WO1994023044A1 - Procede de reduction de la teneur en lignine dans des plantes - Google Patents

Procede de reduction de la teneur en lignine dans des plantes Download PDF

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Publication number
WO1994023044A1
WO1994023044A1 PCT/US1994/003356 US9403356W WO9423044A1 WO 1994023044 A1 WO1994023044 A1 WO 1994023044A1 US 9403356 W US9403356 W US 9403356W WO 9423044 A1 WO9423044 A1 WO 9423044A1
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Prior art keywords
plants
alfalfa
lignin
leu
gene
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PCT/US1994/003356
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English (en)
Inventor
Richard A. Dixon
Weiting Ni
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The Samuel Roberts Noble Foundation, Inc.
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Publication date
Application filed by The Samuel Roberts Noble Foundation, Inc. filed Critical The Samuel Roberts Noble Foundation, Inc.
Priority to AU66211/94A priority Critical patent/AU6621194A/en
Publication of WO1994023044A1 publication Critical patent/WO1994023044A1/fr

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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
    • 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.)

Definitions

  • This invention relates to a method for reducing the lignin content in plants using an antisense gene for caffeic acid 3-O-methyltransferase from alfalfa.
  • Lignin is an insoluble polymer which occurs in the secondary thickening of plant cell walls and is primarily responsible for the rigidity of plant stems. Although lignin is essential for vascular function in plants, and may be involved in disease resistance in cereals, there is much interest in producing plants with reduced lignin content. Lignin residues are a problem in the paper processing industry. Further, the digestibility of forage grasses by cattle decreases with increasing lignin content. Lignin concentration has been reported to be the single most important measurable factor limiting the in vitro digestibility of other constituents including cellulose, hemicellulose, and neutral detergent fiber. easier, "In vitro Digestibility of Dry Matter in Cell Wall Constituents of Smooth Bromegra ⁇ s Forage," Crop Sci . , Vol. 27, pp. 931-934, 1987. easier further reported that small increases (approximately 1%) in lignin content can result in relatively large decreases (approximately 7%) in digestibility of plant dry matter.
  • U.S. Patent No. 5,107,065 issued to Shewmaker, et al., on April 21, 1992, describes the regulation of gene expression in plant cells using antisense regulation.
  • Antisense regulation involves the integration of a gene under the transcriptional control of a promoter which is functional in the host and in which the strand to be transcribed is complementary to the strand of DNA that is normally transcribed from the endogenous genes one wishes to regulate. The integrated gene is referred to as the antisense gene.
  • U.S. Patent No. 5,107,065 describes methods and compositions for modulating R A utilization, particularly modulation of a phenotypic property of a plant host cell.
  • the patent discusses that the complementary sequence can be at least about 15 nucleotides in length, usually being fewer than about 5,000 nucleotides. According to the patent, the particular sites to which the sequence binds, and the length of the sequence, need to be determined based empirically on the experience observed with a particular sequence. Specifically, the patent describes the use of antisense technology in the regulated modulation of the expression of polygalacturonase in tomatoes. The patent describes that the ability to reduce the production of polygalacturonase could have a positive effect on the solids content of the tomato plant and improve tomato processing.
  • recombinant DNA comprising a nucleotide sequence encoding mRNA which is substantially homologous or complementary to RNA encoded by an endogenous plant gene, or part thereof, which encodes an enzyme essential to lignin biosynthesis is provided so that mRNA transcribed from the insert inhibits the production of the enzyme from the endogenous gene.
  • Enzymes essential to lignin biosynthesis include cinnamyl alcohol dehydrogenase (CAD) , cinnamoyl-CoA reductase (CCR) , and catechol-O-methyltransferase (synonymous with caffeic acid 3-O-methyltransferase, COMT) .
  • CAD cinnamyl alcohol dehydrogenase
  • CCR cinnamoyl-CoA reductase
  • catechol-O-methyltransferase catechol-O-methyltransferase
  • Podila, et al. reported on the use of antisense technology in constructing transgenic tobacco plants utilizing antisense expression of an aspen xylem-specific O-methyltransferase (OMT).
  • OMT O-methyltransferase
  • the pre ⁇ ent invention relates to a method for reducing the lignin content of plants comprising in ⁇ erting a tran ⁇ cribable anti ⁇ en ⁇ e gene for caffeic acid 3-O-methyltran ⁇ fera ⁇ e of alfalfa into the genome of a plant.
