WO1999066785A1 - Plantes cerealieres transgeniques tolerantes aux stress hydrique et salin - Google Patents

Plantes cerealieres transgeniques tolerantes aux stress hydrique et salin Download PDF

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WO1999066785A1
WO1999066785A1 PCT/US1999/014336 US9914336W WO9966785A1 WO 1999066785 A1 WO1999066785 A1 WO 1999066785A1 US 9914336 W US9914336 W US 9914336W WO 9966785 A1 WO9966785 A1 WO 9966785A1
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cereal plant
transgenic
protoplast
stress
nucleic acid
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PCT/US1999/014336
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Ray J. Wu
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Cornell Research Foundation, Inc.
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Priority to AU47164/99A priority Critical patent/AU4716499A/en
Priority to CA002335522A priority patent/CA2335522A1/fr
Publication of WO1999066785A1 publication Critical patent/WO1999066785A1/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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0008—Oxidoreductases (1.) acting on the aldehyde or oxo group of donors (1.2)
    • 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
    • C12N15/8251—Amino acid content, e.g. synthetic storage proteins, altering amino acid 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
    • 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
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance
    • 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.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)

Definitions

  • the present invention relates to transgenic cereal plants which are transformed with a nucleic acid encoding an enzyme for proline biosynthesis that confers water stress or salt stress tolerance to the plant and a method of increasing or conferring water stress or salt stress tolerance to a cereal plant.
  • the present invention is directed to overcoming the above-noted deficiencies in the prior art.
  • the present invention relates to a transgenic cereal plant transformed with a nucleic acid encoding an enzyme for proline biosynthesis that confers water stress or salt stress tolerance to the plant.
  • the present invention also relates to a cereal plant cell or protoplast transformed with a nucleic acid encoding an enzyme for proline biosynthesis that confers water stress or salt stress tolerance on a cereal plant regenerated from said cereal plant cell or protoplast.
  • Another aspect of the present invention is a method of conferring water stress or salt stress tolerance to a cereal plant including transforming a cereal plant cell or protoplast with a nucleic acid encoding an enzyme for proline biosynthesis.
  • the present invention also relates to a method of increasing tolerance of a cereal plant to water stress or salt stress conditions, the method including increasing levels of an enzyme for proline biosynthesis in the cereal plant.
  • the present invention allows the production of cereal plants with increased tolerance to water stress (drought) and salt stress.
  • an enzyme for proline biosynthesis can be used as a molecular tool for genetic crop improvement by conferring stress tolerance.
  • the present invention relates to a transgenic cereal plant transformed with a nucleic acid encoding an enzyme for proline biosynthesis that confers water stress or salt stress tolerance to the plant.
  • Suitable nucleic acids encoding an enzyme for proline biosynthesis include the P5CS gene of mothbean and a feedback-inhibition insensitive mutant. P5CS- 29A. of the P5CS gene.
  • the sequence of the P5CS gene can be found in Kishor et al.. "Overexpression of ⁇ -pyrroline-5-carboxylate Synthetase Increases Proline Production and Confers Osmotolerance in Transgenic Plants. '' Plant Phvsiol.. 108: 1387-1394 (1995).
  • Cereal which can be transformed in accordance with the subject invention are members of the family Gramineae (also known as Poaceae). and include rice (genus Oryza). wheat, maize (corn), barley, oat. sorghum, and millet.
  • the cereal is rice, wheat, or corn, and most preferably the cereal is rice.
  • Many species of cereals can be transformed, and within each species the numerous subspecies and varieties can be transformed.
  • Within the rice species is subspecies Indica rice (Ory ⁇ a saliva ssp. Indica). which includes the varieties IR36. IR64. IR72. Pokkali. Nona Bokra. KDML105. Suponburi 60. Suponburi 90. Basmati 385.
