA strand so obtained is cloned (in reverse orientation) into a second vector containing the desired promoter sequence
  • Agrobacterium tumefaciens nopaline synthase gene Agrobacterium tumefaciens nopaline synthase gene.
  • constitutive promoters such as cauliflower mosaic virus 35S RNA
  • inducible or developmentally regulated promoters such as the PAL gene promoter
  • Use of a constitutive promoter will tend to affect functions in all parts of the plant: while by using a tissue specific promoter, functions may be controlled more selectively.
  • tissue-specific promoter has the advantage that the antisense or sense RNA is only produced in the tissue in which its action is required.
  • Vectors according to the invention may be used to transform plants as desired, to make plants according to the invention.
  • Dicotyledonous plants such as alfalfa, oil seed rape etc, may be transformed by Agrobacterium Ti plasmid technology, for example as described by Bevan (1984) Nucleic Acid Research, 12, 8711-8721. Such transformed plants may be replicated sexually, or by cell or tissue culture.
  • Poplar and aspen transformation using Agrobacterium tumefaciens can be performed as described by De Block [Plant Physiol. (1990) 93:1110-1116].
  • Stem internode pieces are used as a tissue source for incubation with an Agrobacterium tumefaciens strain (C58CRif (pMP90.) harbouring chimeric marker genes (bar/neo) on its non- oncogenic T-DNA.
  • C58CRif pMP90.
  • chimeric marker genes bar/neo
  • transgenic shoots were obtained 3 months and 6 months after incubation, respectively.
  • RNA in the plant cells can be controlled by suitable choice of promoter sequences, or by selecting the number of copies, or the site of integration, of the DNA sequences according to the invention that are introduced into the plant genome. In this way it may be possible to modify lignification to a greater or lesser extent.
  • the constructs of our invention may be used to transform cells of both monocotyledonous and dicotyledonous plants in various ways known to the art. In many cases such plant cells (particularly when they are cells of dicotyledonous plants) may be cultured to regenerate whole plants which subsequently reproduce to give successive generations of genetically modified plants. Examples of genetically modified plants according to the present invention include, alfalfa, oil seed rape, sunflower, sorghum, maize, festuca,and trees such as eucalyptus, poplar, and pine.
  • antisense RNA in order to determine the phenotype of transgenic plants which show modified, that is increased or reduced, expression of pPLC4 or pTOMTl by the use of antisense and sense expression vectors.
  • Figure 1 shows the complete nucleotide sequence and deduced amino acid sequence from pPLC4.
  • Figure 2 shows the combined nucleotide sequence and deduced amino acid sequence of pOMTl.A and pOMTl.B.
  • Figure 3 shows the nucleotide sequence of a complete cDNA clone of OMTI from a stem tobacco library.
  • Figure 4 is the nucleotide sequence of a OMT
  • Figure 5 shows the construction of antisense and sense vectors using the 5' end 500 bp and the 3' end 900 bp BamHl fragments from pPLC4.
  • Figure 6 shows the construction of an antisense vector using a 1.4 kb PCR fragment containing the complete pPLC4 clone.
  • Table 1 shows the OMT activity using caffeic acid as a substrate in young leaves and xylem tissue from poplar trees (Populus trichocarpa x P. deltaides) .
  • OMT activity was purified both from leaves and xylem tissue.
  • the procedure for the purification of the OMTs was established by Dumas et al., (1988).
  • the purification of OMT activity from a total protein extract of poplar leaves via ammonium sulphate precipitation, desalting on Sephadex G25, Q- Sepharose chromatography, and adenosine agarose affinity chromatography, and finally, MonoQ chromatography resulted in one 38-kDa protein. About 5 ⁇ g of OMT was obtained from 100 g of leaves.
  • Antibodies raised against OMT I and OMT II from tobacco were used to test for cross-reactions with the poplar OMTs. Antibodies raised against
  • the specific activity of the purified OMT towards three different O-diphenolic substrates was measured. Using catechol, caffeic acid, and hydroxyferulic acid as substrate, we found an OMT activity of 0, 30, and 15 nkat/mg protein, respectively.
  • OMT 1 from tobacco uses mainly caffeic acid and hydroxyferulic acid as a substrate (Collendavelloo et al., 1981).
  • the purified poplar OMT has an OMT
  • the 38—kDa protein isolated from poplar xylem tissue was digested with trypsin and the peptides were separated on reverse-phase HPLC.
  • Four peptides were sequenced: peptide 45 (R/KDLPHVIEDAPSYGVEHVGGDMF) peptide 49 (LVDVGGGTGAW) peptide 51 (GINFDLPHVIEDAP) peptide 52 (VILVE?ILPVAPD) .
  • peptide 45 is a mixture of two peptides with arginine and lysine, respectively, as first amino acid. Trypsin cleaves proteins after a lysine or arginine residue, except when this is followed by proline, glutamic acid, or aspartic acid. This implies that the first amino acid preceding the sequence of peptide 45 has to be either an arginine or a lysine.
