CA2404104A1 - Method for modifying lignin composition and increasing in vivo digestibility of forages - Google Patents

Method for modifying lignin composition and increasing in vivo digestibility of forages Download PDF

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Publication number
CA2404104A1
CA2404104A1 CA002404104A CA2404104A CA2404104A1 CA 2404104 A1 CA2404104 A1 CA 2404104A1 CA 002404104 A CA002404104 A CA 002404104A CA 2404104 A CA2404104 A CA 2404104A CA 2404104 A1 CA2404104 A1 CA 2404104A1
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Prior art keywords
lignification
tissue specific
specific promoter
fragment
reading frame
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CA002404104A
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French (fr)
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CA2404104C (en
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Richard A. Dixon
Dianjing Guo
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Roberts Samuels Noble Foundation Inc
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The Samuel Roberts Noble Foundation, Inc.
Richard A. Dixon
Dianjing Guo
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    • 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.)
    • 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

Abstract

Methods for transforming forage legumes or woody plants with a DNA construct comprising at least one open reading frame encoding for a caffeoyl CoA 3-O- methyltransferase enzyme or a Medicago sativa caffeic acid 3-O- methyltransferase enzyme or a fragment thereof in either a sense or antisens e orientation under a lignification-associated tissue specific promoter have been found, resulting in the down-regulation of the corresponding homologous gene either through antisense inhibition or sense suppression, as well as reduced lignin content and modified lignin composition in the transgenic plants. The expression of the caffeoyl CoA 3-O-methyltransferase transgene produces an increased syringyl lignin to guaiacyl lignin ratio in the transformed plant, and greatly improved forage digestibility.

Claims (33)