  • plant cells suitable for transformation include immature embryos, calli. suspension cells, and protoplasts. It is particularly preferred to use suspension cells and immature embryos.
  • nucleic acid which could be RNA or DNA and which is preferably cDNA.
  • the nucleic acid can be biologically isolated or synthetic.
  • a key enzyme for proline biosynthesis ⁇ '-pyrroline-5-carboxylate synthase (P5CS).
  • P5CS ⁇ '-pyrroline-5-carboxylate synthase
  • P5CS- ⁇ 29A other genes encoding an enzyme for proline biosynthesis, including a feedback-inhibition insensitive mutant of the P5CS gene.
  • P5CS- ⁇ 29A can also be utilized.
  • Transformation of plant cells can be accomplished by using a plasmid.
  • the plasmid is used to introduce the nucleic acid encoding an enzyme for proline biosynthesis into the plant cell.
  • a plasmid preferably includes DNA encoding an enzyme for proline biosynthesis inserted into a unique restriction endonuclease cleavage site.
  • Heterologous DNA refers to DNA not normally present in the particular host cell transformed by the plasmid.
  • DNA is inserted into the vector using standard cloning procedures readily known in the art. This generally involves the use of restriction enzymes and DNA ligases. as described by Sambrook et al.. Molecular Cloning: A Laboratory Man ⁇ al. 2d edition. Cold Spring Harbor Laboratory Press. Cold Spring Harbor.
  • the resulting plasmid which includes nucleic acid encoding an enzyme for proline biosynthesis can then be used to transform a host cell, such as an Agrobacte ⁇ um and/or a plant cell.
  • a host cell such as an Agrobacte ⁇ um and/or a plant cell.
  • the plasmid preferably also includes a selectable marker for plant transformation.
  • plant selectable markers include the hygromycin phosphotransferase (hpt) gene, the phosphinothricin acetyl transferase gene (bar), the 5-enolpyruvylshikimate-3-phosphate synthase gene (EPSPS). neomycin 3 " -O- phosphotransferase gene (npt II). or acetolactate synthase gene (ALS). Information on these selectable markers can be found in Bowen. "Markers for Plant Gene Transfer " in Transgenic Plants. Kung et al.. Eds.. Vol. 1. pp. 89-123. Academic Press. NY ( 1993). which is hereby incorporated by reference.
  • the plasmid preferably also includes suitable promoters for expression of the nucleic acid encoding an enzyme for proline biosynthesis and for expression of the marker gene.
  • the cauliflower mosaic virus 35S promoter is commonly used for plant transformation, as well as the rice actin 1 gene promoter.
  • the nucleic acid encoding an enzyme for proline biosynthesis is under the control of the constitutive rice actin 1 gene promoter and the marker gene (bar) is under control of the cauliflower mosaic virus 35S promoter.
  • Other promoters useful for plant transformation w ith an enzyme for proline biosynthesis include those from the genes encoding ubiquitin and proteinase inhibitor II (PINII ).
  • pJS l 12 has been deposited pursuant to. and in satisfaction of. the requirements of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure, with the American Type Culture Collection (ATCC). 10801 University Boulevard. Manassas. VA 201 10-2209. under ATCC Accession No. on June 17. 1999.
  • ATCC American Type Culture Collection
  • the plasmid also preferably includes a nucleic acid molecule encoding a 3 ' terminator such as that from the 3 ' non-coding region of genes encoding a proteinase inhibitor, actin. or nopaline synthase (nos).
  • plasmids for use in the subject invention can be constructed.
  • genes encoding an enzyme for proline biosx nthesis other than the P5CS gene of mothbean could be ligated into plasmid .IS 109 after use of restriction enzymes to remove the P5CS gene.
  • Other promoters could replace the actin 1 gene promoter present in plasmid JS102.
  • other plasmids in general containing genes encoding an enzyme for proline biosynthesis under the control of a suitable promoter, with suitable selectable markers can be readily constructed using techniques well known in the art. Having identified the plasmid.
  • one technique of transforming cereal plant cells with a gene which encodes for an enzyme for proline biosynthesis is by contacting the plant cell with an inoculum of an Agrobacterium bacteria transformed with the plasmid comprising the gene that encodes for the enzyme for proline biosynthesis.