  • the sequences of peptide 45 and 51 are overlapping.
  • the cDNA insert is 1375 nucleotides in length and contains one open reading-frame of 1092 nucleotides encoding a protein of 364 amino acids (calculated M 39,720; Pi 5.45).
  • the pPCL2 clone contains a cDNA of 1,420 nucleotides and contains also one ORF of 1,092 nucleotides encoding a protein of 364 amino acids. There is only a 3-amino acid difference between the proteins of the pPCL4 and the pPCL2 clones. Amino acids 97, 191, and 361 in the protein of pPCL2 are leucine, isoleucine, and phenylalanine, respectively. Thus a mixture of at least three closely related isoforms, as shown by the two isoforms present in peptide 45 and by the amino acid sequence of peptide 51 which is entirely found back in the deduced OMT sequence. EXAMPLE 4
  • RNA gel blots were performed.
  • the antisense B RNA was detected (strongly) in candidate ASB 5B and candidates ASB 3Am ASB 5A and ASB 7A.
  • the antisense A RNA levels were high in candidates ASA LA and candidates ASA 5B, ASA 17A,
  • ASA 6A and ASA 2B No antisense RNA was detected in transgenic plants containing the pGSJ780A T-DNA.
  • the difference in the antisense RNA levels can be explained by position effects.
  • the sense B RNA can be detected easily in candidates SB 5A and SB 8A and not in candidates SB 2B, SB 4A, SB 7A and SB 10A.
  • the sense A RNA amount is high in candidates
  • OMT activity was measured in different organelle ⁇ /tissues (Table ⁇ 3 and 4). Two candidates (p35SASOM3B 4A and 6A) with a lower OMT activity in petioles, xylem and phloem (two to three times lower) in comparison to wild type and control plants, were identified.
  • Lignin analysis of the tran ⁇ genic poplars The transgenic plants were analysed for their lignin composition (Table 5). Lignin is a complex polymer of three different units: p-hydroxyphenyl (H), guaiacyl (G) and syringyl (S) monomers. Poplar lignin contain ⁇ both guaiacyl and syringyl units. A typical syringyl/guaiacyl (S/G) ratio for poplar i ⁇ 2/1. Due to the anti ⁇ ense inhibition we expect that the monomer ratio will be modified, resulting in a lower S/G ratio. Lignin characterisation of xylem was performed by the analysis of degradation products recovered from thioacidolysis. This method allows the estimation of the different units, involved in the lignin characteristic structure.
  • a tobacco OMTI cDNA clones were isolated from a leaf cDNA library prepared from RNA of TMV infected tobacco leaves.
  • the ⁇ gtll phages were plated on Escherichia coli Y1090 and fusion proteins induced with lO M
  • the longe ⁇ t clone was 614 bp long and was submitted to phage amplification and DNA purification by CsCl gradient (4).
  • the 614 bp cDNA insert was subloned by EcoRI restriction digestion in pBluescript KS(+) (Stratagene, Inc). According to standard methods (4), deletion from both the extremities of the plasmid were generated by ExoIII-Mung Bean digestions and the primers and the T7 DNA polymerase.
  • EXAMPLE 7 Construction and screening of a ⁇ ZapII cDNA library from tobacco leaf RNA and characterisation of a complete clone
  • Poly(A)+RNA from 48 hours TMV infected leaves was used to construct an oligo(dT) primed cDNA.
  • Double-stranded cDNA was ligated to hemi-phosphorylated EcoR I/Not I adaptors (Pharmacia), ligated into ⁇ ZapII vector (Stratagene, Inc) and packaged using Gigapack in vitro packaging extracts (Stratagene, Inc).
  • the resulting cDNA library titled 1.8 X 10 7 pfu//g cDNA.
  • the o3.614 clone was used as DNA probe.
  • the 614 bp DNA fragment was purified from purified from 0.8% agarose gel (PrepAGene, BIORAD), 32p labelled by random oligonucleotide-primed synthesis and used to screen a ⁇ ZapII library made by standard protocols (4). After three cycles of screening. twelve positive clones were isolated from approximately 1.8 X 10 plaque-forming units inserts from positive phage were rescued as Bluescript plasmids by R408 helper phage mediated in vivo excision, as described by the manufacture
  • OMT I mRNAs were localised in parenchyma cells of xylem and phloem, with a marked signal around the nuclei.
  • OMT I mRNAs were found to accumulate particularly in the upper and lower epidermis in a ring of tissue surrounding TMV-induced necrotic lesions, were no cell-type specific hybridisation was found in the healthy leaf.
  • EXAMPLE 9 Design of antisense vectors
  • the full-length OMT clone was cloned in the vector pGSJ780A in antisense direction by PCR yielding plasmid p35SASOM3C ( Figure 6).
  • the correct direction of the inserts has been confirmed by sequencing.
  • the vector pGSJ780A is a binary vector with the pVSl origin of replication and a Sm/Sp resistance gene for selection in Agrobacterium tumefaciens. Between the T-DNA borders there is a nos-nptll-ocs cassette and a multiple cloning site) .
  • the construction of the tobacco antisense vector follows that described for the poplar vectors.
  • the insert was inserted into the vector PGSJ780A.