  • the pre ⁇ ent invention in another a ⁇ pect, relates to a reagent for transfecting plants to reduce the lignin content thereof comprising a transcribable antisense gene for caffeic acid 3-O-methyltransferase of alfalfa.
  • FIGURE 1 depict ⁇ the vector con ⁇ truction ⁇ trategy for tobacco transfection.
  • FIGURE 2 depicts the lignin content of transfected tobacco plants compared with controls.
  • FIGURE 3 is a southern blot of tran ⁇ genic tobacco geno ic DNA.
  • FIGURE 4 depict ⁇ the COMT activity of transfected tobacco plants and a control.
  • FIGURE 5 depicts the vector construction strategy for alfalfa transfection.
  • FIGURE 6 depict ⁇ the lignin content and COMT activity of tran ⁇ fected alfalfa plants compared with controls.
  • FIGURE 7a depicts the complementary sequence of the fragment (highlighted) of the gene utilized in the anti ⁇ ense vector construct for tobacco.
  • FIGURE 7b depicts the complementary ⁇ equence of the fragment (highlighted) of the gene utilized in the anti ⁇ ense vector construct for alfalfa.
  • FIGURE 8 depicts the lignin biosynthesi ⁇ pathway.
  • Pillonel, et al. reported that plants exhibiting a reduction in lignin content exhibited a decrease in products of CAD. As can be seen in FIGURE 7, CAD (9) is involved near the end of the biosynthetic pathway for lignin. Therefore, modification of lignin content through down-regulation of CAD using antisense technology has been attempted. Contrastingly, Pillonel, et al. reported an increase in products of COMT involved earlier in the biosynthetic pathway of lignin (4) . Nonetheless, once COMT cDNA was cloned, studies were undertaken to use antisense COMT constructs to modify the lignin content of plants.
  • the method according to the invention is carried out in forage plants.
  • the same protocol could be applied to other forage legumes and grasse ⁇ , for which transformation/regeneration sy ⁇ tem ⁇ are available.
  • FIGURE 1 shows the strategies used for vector construction.
  • a 1.3 kb fragment of alfalfa COMT cDNA, pCOMT in FIGURE 1 was digested with various restriction enzymes, and released fragments were inserted into the vector pRTL2 in the orientation indicated by the arrows.
  • the vector pRTL2 has a CaMV 35S promoter which can drive the expression of the inserted DNA fragment, but the vector is incapable of incorporating the DNA fragment into the targeted plant genome.
  • the cas ⁇ ette containing the 35S promoter and the inserted DNA fragment was thus removed from pRTL2 by J ⁇ indlll digestion and inserted into the binary vector pGA482 for insertion into the targeted plant genome.
  • All elements between BL (border left) and BR (border right) in pGA482 can be inserted into the target plant genome by AgroJba ⁇ terium-mediated transformation. . Any suitable binary plant transformation vector can be used.
  • the neomycin phosphotransfera ⁇ e (NPT II) segment in pGA482 was used as a selectable marker, since it confers kanamycin resistance on plants during transformation and regeneration.
  • FIGURES 1 and 5 "B” repre ⁇ ent ⁇ a BamHl re ⁇ triction ⁇ ite and "B , " repre ⁇ ent ⁇ a BelI re ⁇ triction ⁇ ite.
  • the relevant re ⁇ triction enzyme sites on the pCOMT sequence are depicted in FIGURE 7a.
  • the portion of the pCOMT sequence used for the tobacco construct is highlighted in FIGURE 7a and was essentially the sequence located between the two Bell restriction enzyme sites depicted in FIGURE 7a. Insertion of this portion of the sequence in inverted orientation into vector pRTL2 ensured transcription of the complementary, antisen ⁇ e ⁇ trand.
  • the orientation of the anti ⁇ ense gene was determined by restriction enzyme analysi ⁇ using standard procedures as described by Sambrook, et al., Molecular Cloning: A Laboratory Manual , Cold Spring Harbor Lab, 2d ed., 1989, incorporated herein by reference.
  • Leaf disk transformation procedures (Rogers, et al., Methods in Enzymology, Vol. 118, pp. 627-640, 1986, incorporated herein by reference) were used for tobacco (Nicotiana tabacum cv. Xanthi) transformation.
  • the binary vector pGA482 was mobilized into Agrobacterium tumefaciens strain LBA4404 using the freeze-thaw method described by An, et al., Plant Molecular Biolo ⁇ v Manual. Kluwer Academic Publishing, Netherlands, 1988 (incorporated herein by reference) .
  • a single colony of transformed bacteria was inoculated in Murashige and Skoog (MS) medium (Mura ⁇ hige and Skoog, Physiologia Plantarum, Vol.
  • Transgenic plants exhibited no morphological difference ⁇ from control tobacco plant ⁇ under normal growth chamber and greenhouse conditions. Control tobacco plants were regenerated from leaf disk ⁇ without transformation with foreign genes.
  • Alteration in the lignin biosynthetic pathway may also change the content and composition of soluble phenolic compounds (e.g., decrease metabolites derived from the COMT reaction) .
  • Phenolic analysis was performed as described in Elkind, et al., Proc. Natl . Acad. Sci . USA, Vol. 87, pp. 9057-9061, 1990 (incorporated herein by reference) .