1. A method for modulating the lignin content of a forage legume comprising transforming a forage legume cell with a vector comprising a lignification-associated tissue specific promoter functionally linked to a DNA construct comprising at least one open reading frame encoding for a caffeoyl CoA 3-O-methyltransferase enzyme or a fragment thereof; and generating plants from said transformed forage legume cell
2. The method of Claim 1, wherein said forage legume cell is co-transformed with a vector comprising a lignification-associated tissue specific promoter functionally linked to a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or a fragment thereof.
3. A method for modulating the lignin content of a forage legume comprising transforming a forage legume cell with a vector comprising a lignification-associated tissue specific promoter functionally linked to a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or a fragment thereof; and generating plants from said transformed forage legume cell.
4. A method for modulating the lignin content of a forage legume comprising transforming a forage legume cell with a vector comprising a DNA construct comprising in tandem at least one open reading frame encoding for a caffeoyl CoA 3-O-methyltransferase enzyme or a fragment thereof under expression control of a first lignification-associated tissue specific promoter and at least one open reading frame encoding for a caffeic acid 3-O-methyltransferase enzyme or a fragment thereof under expression control of a second lignification-associated tissue specific promoter, wherein said first and second lignification-associated tissue specific promoter can be the same or different; and generating plants from said transformed forage legume cell.
5. A method for producing a forage legume having altered lignin composition comprising transforming a forage legume cell with a DNA construct comprising at least one open reading frame encoding for a caffeoyl CoA 3-O-methyltransferase enzyme or a fragment thereof under expression control of a lignification-associated tissue specific promoter to form a transgenic cell; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
6. The method of Claim 5, wherein said forage legume cell is co-transformed with a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or a fragment thereof under expression control of a lignification-associated tissue specific promoter.
7. A method for producing a forage legume having altered lignin composition comprising transforming a forage legume cell with a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or a fragment thereof under expression control of a lignification-associated tissue specific promoter to form a transgenic cell; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
8. A method for producing a forage legume having altered lignin composition comprising transforming a forage legume cell with a DNA construct comprising in tandem at least one open reading frame encoding for a caffeoyl CoA 3-O-methyltransferase enzyme or a fragment thereof under expression control of a first lignification-associated tissue specific promoter and at least one open reading frame encoding for a caffeic acid 3-O-methyltransferase enzyme or a fragment thereof under expression control of a second lignification-associated tissue specific promoter to form a transgenic cell, wherein said first and second lignification-associated tissue specific promoter can be the same or different;
and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
9. A method for improving the digestibility of forage legumes comprising stably incorporating into the genome of said forage legume a DNA construct comprising at least one open reading frame encoding for a 3-O-methyltransferase enzyme or a fragment thereof from the lignin biosynthetic pathway under expression control of a lignification-associated tissue specific promoter, wherein expression of said enzyme or fragment produces a change in lignin composition in said forage legume.
10. The method of Claim 9, wherein said enzyme is caffeoyl CoA 3-O-methyltransferase.
11. The method of Claim 10, wherein said forage legume is co-transformed with at least one open reading frame encoding for a caffeic acid 3-O-methyltransferase enzyme or fragment thereof under expression control of a lignification-associated tissue specific promoter.
12. The method of Claim 9, wherein said DNA construct comprising in tandem at least one open reading frame encoding for a caffeoyl CoA 3-O-methyltransferase enzyme or a fragment thereof under expression control of a first lignification-associated tissue specific promoter and at least one open reading frame encoding for a caffeic acid 3-O-methyltransferase enzyme or a fragment thereof under expression control of a second lignification-associated tissue specific promoter, wherein said first and second lignification-associated tissue specific promoter can be the same or different.
13. The method of Claim 9, 10, 11 or 12, wherein guaiacyl lignin content is reduced.
14. A method for producing a woody plant having altered lignin composition comprising transforming a woody plant cell with a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeoyl CoA 3-O-methyltransferase enzyme or fragment thereof under expression control of a lignification-associated tissue specific promoter to form a transgenic cell; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
15. The method of Claim 14, wherein said woody plant cell is co-transformed with a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or a fragment thereof under expression control of a lignification-associated tissue specific promoter.
16. A method for producing a woody plant having altered lignin composition comprising transforming a woody plant cell with a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or fragment thereof under expression control of a lignification-associated tissue specific promoter to form a transgenic cell; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
17. A method for producing a woody plant having altered lignin composition comprising transforming a woody plant cell with a DNA construct comprising in tandem at least one open reading frame encoding for a Medicago sativa caffeoyl CoA 3-O-methyltransferase enzyme or fragment thereof under expression control of a first lignification-associated tissue specific promoter and at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or fragment thereof under expression control of a second lignification-associated tissue specific promoter to form a transgenic cell, wherein said first and second lignification-associated tissue specific promoter can be the same or different; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
18. A method for modulating the lignin content of a woody plant comprising transforming a woody plant cell with a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeoyl CoA 3-O-methyltransferase enzyme or fragment thereof under expression control of a lignification-associated tissue specific promoter to form a transgenic cell; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
19. The method of Claim 18, wherein said woody plant cell is co-transformed with a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or a fragment thereof under expression control of a lignification-associated tissue specific promoter.
20. A method for modulating the lignin content of a woody plant comprising transforming a woody plant cell with a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or fragment thereof under expression control of a lignification-associated tissue specific promoter to form a transgenic cell; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
21. A method for modulating the lignin content of a woody plant comprising transforming a woody plant cell with a DNA construct comprising in tandem at least one open reading frame encoding for a Medicago sativa caffeoyl CoA 3-O-methyltransferase enzyme or fragment thereof under expression control of a first lignification-associated tissue specific promoter or fragment thereof and at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or fragment thereof under expression control of a second lignification-associated tissue specific promoter to form a transgenic cell, wherein said first and second lignification-associated tissue specific promoter can be the same or different; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
22. A method for making lignins with altered dimer bonding patterns comprising transforming a plant cell with a DNA construct comprising at least one open reading frame encoding for a caffeoyl CoA 3-O-methyltransferase enzyme or fragment thereof under expression control of a lignification-associated tissue specific promoter to form a transgenic cell; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
23. The method of Claim 22, wherein said plant cell is co-transformed with a DNA construct comprising at least one open reading frame encoding for a Medicago sativa caffeic acid 3-O-methyltransferase enzyme or a fragment thereof under expression control of a lignification-associated tissue specific promoter.
24. A method for making lignins with altered dimer bonding patterns comprising transforming a plant cell with a DNA construct comprising at least one open reading frame encoding for a caffeic acid 3-O-methyltransferase enzyme or fragment thereof under expression control of a lignification-associated tissue specific promoter to form a transgenic cell; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
25. A method for making lignins with altered dimer bonding patterns comprising transforming a plant cell with a DNA construct comprising in tandem at least one open reading frame encoding for a caffeoyl CoA 3-O-methyltransferase enzyme or fragment thereof under expression control of a first lignification-associated tissue specific promoter and at least one open reading frame encoding for a caffeic acid 3-O-methyltransferase enzyme or fragment thereof under expression control of a second lignification-associated tissue specific promoter to form a transgenic cell, wherein said first and second lignification-associated tissue specific promoter can be the same or different; and cultivating said transgenic cell under conditions conducive to regeneration and plant growth.
26. The method of Claim 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25, wherein the open reading frame is in a sense orientation.
27. The method of Claim 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25, wherein the open reading frame is in an antisense orientation.
28. The method of Claim 26, wherein said promoter is a bean PAL2 promoter.
29. The method of Claim 27, wherein said promoter is a bean PAL2 promoter.
30. A plant transformed by the method of Claim 26.
31. A plant transformed by the method of Claim 27.
32. A plant transformed by the method of Claim 28.
33. A plant transformed by the method of Claim 29.
CA2404104A 2000-03-24 2001-03-23 Method for modifying lignin composition and increasing in vivo digestibility of forages Expired - Fee Related CA2404104C (en)