Landscapes

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

Abstract

On obtient la régulation de la biosynthèse de la lignine dans les plantes par insertion dans le génome de la plante en modifiant la capacité de la plante à synthésiser l'enzyme O-méthyle-transférase, une enzyme intervenant dans le processus de biosynthèse de la lignine. On peut améliorer la production d'O-méthyle-transférase par insertion dans le génome de la plante, en transformant une ou plusieurs copies supplémentaires du gène de l'O-méthyle-transférase ou on peut inhiber la production de cette enzyme en introduisant un gène codant pour un ARNm antisens dirigé contre l'ARNm codé par le gène d'O-méthyle-transférase endogène.
PCT/GB1992/001640 1991-09-10 1992-09-09 Modification de la synthese de lignine dans les plantes WO1993005160A1 (fr)

Priority Applications (6)

Application Number Priority Date Filing Date Title
CA002118793A CA2118793A1 (fr) 1991-09-10 1992-09-09 Modification de la synthese de la lignine chez les plantes
AU25167/92A AU663726B2 (en) 1991-09-10 1992-09-09 Modification of lignin synthesis in plants
JP5505067A JPH06510429A (ja) 1991-09-10 1992-09-09 植物におけるリグニン合成の変性
EP92919119A EP0603250A1 (fr) 1991-09-10 1992-09-09 Modification de la synthese de lignine dans les plantes
BR9206481A BR9206481A (pt) 1991-09-10 1992-09-09 DNA codificando o-metil transferase de ácido caféico DNA recombinante célula de planta planta derivada da mesma e processo para a regulação de biosíntese de lignina numa planta
US08/204,288 US5959178A (en) 1991-10-09 1992-09-09 Modification of lignin synthesis in plants

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB919119279A GB9119279D0 (en) 1991-09-10 1991-09-10 Modification of lignin synthesis in plants
GB9119279.9 1991-09-10

Publications (1)

Publication Number Publication Date
WO1993005160A1 true WO1993005160A1 (fr) 1993-03-18

Family

ID=10701156

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1992/001640 WO1993005160A1 (fr) 1991-09-10 1992-09-09 Modification de la synthese de lignine dans les plantes

Country Status (7)

Country Link
EP (1) EP0603250A1 (fr)
JP (1) JPH06510429A (fr)
AU (1) AU663726B2 (fr)
BR (1) BR9206481A (fr)
CA (1) CA2118793A1 (fr)
GB (1) GB9119279D0 (fr)
WO (1) WO1993005160A1 (fr)