  • Tobacco stems were cut in half and upper portions were homogenized with a mortar and pestle, extracted with water once, and then extracted with methanol for at least two days with ⁇ everal change ⁇ of methanol. After extraction, debris (cell wall material) was saved for lignin analysis (see below) .
  • Dried cell wall material (0.1 gm) was incubated in 5 ml 2N HC1 containing 10% (v/v) thioglycolic acid at 95°C for 2 hours. After centrifugation, the acidic supernatant was removed and the pellet was suspended in 5 ml of 0.5 N NaOH for at least 20 hours with gentle shaking.
  • the residue was dissolved in methanol and analyzed by HPLC. The results indicated that the lignin composition was ⁇ imilar in the control ⁇ and the transgenic plants. There was no major alteration in relative levels of the components (i.e., sinapyl, coniferyl, and coumary1 units, see FIGURE 8) .
  • Plant tissues were homogenized in 100 mM Tris-HCl, pH 7.5, 2 mM EDTA and 5 mM
  • the assay reaction was initiated by adding the sub ⁇ trate ⁇ caffeic acid and [methyl- 14 C]S- adenosyl-L-methionine to 150 ⁇ l of plant extract, and the reaction mixture was incubated at 37°C for 30 minutes.
  • the as ⁇ ay ⁇ were ⁇ topped with 30 ⁇ l of 1 N HCl, and labeled products were partitioned into 250 ⁇ l of ethyl acetate: hexane (1:1, v/v).
  • 150 ⁇ l of the organic phase was analyzed by scintillation counting. The results are depicted in FIGURE 4.
  • Six of the seven transgenic plants tested exhibited from 20% to 60% of the average control value.
  • the control value is the average of three independent untran ⁇ formed plant ⁇ plus plant B107, which was transformed but does not contain the antisense gene ( ⁇ ee FIGURE 3) .
  • the pre ⁇ ence of the COMT anti ⁇ ense construct in transgenic plants was determined by Southern blot analysis.
  • Total DNA isolated from tobacco leaves (Junghans, et al., Biotechnique ⁇ , Vol. 8, p. 176, 1990, incorporated herein by reference) was dige ⁇ ted with the re ⁇ triction enzyme Hindlll, resolved on an agarose gel and blotted onto a nylon membrane. Blot hybridization and other standard manipulations were as described by Sambrook, et al., Molecular Cloning: A Laboratory Manual , Cold Spring Harbor Lab, 2d ed. , 1989, incorporated herein by reference. The results are depicted in FIGURE 3.
  • transgenic plants except B107 contained an approximately 1.4 kb fragment hybridizing to the pCOMT sequence.
  • a COMT antisen ⁇ e construct was successive ⁇ fully introduced into transgenic tobacco plants. All of the seven transgenic plants analyzed showed NPT II activity indicating that these plants were truly transformed with the construct. Six of the seven plants exhibited various ranges of COMT enzyme activities, which were 20 to 60% of the COMT activities in control plants. Histochemical staining for lignin in the stems of the transgenic plants revealed a significant reduction of lignin in six of the seven transgenic plants. Southern blot analysis (FIGURE 3) indicated that the transgenic plant (B107) with normal lignin content (compared to control plants) did not contain the COMT antisen ⁇ e fragment.
  • the COMT anti ⁇ en ⁇ e con ⁇ truct wa ⁇ prepared a ⁇ in Example 1 but using a different fragment of the pCOMT sequence.
  • a 0.6 kb fragment of the alfalfa pCOMT sequence (from the 5' end to the B' site, marked with a thick arrow in FIGURE 5) was u ⁇ ed.
  • the relevant re ⁇ triction enzyme ⁇ ite ⁇ on the pCOMT sequence are depicted in FIGURE 7b.
  • the portion of the pCOMT sequence used for the alfalfa construct is highlighted in FIGURE 7b and is essentially the ⁇ equence located between the BamHI and the leftmost BelI restriction enzyme sites depicted.
  • Insertion of thi ⁇ portion of the sequence in inverted orientation into vector pRTL2 ensured transcription of the complementary, antisen ⁇ e ⁇ trand. Thirty four tran ⁇ genic alfalfa plants were generated which all exhibited NPT II activity. Control plants were taken through the full regeneration protocol.
  • COMT activity was determined as described for tobacco. The result ⁇ are depicted in FIGURE 6. COMT activity in anti ⁇ en ⁇ e plant ⁇ averaged approximately 50% of that of the control. The values for the control are the average and spread of values (as indicated by the bars) from four independent control plants. Total lignin estimation
  • the thioglycolic acid method was used for lignin estimation as described for tobacco. Result ⁇ of COMT activity assay and lignin estimation for transgenic alfalfa are shown in FIGURE 6. These data are for the upper parts of whole seedlings (including leaves) . Lignin content averaged approximately 80% of the control. No direct correlation between lignin content and COMT activity was observed. However, 14 of the 34 tran ⁇ genic plants exhibited both lower COMT activity (35-80%) and lignin content (72-94%) as compared with controls.
  • MOLECULE TYPE cDNA to mRNA
  • HYPOTHETICAL NO
  • ANTI-SENSE NO
  • ORIGINAL SOURCE
  • GCT CAT AAT CCT GGT GGG AAA GAG AGA ACA CAA AAA GAG TTT GAG GAT 1065 Ala His Asn Pro Gly Gly Lys Glu Arg Thr Gin Lys Glu Phe Glu Asp 325 330 335