  • this procedure involves inoculating the plant cells with a suspension of the transformed bacteria and incubating the cells for 48 to 72 hours on regeneration medium without antibiotics at 25-28°C.
  • Bacteria from the genus Agrobacterium can be utilized to transform plant cells. Suitable species include Agrobacterium lumefacien and Agrobacterium rhizogenes. Agrobacterium tumefaciens (e.g.. strains LBA4404 or EHA105) is particularly useful due to its well-known ability to transform plants.
  • the bacteria In inoculating the cells of cereal plants with Agrobacterium according to the subject invention, the bacteria must be transformed with a vector which includes a gene encoding for an enzyme for proline biosynthesis.
  • T-DNA plasmid vector pBI121 which contains a low -copy RK2 origin of replication, the neomycin phosphotransferase (nptll) marker gene with a nopaline synthase (NOS) promoter and a NOS 3 " polyadenylation signal.
  • nptll neomycin phosphotransferase
  • NOS nopaline synthase
  • GUS beta-glucuronidase
  • Agrobacterium spp. are transformed with a plasmid by direct uptake of plasmid DNA after chemical and heat treatment, as described by Holsters et al. "Transfection and Transformation of Agrobacterium lumefaciens. " Mol. Gen. Genet.. 163: 181 -187 (1978). which is hereby incorporated by reference: by direct uptake of plasmid DNA after electroporation. as described by Shen et al.. "Efficient Transformation of Agrobacterium spp. by High Voltage Electroporation. " Nucleic Acids Research. 17:8385 ( 1989).
  • Another method for introduction of a plasmid containing nucleic acid encoding an enzyme for proline biosynthesis into a plant cell is by transformation of the plant cell nucleus, such as by particle bombardment.
  • particle bombardment also known as biolistic transformation
  • this procedure involves propelling inert or biologically active particles at the cells under conditions effective to penetrate the outer surface of the cell and to be incorporated within the interior thereof.
  • the plasmid can be introduced into the cell by coating the particles with the plasmid containing the heterologous DNA.
  • the target cell can be surrounded by the plasmid so that the plasmid is carried into the cell by the wake of the particle.
  • Biologically active particles e.g.. dried bacterial cells containing the plasmid and heterologous DNA
  • a further method for introduction of the plasmid into a plant cell is by transformation of plant cell protoplasts (stable or transient). Plant protoplasts are enclosed only by a plasma membrane and will therefore take up macromolecules like heterologous DNA. These engineered protoplasts can be capable of regenerating whole plants.
  • Suitable methods for introducing heterologous DNA into plant cell protoplasts include electroporation and polyethylene glycol (PEG) transformation.
  • electroporation is a transformation method in which. generall ⁇ . a high concentration of plasmid DNA (containing heterologous DNA) is added to a suspension of host cell protoplasts and the mixture shocked with an electrical field of 200 to 600 V/cm. Following electroporation. transformed cells are identified by growth on appropriate medium containing a selective agent.
  • transformation encompasses stable transformation in which the plasmid is integrated into the plant chromosomes.
  • rice has been transformed using biolistic transformation.
  • Other methods of transformation have also been used to successfully transform rice plants, including the protoplast method (for a review, see Cao et al.. "Regeneration of Herbicide Resistant Transgenic Rice Plants Following Microprojectile- Mediated Transformation of Suspension Culture Cells. "* Plant Cell Rep.. 1 1 :586-591 (1992). which is hereby incorporated by reference), and the Agrobacterium method (Hiei et al.. "Efficient Transformation of Rice (On -a saliva L.) Mediated by Agrobacterium and Sequence Analysis of the Boundaries of the T-DNA. " The Plant Journal. 6:271 -282 ( 1994). which is hereby incorporated by reference).
  • Biolistic transformation has also been used to successfully transform maize (for a review, see Mackey et al.. ' ransgenic Maize. " In Transgenic Plants. Kung et al.. Eds... ⁇ ol. 2. pp. 21 -33 (1993). which is hereb> incorporated by reference) and wheat (see U.S. Patent No. 5.405.765 to Vasil et al.. which is hereby' incorporated by reference).