Cited By (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994023044A1 (fr) * 1993-04-02 1994-10-13 The Samuel Roberts Noble Foundation, Inc. Procede de reduction de la teneur en lignine dans des plantes
US5451514A (en) * 1991-04-26 1995-09-19 Zeneca Limited Modification of lignin synthesis in plants
WO1997045549A1 (fr) * 1996-05-31 1997-12-04 Centre National De La Recherche Scientifique Sequences d'adn codant pour des laccases, et leurs applications dans le domaine de la regulation des teneurs en lignines des plantes
WO1998011205A2 (fr) * 1996-09-11 1998-03-19 Genesis Research & Development Corporation Limited Materiaux et procedes permettant de modifier le contenu de plantes en lignine
WO1998050570A2 (fr) * 1997-05-08 1998-11-12 The Regents Of The University Of Michigan Procedes et compositions ayant trait a l'utilisation d'(iso)eugenol methyltransferase
US5866791A (en) * 1995-10-25 1999-02-02 Zeneca Limited Modification of lignin synthesis in plants
US5886243A (en) * 1995-11-30 1999-03-23 Board Of Control Of Michigan Technological University Genetic engineering of wood color in plants
US5922928A (en) * 1995-11-30 1999-07-13 Board Of Control Of Michigan Technological University Genetic transformation and regeneration of plants
WO2000037662A2 (fr) * 1998-12-21 2000-06-29 E.I. Du Pont De Nemours And Company Promoteur de s-adenosyl-l-methionine synthetase et son utilisation pour exprimer des genes transgeniques chez des vegetaux
WO2000056897A1 (fr) * 1999-03-22 2000-09-28 Rhobio Promoteur inductible comtii, gene chimere le comprenant et plantes transformees
FR2791359A1 (fr) * 1999-03-22 2000-09-29 Rhone Poulenc Agrochimie Promoteur inductible comtii, gene chimere le comprenant et plantes transformees
WO2001007472A1 (fr) * 1999-07-23 2001-02-01 Dcv, Inc. Facteur d'activation de la cytokine hautement purifie et procedes d'utilisation
US6204434B1 (en) 1996-09-11 2001-03-20 Genesis Research & Development Corporation Limited Materials and methods for the modification of plant lignin content
WO2001073090A2 (fr) * 2000-03-24 2001-10-04 The Samuel Roberts Noble Foundation, Inc. Procede de modification d'une composition de lignine et augmentation de la digestibilite in vivo des fourrages
WO2002009501A1 (fr) * 2000-07-28 2002-02-07 Molecular Plant Brdding Nominees Ltd. Modification de la resistance de plantes a des maladies et/ou a des parasites
WO2002026994A1 (fr) * 2000-09-29 2002-04-04 Agriculture Victoria Services Pty Ltd Manipulation de parois de cellules vegetales
US6410718B1 (en) 1996-09-11 2002-06-25 Genesis Research & Development Corporation Ltd. Materials and methods for the modification of plant lignin content
US6441272B1 (en) 1998-12-02 2002-08-27 The University Of Georgia Research Foundation, Inc. Modification of lignin content and composition in plants
US6455762B1 (en) 1997-11-12 2002-09-24 Board Of Control Of Michigan Technological University Methods of modifying lignin in plants by transformation with a 4-coumarate coenzyme a ligase nucleic acid
WO2003018819A1 (fr) * 2001-08-31 2003-03-06 Genoplante-Valor Procede d'obtention de plants de maïs transformes a caracteristiques de digestibilite ameliorees, plants de maïs obtenus par le procede et utilisations
WO2003078629A1 (fr) 2002-03-20 2003-09-25 Basf Plant Science Gmbh Produit de synthese et procede de regulation de l'expression genique
US6703539B1 (en) * 1998-01-22 2004-03-09 National Research Council Of Canada Method and compositions for modifying levels of secondary metabolic compounds in plants
US7087426B2 (en) 1996-09-11 2006-08-08 Agrigenesis Biosciences Ltd. Materials and methods for the modification of plant lignin content
US7217858B2 (en) 1998-12-21 2007-05-15 E. I. Du Pont De Nemours And Company S-adenosyl-L-methionine synthetase promoter and its use in expression of transgenic genes in plants
EP2267138A2 (fr) 1998-04-08 2010-12-29 Commonwealth Scientific and Industrial Research Organization Procédés et moyens d'obtention de phénotypes modifiés
US7888553B2 (en) 2000-03-24 2011-02-15 The Samuel Roberts Noble Foundation Method for modifying lignin composition and increasing in vivo digestibility of forages
US7910326B2 (en) 1996-09-11 2011-03-22 Arborgen, Inc. Materials and methods for the modification of plant lignin content
US8013214B2 (en) 2004-09-29 2011-09-06 Biogemma S.A.S. Maize having an improved digestibility
US8115054B2 (en) 2006-10-24 2012-02-14 Biogemma Maize with enhanced tolerance to fungal pathogen
US8129588B2 (en) 2004-04-20 2012-03-06 Syngenta Participations Ag Regulatory sequences for expressing gene products in plant reproductive tissue
US8198511B2 (en) 2007-07-31 2012-06-12 Biogemma Maize having improved digestibility
US8536406B2 (en) 2008-04-28 2013-09-17 Michigan Technological University COMT1 gene fiber-specific promoter elements from poplar
WO2013184768A1 (fr) 2012-06-05 2013-12-12 University Of Georgia Research Foundation, Inc. Compositions et méthodes d'inactivation génique dans les plantes
US9238818B2 (en) 2004-04-20 2016-01-19 Syngenta Participations Ag Methods and genetic constructs for modification of lignin composition of corn cobs
EP2980220A1 (fr) 2005-09-20 2016-02-03 BASF Plant Science GmbH Procédés améliorés de contrôle de l'expression de gènes