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Abstract

La présente invetnion se rapporte à un procédé et un réactif de réduction de la teneur en lignine dans des plantes. L'invention implique spécifiquement l'incorporation d'un gène antisens pour la 3-O-méthyltransférase de l'acide caféique dans le génome de plantes.
PCT/US1994/003356 1993-04-02 1994-03-31 Procede de reduction de la teneur en lignine dans des plantes WO1994023044A1 (fr)

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US08/045,263 1993-04-02

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Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
US5973228A (en) * 1997-07-24 1999-10-26 University Of British Columbia Coniferin beta-glucosidase cDNA for modifying lignin content in plants
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
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
WO2006012594A2 (fr) * 2004-07-24 2006-02-02 The Samuel Roberts Noble Foundation, Inc. Modification de la biosynthese de la lignine
US7087426B2 (en) 1996-09-11 2006-08-08 Agrigenesis Biosciences Ltd. Materials and methods for the modification of plant lignin content
EP1826265A2 (fr) 1998-10-09 2007-08-29 Arborgen, Llc Matériaux et procédés pour la modification de la teneur en lignine d'une plante
US7365186B2 (en) 2002-11-22 2008-04-29 Arborgen, Llc Vascular-preferred promoter sequences and uses thereof
US7507875B2 (en) 2003-06-06 2009-03-24 Arborgen, Llc Transcription factors
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
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
US9845478B2 (en) 2010-09-14 2017-12-19 Board Of Trustees Of Michigan State University Compositions and methods for xylem-specific expression in plant cells
CN110016479A (zh) * 2019-05-17 2019-07-16 河南省农业科学院畜牧兽医研究所 一个紫花苜蓿MsGPF基因