  • a cereal plant cell or protoplast is transformed in accordance with the present invention, it is regenerated to form a transgenic cereal plant.
  • regeneration is accomplished by culturing transformed cells or protoplasts on medium containing the appropriate growth regulators and nutrients to allow for the initiation of shoot meristems. Appropriate antibiotics are added to the regeneration medium to inhibit the growth of Agrobacterium or other contaminants and to select for the development of transformed cells or protoplasts. Following shoot initiation, shoots are allowed to develop in tissue culture and are screened for marker gene activity.
  • the cereal plant cell to be transformed can be in vitro or in vivo. i.e. the cereal plant cell can be located in a cereal plant.
  • the invention also provides seed produced by the transgenic cereal plant.
  • the invention is also directed to seed, which upon germination, produces the transgenic cereal plant.
  • transgenic cereal plants transformed with fragments of the nucleic acids encoding an enzyme for proline biosynthesis of the present invention.
  • Suitable fragments capable of conferring water stress or salt stress tolerance to cereal plants can be constructed by using appropriate restriction sites.
  • a fragment refers to a continuous portion of the encoding molecule for an enzyme for proline biosynthesis that is less than the entire molecule.
  • Non-essential nucleotides could be placed at the 5 ' and/or 3 " ends of the fragments (or the full length molecules encoding an enzyme for proline biosynthesis) without affecting the functional properties of the fragment or molecule (i.e. in increasing water stress or salt stress tolerance).
  • the nucleotides encoding an enzyme for proline biosynthesis may be conjugated to a signal (or leader) sequence at the N- terminal end (for example) of the enzyme for proline biosynthesis which co- translationally or post-translationally directs transfer of the enzyme for proline biosynthesis.
  • the nucleotide sequence may also be altered so that the encoded enzyme is conjugated to a linker or other sequence for ease of synthesis, purification, or identification of the enzyme.
  • the present invention also relates to a cereal plant cell or protoplast transformed with a nucleic acid encoding an enzy me for proline biosynthesis that confers water stress or salt stress tolerance on a cereal plant regenerated from said cereal plant cell or protoplast. Once transformation has occurred, the cereal plant cell or protoplast can be regenerated to form a transgenic cereal plant.
  • the nucleic acid encoding an enzyme for proline biosynthesis is controlled by a strong promoter to effect maximum expression of an enzyme for proline biosynthesis, or by a stress-induced promoter to effect induction of the promoter in response to stress conditions.
  • the transgenic cereal plant cell or protoplast or plant is transformed with the nucleic acid encoding the promoter, such as the rice actin 1 gene promoter, by providing a plasmid hich includes DNA encoding an enzyme for proline biosynthesis and the promoter.
  • the transgenic cereal plant cell or protoplast or plant can also be transformed with a nucleic acid encoding a selectable marker, such as the bar gene, to allow for detection of transformants. and with a nucleic acid encoding the cauliflower mosaic virus 35S promoter to control expression of the bar gene.
  • selectable markers include genes encoding EPSPS. nptll. or ALS.
  • Other promoters include those from genes encoding actin 1. ubiquitin. and PINII.
  • These additional nucleic acid sequences can also be provided by the plasmid encoding the enzyme for proline biosynthesis and its promoter. Where appropriate, the various nucleic acids could also be provided by transformation with multiple plasmids.
  • nucleotide sequence referred to herein encodes an enzyme for proline biosynthesis
  • nucleotide identity to previously sequenced enzymes for proline biosynthesis is not required.
  • various nucleotide substitutions are possible which are silent mutations (i.e. the amino acid encoded by the particular codon does not change). It is also possible to substitute a nucleotide which alters the amino acid encoded by a particular codon. where the amino acid substituted is a conservative substitution (i.e. amino acid "homology " is conserved).