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2005597A1 (fr) * 1988-12-15 1990-06-15 Wolfgang W. Schuch Plantes renfermant une quantite moindre de lignine ou de la lignine alteree

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2005597A1 (fr) * 1988-12-15 1990-06-15 Wolfgang W. Schuch Plantes renfermant une quantite moindre de lignine ou de la lignine alteree

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
BIOLOGICAL ABSTRACTS vol. 73 , 1982, Philadelphia, PA, US; abstract no. 64323, MARIGO, G., ET AL. 'Biosynthesis of carbon-14-labeled ferulic acid and sinapic acid' *
BIOLOGICAL ABSTRACTS vol. 84 , 1987, Philadelphia, PA, US; abstract no. 19249, HERMANN, C., ET AL. 'Enzymatic synthesis of lignin: Purification to homogeneity of the three O-methyltransferases of tobacco and production of specific antibodies' *
BIOLOGICAL ABSTRACTS vol. 94 , 1992, Philadelphia, PA, US; abstract no. 89217, JAECK, E., ET AL. 'Regulation of enzymes involved in lignin biosynthesis: Induction of O-methyltransferase mRNAs during the hypersensitive reaction of tobacco to tobacco mosaic virus' *
J. CELL. BIOCHEM. SUPPL., MEETING HELD APRIL 10-16, 1992. vol. 16F, 1992, page 219 NI, W., ET AL. 'Modification of lignin biosynthesis by genetic manipulation of caffeic acid O-methyltransferase' *
J. CELL. BIOCHEM. SUPPL.,MEETING HELD JAN 10-17, 1991. vol. 15A, 1991, page 144 CAMPBELL, W.H., ET AL. 'Seasonal expression of a lignin specific O-methyltransferase cloned from aspen developing secondary xylem' *
PLANT PHYSIOLOGY, (SUPPL.) vol. 93, no. 1, May 1990, ROCKVILLE, MD, USA. page 15 BUGOS, R.C., ET AL. 'Cloning and expression of lignin bispecific O-methyltransferase' *
PLANT PHYSIOLOGY, (SUPPL.) vol. 99, no. 1, May 1992, ROCKVILLE, MD, USA. page 19 PODILA, G.K., ET AL. 'Antisense expression of an aspen O-methyltransferase construct in transgenic tobacco via Agrobacterium' *
PLANT PHYSIOLOGY. vol. 96, no. 2, June 1991, ROCKVILLE, MD, USA. pages 577 - 583 LAGRIMINI, L.M., ET AL. 'Wound-induced deposition of polyphenols in transgenic plants overexpressing peroxidase' *
PLANT PHYSIOLOGY. vol. 97, no. 1, September 1991, ROCKVILLE, MD, USA. pages 7 - 14 GOWRI, G., ET AL. 'Stress responses in Alfalfa (Medicago sativa L.)' *
PLANT PHYSIOLOGY. vol. 98, no. 2, February 1992, ROCKVILLE, MD, USA. pages 796 - 797 DUMAS, B., ET AL. 'Nucleotide sequence of a complementary DNA encoding O-methyltransferase from poplar' *