Applications Claiming Priority (3)

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US19208600P 2000-03-24 2000-03-24
US60/192,086 2000-03-24
PCT/US2001/009398 WO2001073090A2 (en) 2000-03-24 2001-03-23 Method for modifying lignin composition and increasing in vivo digestibility of forages

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CA2404104A1 true CA2404104A1 (en) 2001-10-04
CA2404104C CA2404104C (en) 2012-12-11

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AR (1) AR028289A1 (en)
AU (2) AU4773101A (en)
CA (1) CA2404104C (en)
MX (1) MXPA02010432A (en)
NZ (1) NZ521545A (en)
WO (1) WO2001073090A2 (en)

Families Citing this family (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2007203378B2 (en) 2000-06-14 2011-12-08 Agriculture Victoria Services Pty Ltd Modification of lignin biosynthesis (2)
AUPR041900A0 (en) * 2000-09-29 2000-10-19 Agresearch Limited Manipulation of plant cell walls
US8921653B2 (en) 2001-06-14 2014-12-30 Agriculture Victoria Services Pty Ltd Modification of lignin biosynthesis
FR2833615A1 (en) * 2001-12-14 2003-06-20 Genoplante Valor Evaluating digestibility of fodder plants, useful for strain selection, comprises detecting alleles of the cafeoyl coenzymeA 3-O-methyltransferase gene
AUPS017402A0 (en) * 2002-01-25 2002-02-14 International Flower Developments Pty Ltd Genetic sequences and uses therefor
AU2003251522A1 (en) * 2002-06-14 2003-12-31 The Salk Institute For Biological Studies Mutant plant caffeic acid/5-hydroxyferulic acid 3/5-0-methyltransferases (comt), methods for preparation and use thereof
CA2490781A1 (en) * 2002-06-28 2004-01-08 University Of Guelph Harvest-inducible genes from alfalfa (medicago sativa) and methods of use thereof
US7329798B2 (en) 2002-06-28 2008-02-12 University Of Guelph Harvest-inducible regulatory elements and methods of using same
FR2852326B1 (en) * 2003-03-10 2007-11-30 OBTAINING PLANTS WITH IMPROVED DIGESTIBILITY HAVING INACTIVE PEROXYDASE
EP1512748A1 (en) * 2003-09-03 2005-03-09 Johnson & Johnson Research Pty Limited O-methyltransferases of tetrahydrobenzylisoquinoline alkaloid biosynthesis in papaver somniferum
CN102286481B (en) 2004-04-20 2014-07-23 辛根塔参与股份公司 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
US7663023B2 (en) 2004-07-24 2010-02-16 The Samuel Roberts Noble Foundation Modification of lignin biosynthesis
US8138392B2 (en) 2008-07-14 2012-03-20 The Samuel Roberts Noble Foundation, Inc. Disease resistant plants
US9854778B2 (en) 2011-06-30 2018-01-02 Monsanto Technology Llc Alfalfa plant and seed corresponding to transgenic event KK 179-2 and methods for detection thereof
WO2013003558A1 (en) * 2011-06-30 2013-01-03 Monsanto Technology Llc Alfalfa plant and seed corresponding to transgenic event kk 179-2 and methods for detection thereof
JP2015527878A (en) 2012-06-26 2015-09-24 モンサント テクノロジー エルエルシー Methods and compositions for improved feed quality
WO2018198049A1 (en) * 2017-04-25 2018-11-01 Cellectis Alfalfa with reduced lignin composition
CN111621520A (en) * 2020-06-22 2020-09-04 南京林业大学 Application of masson pine PmCOMT gene

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GB9119279D0 (en) * 1991-09-10 1991-10-23 Ici Plc Modification of lignin synthesis in plants
WO1994023044A1 (en) * 1993-04-02 1994-10-13 The Samuel Roberts Noble Foundation, Inc. Method for reducing ligning content in plants
CA2260907A1 (en) * 1996-07-19 1998-01-29 Purdue Research Foundation Manipulation of lignin composition in plants using a tissue-specific promoter

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NZ521545A (en) 2004-04-30
AU4773101A (en) 2001-10-08
AU2001247731B2 (en) 2006-10-12
MXPA02010432A (en) 2003-04-25
CA2404104C (en) 2012-12-11
AR028289A1 (en) 2003-04-30
WO2001073090A2 (en) 2001-10-04
WO2001073090A3 (en) 2002-05-23

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