  • the present invention is also directed to a transgenic cereal plant regenerated from the transgenic cereal plant cells or protoplasts, as well as to seed produced by the transgenic cereal plants.
  • Another aspect of the present invention is a method of conferring water stress or salt stress tolerance to a cereal plant including transforming a cereal plant cell or protoplast with a nucleic acid encoding an enzyme for proline biosynthesis.
  • the method further includes regenerating the transformed cereal plant cell or protoplast to form a transgenic cereal plant.
  • the present invention also includes seed produced by the transgenic cereal plant.
  • the present invention also relates to a method of increasing tolerance of a cereal plant to water stress or salt stress conditions, the method including increasing levels of an enzyme for proline biosynthesis in the cereal plant.
  • the plasmid is designated p.JS102. p.ISl 07. or pJS l 12 (See Examples 1 and 2).
  • Plasmid C 'onslruclion p.IS107 was constructed by isolating a 2.4 kb Sail fragment containing mothbean ( Vigna aconitifolia L.) P5CS cDNA from the plasmid pUbiP5CS (Hu et al.. "A Bifunctional Enzyme ( ⁇ '-pyrroline-5-carboxylate synthetase) Catalyzes the First Two Steps in Pro Biosynthesis in Plants. " Proc. Natl. Acad. Sci. USA, 89:9354-9358 ( 1992). which is hereby incorporated by reference).
  • p.TS107 also contains the bar cassette, which was used for selection of transgenic calli and plants in the presence of the herbicide. Bialaphos.
  • pJSl 07 was introduced into suspension culture cells by the biolistic method.
  • the cells were cultured and selected in KPR medium (Cao et al.. "Assessment Of Rice Genetic Transformation Techniques. In Rice Biotechnology. Toenniessen et al.. Eds... CAB International. Oxon. UK. pp. 175-198 (1991 ). which is hereby incorporated by reference) containing 8 mg per liter Bialaphos.
  • the resistant calli were transferred to MS regeneration medium to regenerate into plants. Plants regenerated from the same resistant callus were regarded as clones of the same line. Regenerated plants were transferred into soil and grown in the greenhouse (32°C day/22°C night, with supplemental photoperiod of 10 hours).
  • Plasmid pJS 107 ( ABRC 1 /Act- 100 promoter/H va22 intron/PiCS' cDNA/Pin2 37/35S promoter/ ⁇ w Nos 3 " ) was introduced into rice suspension cells using the biolistic-mediated transformation method. Regeneration and Analysis of Transgenic Plants
  • transgenic rice plants produced an increased level of the P5CS enzyme activity as well as proline content (measured by using a colorimetric method) in leaves.
  • plasmids Three plasmids were constructed. The components of these plasmids are: p.IS102 (with a constitutive promoter): Rice actin 1 promoterAPJCS' cDNA/Pin 2 37/35S promoter/( /Nos 3 " ; pJSl 12 (with a stress-inducible promoter): ABRC4/Actl-100 promoter///vi/22 intron/ ⁇ )GS' cDNA/Pin2 37/35S promoterA Nos 3 * ; and pJSl 10 (with a constitutive promoter and all components as in pJS l 12. except that a uidA reporter gene is used in place of the P5CS cDNA in pJSl 12). Trunsformation of Rice Calli with a Mothbean P5CS cDNA
  • Refined and sterilized field soil was used to gro the rice plants in the greenhouse.
  • R2 seeds were germinated in 1/2 MS medium for 7 days, and the 7-day-old seedlings were transplanted into soil in small pots (8x8 inches) with holes in the bottom (4 to 6 plants per pot).
  • the pots were kept in flat-bottomed trays containing water.
  • the seedlings were grown for an additional 2 weeks, and within the third week, they were tested for Basta resistance. Two Basta-resistant plants with the same plant height per pot were selected for stress treatments. Stress treatments were carried out as follows.
  • the idA plants were chosen as more suitable control plants for the following experiment because they also contained bar and the same promoter cassette as the J -transgenic plants.
  • Fresh shoot and root weights are in mg/plant. Means ⁇ SE represents the averages of 6 plants ( Wt). Values in parentheses are the percentages of/5c.y-transgenic plants compared to control plants (L3). represented by 100. The spread of data within each set of 6 plants was rather small. For example, the actual values for the fresh shoot yvt of six JS110 (L3) plants in the yvater-stress experiment (top half of table) were: 280.282.288, 315.320 and 325; the actual values for the fresh shoot yvt of six JS112 (L5) plants were: 840, 845.860.1025.1045 and 1050.