Cited By (60)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5451514A (en) * 1991-04-26 1995-09-19 Zeneca Limited Modification of lignin synthesis in plants
US6066780A (en) * 1991-04-26 2000-05-23 Zeneca Limited Modification of lignin synthesis in plants
WO1994023044A1 (fr) * 1993-04-02 1994-10-13 The Samuel Roberts Noble Foundation, Inc. Procede de reduction de la teneur en lignine dans des plantes
US5866791A (en) * 1995-10-25 1999-02-02 Zeneca Limited Modification of lignin synthesis in plants
US5922928A (en) * 1995-11-30 1999-07-13 Board Of Control Of Michigan Technological University Genetic transformation and regeneration of plants
US5886243A (en) * 1995-11-30 1999-03-23 Board Of Control Of Michigan Technological University Genetic engineering of wood color in plants
WO1997045549A1 (fr) * 1996-05-31 1997-12-04 Centre National De La Recherche Scientifique Sequences d'adn codant pour des laccases, et leurs applications dans le domaine de la regulation des teneurs en lignines des plantes
FR2749322A1 (fr) * 1996-05-31 1997-12-05 Centre Nat Rech Scient Sequences d'adn codant pour des laccases, et leurs applications dans le domaine de la regulation des teneurs en lignines des plantes
WO1998011205A2 (fr) * 1996-09-11 1998-03-19 Genesis Research & Development Corporation Limited Materiaux et procedes permettant de modifier le contenu de plantes en lignine
US5850020A (en) * 1996-09-11 1998-12-15 Genesis Research & Development Corporation, Ltd. Materials and method for the modification of plant lignin content
US5952486A (en) * 1996-09-11 1999-09-14 Genesis Research & Development Corporation Limited Materials and methods for the modification of plant lignin content
WO1998011205A3 (fr) * 1996-09-11 1998-08-20 Genesis Res & Dev Corp Ltd Materiaux et procedes permettant de modifier le contenu de plantes en lignine
US7087426B2 (en) 1996-09-11 2006-08-08 Agrigenesis Biosciences Ltd. Materials and methods for the modification of plant lignin content
US6410718B1 (en) 1996-09-11 2002-06-25 Genesis Research & Development Corporation Ltd. Materials and methods for the modification of plant lignin content
US7910326B2 (en) 1996-09-11 2011-03-22 Arborgen, Inc. Materials and methods for the modification of plant lignin content
US6204434B1 (en) 1996-09-11 2001-03-20 Genesis Research & Development Corporation Limited Materials and methods for the modification of plant lignin content
WO1998050570A3 (fr) * 1997-05-08 1999-02-04 Univ Michigan Procedes et compositions ayant trait a l'utilisation d'(iso)eugenol methyltransferase
WO1998050570A2 (fr) * 1997-05-08 1998-11-12 The Regents Of The University Of Michigan Procedes et compositions ayant trait a l'utilisation d'(iso)eugenol methyltransferase
US6455762B1 (en) 1997-11-12 2002-09-24 Board Of Control Of Michigan Technological University Methods of modifying lignin in plants by transformation with a 4-coumarate coenzyme a ligase nucleic acid
US6969784B2 (en) 1997-11-12 2005-11-29 Board Of Control Of Michigan Technological University Genetic engineering of plants through manipulation of lignin biosynthesis
US6831208B1 (en) 1997-11-12 2004-12-14 Board Of Control Of Michigan Technological University 4-coumarate co-enzyme a ligase promoter
US6703539B1 (en) * 1998-01-22 2004-03-09 National Research Council Of Canada Method and compositions for modifying levels of secondary metabolic compounds in plants
EP3214177A2 (fr) 1998-04-08 2017-09-06 Commonwealth Scientific and Industrial Research Organisation Procédés et moyens pour obtenir des phénotypes modifiés
EP2267139A2 (fr) 1998-04-08 2010-12-29 Commonwealth Scientific and Industrial Research Organization Procédés ét moyens d'obtention de phénotypes modifies
EP2267138A2 (fr) 1998-04-08 2010-12-29 Commonwealth Scientific and Industrial Research Organization Procédés et moyens d'obtention de phénotypes modifiés
US6441272B1 (en) 1998-12-02 2002-08-27 The University Of Georgia Research Foundation, Inc. Modification of lignin content and composition in plants
EP1967588A3 (fr) * 1998-12-21 2008-10-29 E.I. Du Pont De Nemours And Company Promoteur de la S-adénosy-L-méthionine synthétase et son utilisation dans l'expression de gènes transgéniques chez les plantes
US7217858B2 (en) 1998-12-21 2007-05-15 E. I. Du Pont De Nemours And Company S-adenosyl-L-methionine synthetase promoter and its use in expression of transgenic genes in plants
US7834242B2 (en) 1998-12-21 2010-11-16 E. I. Du Pont De Nemours And Company S-adenosyl-L-methionine synthetase promoter and its use in expression of transgenic genes in plants