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GOWRI, G., ET AL.: "Stress responses in alfalfa (Medicago sativa L.) . X. Molecualr cloning and expression of S-adenosyl-L-methionine:caffeic acid 3-O-methyltransferase, a key enzyme of lignin biosynthesis", PLANT PHYSIOLOGY, vol. 97, 1991, pages 7 - 14 *
NI, W., ET AL.: "Modification of lignin biosynthesis by genetic manipulation of caffeic acid O-methyltransferase", J. CELL. BIOCHEM. SUPPL., KEYSTONE SYMPOSIUM ON CROP IMPROVEMENT VIA BIOTECHNOLOGY : AN INTERNATIONAL PERSPECTIVE, HELD APRIL 10-16, 1992., vol. 16F, 1992, pages 219 *
PODILA, G.K., ET AL.: "Antisense expression of an aspen O-methyltransferase construct in transgenic tobacco via Agrobacterium", PLANT PHYSIOLOGY SUPPLEMENT, vol. 99, no. 1, May 1992 (1992-05-01), pages 19 *
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Cited By (36)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
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
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
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
US6204434B1 (en) 1996-09-11 2001-03-20 Genesis Research & Development Corporation Limited Materials and methods for the modification of plant lignin content
US7087426B2 (en) 1996-09-11 2006-08-08 Agrigenesis Biosciences 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
US6410718B1 (en) 1996-09-11 2002-06-25 Genesis Research & Development Corporation Ltd. Materials and methods for the modification of plant lignin content
US5973228A (en) * 1997-07-24 1999-10-26 University Of British Columbia Coniferin beta-glucosidase cDNA for modifying lignin content in plants
EP1826265A2 (fr) 1998-10-09 2007-08-29 Arborgen, Llc Matériaux et procédés pour la modification de la teneur en lignine d'une plante
US6441272B1 (en) 1998-12-02 2002-08-27 The University Of Georgia Research Foundation, Inc. Modification of lignin content and composition in plants
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
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
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
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
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
US7122718B2 (en) 2000-07-28 2006-10-17 Dairy Australia Limited Modification of plant resistance to diseases and/or pests
US7365186B2 (en) 2002-11-22 2008-04-29 Arborgen, Llc Vascular-preferred promoter sequences and uses thereof
US7442786B2 (en) 2002-11-22 2008-10-28 Arborgen, Llc Vascular-preferred promoters
US8389806B2 (en) 2002-11-22 2013-03-05 Arborgen Inc. Vascular-preferred promoters
US7999149B2 (en) 2002-11-22 2011-08-16 Arborgen, Llc Vascular-preferred promoter sequences and uses thereof
US8110723B2 (en) 2003-06-06 2012-02-07 Arborgen Inc. Transcription factors
US7507875B2 (en) 2003-06-06 2009-03-24 Arborgen, Llc Transcription factors
US8962947B2 (en) 2003-06-06 2015-02-24 Arborgen Inc. Transcription factors
US8679844B2 (en) 2004-04-20 2014-03-25 Syngenta Participations Ag MADS gene regulatory sequences for expressing gene products in plant reproductive tissue
US8129588B2 (en) 2004-04-20 2012-03-06 Syngenta Participations Ag Regulatory sequences for expressing gene products in plant reproductive tissue
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
US9238818B2 (en) 2004-04-20 2016-01-19 Syngenta Participations Ag Methods and genetic constructs for modification of lignin composition of corn cobs
WO2006012594A2 (fr) * 2004-07-24 2006-02-02 The Samuel Roberts Noble Foundation, Inc. Modification de la biosynthese de la lignine
WO2006012594A3 (fr) * 2004-07-24 2006-09-28 Samuel Roberts Noble Found Inc Modification de la biosynthese de la lignine
US7663023B2 (en) 2004-07-24 2010-02-16 The Samuel Roberts Noble Foundation Modification of lignin biosynthesis
US9845478B2 (en) 2010-09-14 2017-12-19 Board Of Trustees Of Michigan State University Compositions and methods for xylem-specific expression in plant cells
CN110016479A (zh) * 2019-05-17 2019-07-16 河南省农业科学院畜牧兽医研究所 一个紫花苜蓿MsGPF基因
CN110016479B (zh) * 2019-05-17 2022-08-26 河南省农业科学院畜牧兽医研究所 一个紫花苜蓿MsGPF基因

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