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Abstract

La présente invention se rapporte à une plante céréalière transgénique et à une cellule de plante céréalière ou protoplaste transformée au moyen d'un acide nucléique codant une enzyme pour réaliser une biosynthèse de la proline pouvant conférer à la plante la tolérance aux stress hydrique et salin. Dans un autre aspect de l'invention, on décrit un procédé pouvant être mis en oeuvre pour conférer à une plante céréalière la tolérance aux stress hydrique et salin, notamment par transformation d'une cellule de plante céréalière ou protoplaste au moyen d'un acide nucléique codant une enzyme pour réaliser une biosynthèse de la proline. L'invention se rapporte en outre à un procédé qui permet d'accroître la tolérance d'une plante céréalière à des conditions de stress hydrique ou salin, le procédé consistant à augmenter les niveaux d'une enzyme pour réaliser la biosynthèse de la proline dans la plante céréalière. Dans un aspect encore différent de l'invention, on décrit une plante céréalière transgénique transformée au moyen d'un plasmide qui confère à la plante céréalière la tolérance aux stress hydrique et salin.
PCT/US1999/014336 1998-06-24 1999-06-24 Plantes cerealieres transgeniques tolerantes aux stress hydrique et salin WO1999066785A1 (fr)

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CA002335522A CA2335522A1 (fr) 1998-06-24 1999-06-24 Plantes cerealieres transgeniques tolerantes aux stress hydrique et salin

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Cited By (6)

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EP1252292A2 (fr) * 2000-01-24 2002-10-30 RAMOT UNIVERSITY AUTHORITY FOR APPLIED RESEARCH & INDUSTRIAL DEVELOPMENT LTD. Plantes tolerant le stress de l'environnement, procedes d'elaboration pour ce type de plante et sequence polynucleotidique utilisee a cet effet
GB2376236A (en) * 2001-06-08 2002-12-11 Hitachi Ltd Stress tolerant transgenic grass plants with altered proline biosynthesis
WO2012099528A1 (fr) * 2011-01-18 2012-07-26 Swetree Technologies Ab Plantes résistant à la sécheresse et leur procédé de production au moyen de régulateurs transcriptionnels
RU2487166C2 (ru) * 2005-10-24 2013-07-10 Эвоген Лтд Изолированные полипептиды, кодирующие их полинуклеотиды, экспрессирующие их трансгенные растения и способы их использования
US8889949B2 (en) 2002-06-20 2014-11-18 Cornell Research Foundation, Inc. Method for increasing resistance of monocot plants against abiotic stresses, TPSP fusion enzyme gene constructs, and transformants
CN112514793A (zh) * 2020-12-21 2021-03-19 扬州大学 一种耐盐性水稻品种的筛选方法

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EP1252292A4 (fr) * 2000-01-24 2004-11-17 Univ Ramot Plantes tolerant le stress de l'environnement, procedes d'elaboration pour ce type de plante et sequence polynucleotidique utilisee a cet effet
US7385106B2 (en) 2000-01-24 2008-06-10 Ramot At Tel Aviv University Ltd. Plants tolerant of environmental stress conditions, methods of generating same and novel polynucleotide sequence utilized thereby
GB2376236A (en) * 2001-06-08 2002-12-11 Hitachi Ltd Stress tolerant transgenic grass plants with altered proline biosynthesis
GB2376236B (en) * 2001-06-08 2003-08-27 Hitachi Ltd Transgenic rice plant and its family with environmental stress resistant by proline accumulation of high level and its production
US8889949B2 (en) 2002-06-20 2014-11-18 Cornell Research Foundation, Inc. Method for increasing resistance of monocot plants against abiotic stresses, TPSP fusion enzyme gene constructs, and transformants
RU2487166C2 (ru) * 2005-10-24 2013-07-10 Эвоген Лтд Изолированные полипептиды, кодирующие их полинуклеотиды, экспрессирующие их трансгенные растения и способы их использования
WO2012099528A1 (fr) * 2011-01-18 2012-07-26 Swetree Technologies Ab Plantes résistant à la sécheresse et leur procédé de production au moyen de régulateurs transcriptionnels
CN112514793A (zh) * 2020-12-21 2021-03-19 扬州大学 一种耐盐性水稻品种的筛选方法

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