US7741537B2 (en) 1998-12-21 2010-06-22 E.I. Du Pont De Nemours And Company S-adenosyl-L-methionine synthetase promoter and its use in expression of transgenic genes in plants
US8426680B2 (en) 1998-12-21 2013-04-23 E I Du Pont De Nemours And Company S-adenosyl-L-methionine synthetase promoter and its use in expression of transgenic genes in plants
WO2000037662A3 (fr) * 1998-12-21 2000-10-26 Du Pont Promoteur de s-adenosyl-l-methionine synthetase et son utilisation pour exprimer des genes transgeniques chez des vegetaux
WO2000037662A2 (fr) * 1998-12-21 2000-06-29 E.I. Du Pont De Nemours And Company Promoteur de s-adenosyl-l-methionine synthetase et son utilisation pour exprimer des genes transgeniques chez des vegetaux
WO2000056897A1 (fr) * 1999-03-22 2000-09-28 Rhobio Promoteur inductible comtii, gene chimere le comprenant et plantes transformees
FR2791360A1 (fr) * 1999-03-22 2000-09-29 Aventis Cropscience Sa Promoteur inductible, comtii, gene chimere le comprenant et plantes transformees
FR2791359A1 (fr) * 1999-03-22 2000-09-29 Rhone Poulenc Agrochimie Promoteur inductible comtii, gene chimere le comprenant et plantes transformees
JP4763193B2 (ja) * 1999-07-23 2011-08-31 アルキオン ライフ サイエンシズ リミテッド ライアビリティ カンパニー 高度に精製されたサイトカイン活性化因子および使用方法
JP2003506019A (ja) * 1999-07-23 2003-02-18 ディーシーヴィー インコーポレイテッド 高度に精製されたサイトカイン活性化因子および使用方法
WO2001007472A1 (fr) * 1999-07-23 2001-02-01 Dcv, Inc. Facteur d'activation de la cytokine hautement purifie et procedes d'utilisation
WO2001073090A2 (fr) * 2000-03-24 2001-10-04 The Samuel Roberts Noble Foundation, Inc. Procede de modification d'une composition de lignine et augmentation de la digestibilite in vivo des fourrages
WO2001073090A3 (fr) * 2000-03-24 2002-05-23 Samuel Roberts Noble Found Inc Procede de modification d'une composition de lignine et augmentation de la digestibilite in vivo des fourrages
AU2001247731B2 (en) * 2000-03-24 2006-10-12 The Samuel Roberts Noble Foundation, Inc. Method for modifying lignin composition and increasing in vivo digestibility of forages
US7888553B2 (en) 2000-03-24 2011-02-15 The Samuel Roberts Noble Foundation Method for modifying lignin composition and increasing in vivo digestibility of forages
US7122718B2 (en) 2000-07-28 2006-10-17 Dairy Australia Limited Modification of plant resistance to diseases and/or pests
WO2002009501A1 (fr) * 2000-07-28 2002-02-07 Molecular Plant Brdding Nominees Ltd. Modification de la resistance de plantes a des maladies et/ou a des parasites
WO2002026994A1 (fr) * 2000-09-29 2002-04-04 Agriculture Victoria Services Pty Ltd Manipulation de parois de cellules vegetales
WO2003018819A1 (fr) * 2001-08-31 2003-03-06 Genoplante-Valor Procede d'obtention de plants de maïs transformes a caracteristiques de digestibilite ameliorees, plants de maïs obtenus par le procede et utilisations
FR2829151A1 (fr) * 2001-08-31 2003-03-07 Agronomique Inst Nat Rech Procede d'obtention de plants de mais transformes a caracteristiques de digestibilite ameliorees, plants de mais obtenus par le procede et utilisations
WO2003078629A1 (fr) 2002-03-20 2003-09-25 Basf Plant Science Gmbh Produit de synthese et procede de regulation de l'expression genique
US8129588B2 (en) 2004-04-20 2012-03-06 Syngenta Participations Ag Regulatory sequences for expressing gene products in plant reproductive tissue
US9238818B2 (en) 2004-04-20 2016-01-19 Syngenta Participations Ag Methods and genetic constructs for modification of lignin composition of corn cobs
US8597913B2 (en) 2004-04-20 2013-12-03 Syngenta Participations Ag Method of constructing an expression cassette comprising regulatory sequences of a target gene of a plant for expressing gene products
US8679844B2 (en) 2004-04-20 2014-03-25 Syngenta Participations Ag MADS gene regulatory sequences for expressing gene products in plant reproductive tissue
US8013214B2 (en) 2004-09-29 2011-09-06 Biogemma S.A.S. Maize having an improved digestibility
EP2980220A1 (fr) 2005-09-20 2016-02-03 BASF Plant Science GmbH Procédés améliorés de contrôle de l'expression de gènes
US8115054B2 (en) 2006-10-24 2012-02-14 Biogemma Maize with enhanced tolerance to fungal pathogen
US8330006B2 (en) 2006-10-24 2012-12-11 Biogemma Maize with good digestibility and disease resistant
US8198511B2 (en) 2007-07-31 2012-06-12 Biogemma Maize having improved digestibility
US8536406B2 (en) 2008-04-28 2013-09-17 Michigan Technological University COMT1 gene fiber-specific promoter elements from poplar
WO2013184768A1 (fr) 2012-06-05 2013-12-12 University Of Georgia Research Foundation, Inc. Compositions et méthodes d'inactivation génique dans les plantes

Also Published As

Publication number Publication date
BR9206481A (pt) 1995-10-31
AU2516792A (en) 1993-04-05
GB9119279D0 (en) 1991-10-23
JPH06510429A (ja) 1994-11-24
AU663726B2 (en) 1995-10-19
CA2118793A1 (fr) 1993-03-18
EP0603250A1 (fr) 1994-06-29

Similar Documents

Publication Publication Date Title
AU663726B2 (en) Modification of lignin synthesis in plants
EP0584117B1 (fr) Modification de la synthese de la lignine dans les plantes
US5648210A (en) Nucleotide sequences of galactinol synthase from zucchini and soybean
JP4871487B2 (ja) 植物におけるリグニン含量及び組成、及びセルロース含量の同時制御方法
US8173866B1 (en) Modulation of plant xylan synthases
AU705401B2 (en) DNA sequences coding for a cynnamoyl CoA reductase and their uses in the field of regulating the lignin levels of plants
CN101410515A (zh) 调节植物木质素的组合物和方法
AU2007308770B2 (en) Method for modifying plant architecture and enhancing plant biomass and/or sucrose yield
NZ531985A (en) Manipulation of flavonoid biosynthesis in Trifolium repens (white clover), Lolium perenne (ryegrass) and Festuca (fescue)
US5959178A (en) Modification of lignin synthesis in plants
US10428343B2 (en) Modification of lignin biosynthesis via sense suppression
CA2005597A1 (fr) Plantes renfermant une quantite moindre de lignine ou de la lignine alteree
US5866791A (en) Modification of lignin synthesis in plants
Hayakawa et al. Molecular cloning and tissue-specific expression of two genes that encode caffeic acid O-methyltransferases from Populus kitakamiensis
CA2415554A1 (fr) Procede permettant d'influencer la teneur en sinapine de plantes et cellules vegetales transgeniques
Marillia et al. Molecular cloning of a Brassica napus thiohydroximate S‐glucosyltransferase gene and its expression in Escherichia coli
WO1999001558A1 (fr) Genes et polypeptides de plantes et leurs utilisations
Chiang et al. Methods for simultaneous control of lignin content and composition, and cellulose content in plants
Mittasch Hydroxycinnamate glycosyltransferase genes in Brassica napus, encoding key enzymes in sinapate ester metabolism
WO2009104181A1 (fr) Plantes ayant une teneur en lignine génétiquement modifiée et leurs procédés de production
US20030073828A1 (en) Epimerase gene and use thereof
Allina Phenylpropanoid metabolism in poplar: characterization of the 4-coumarate: CoA ligase gene family and a putative hydroxycinnamoyl-CoA thioesterase
AU2010258280A1 (en) Production of plants with reduced lignin content
AU2008201994A1 (en) Manipulation of flavonoid biosynthesis in plants (2)

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU BB BG BR CA CS FI HU JP KP KR LK MG MN MW NO PL RO RU SD US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL SE BF BJ CF CG CI CM GA GN ML MR SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
COP Corrected version of pamphlet

Free format text: PAGES 3/12,6/12 AND 11/12,DRAWINGS,REPLACED BY NEW PAGES BEARING THE SAME NUMBER

WWE Wipo information: entry into national phase

Ref document number: 1992919119

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2118793

Country of ref document: CA

WWP Wipo information: published in national office

Ref document number: 1992919119

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 08204288

Country of ref document: US

WWW Wipo information: withdrawn in national office

Ref document number: 1992919119

Country of ref document: EP