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WO1994028147A1 - Sequence d'adn comprenant au moins deux genes de proteines d'enveloppe - Google Patents

Sequence d'adn comprenant au moins deux genes de proteines d'enveloppe Download PDF

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Publication number
WO1994028147A1
WO1994028147A1 PCT/EP1994/001786 EP9401786W WO9428147A1 WO 1994028147 A1 WO1994028147 A1 WO 1994028147A1 EP 9401786 W EP9401786 W EP 9401786W WO 9428147 A1 WO9428147 A1 WO 9428147A1
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WO
WIPO (PCT)
Prior art keywords
dna sequence
coat protein
proteins
sequence
virus
Prior art date
Application number
PCT/EP1994/001786
Other languages
English (en)
Inventor
Kirsten Bojsen
Janne Brunstedt
Original Assignee
Sandoz Ltd.
Sandoz-Patent-Gmbh
Sandoz-Erfindungen Verwaltungsgesellschaft M.B.H.
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 Sandoz Ltd., Sandoz-Patent-Gmbh, Sandoz-Erfindungen Verwaltungsgesellschaft M.B.H. filed Critical Sandoz Ltd.
Priority to AU71226/94A priority Critical patent/AU7122694A/en
Publication of WO1994028147A1 publication Critical patent/WO1994028147A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • 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
    • C12N15/8279—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 biotic stress resistance, pathogen resistance, disease resistance
    • C12N15/8283—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 biotic stress resistance, pathogen resistance, disease resistance for virus 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
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/00022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • DNA sequence comprisine at least two coat protein genes
  • the present invention relates to transformed plants, particularly sugar beet.
  • a recombinant DNA sequence comprising nucleotide sequences encoding the coat proteins of at least one luteovirus and at least one closterovirus, or a recombinant sequence comprising nucleotide sequences encoding at least a part of both proteins.
  • the recombinant sequence may additionally comprise a nucleotide sequence encoding a coat protein, or a part thereof, of a furo-virus, such as beet necrotic yellow vein virus (BNYVV).
  • part is meant a peptide encoded by the sequence depicted in SEQ ID Nos 1, 2 or 3, and having at least 10 amino acids. More preferably the peptide has at least 15 amino acids, and still more preferably the peptide has at least 20 amino acids.
  • the invention also includes a recombinant DNA sequence according to the invention, which is modified in that codons which are preferred by the organism into which the recombinant DNA is to be inserted are used so that expression of the thus modified DNA in the said organism yields substantially similar proteins to those obtained by expression of the unmodified recombinant DNA in the organism in which the protein-encoding components of the recombinant DNA are found endogenously.
  • the closterovirus is beet yellows virus (BYV), and the luteovirus is beet western yellows virus (BWYV).
  • Transcription of all the coat protein genes may be under control of a single promoter. It is preferred, however, that transcription of each coat protein gene, or part thereof, is under control of a seperate promoter.
  • promoters are known and may be constitutive and or inducible. Suitable promoters include the 35 S promoter, the hsp80 promoter (US Patent Application No 791929), the hsr203 promoter (European Patent Application No. 93102887.2), and promoters regulating expression of octipine genes, rubisco gene(s), and the promoters of genes encoding plant wound-inducible proteins, such as the WUN promoter (European Patent Application No. 89912967.0), and promoters of pathogene sis-related inducible proteins.
  • the transcription of at least the BYV coat protein gene and the BWYV coat protein gene is under control of an enhanced 35 S promoter or the hsr203 promoter or the hsp80 promoter.
  • the recombinant DNA sequence may further comprise other gene regulator sequences, typically down stream of the promoters.
  • Such sequences are known and are preferably positioned between the promoter and the structural gene regulated thereby, and include the omega sequence from tobacco mosaic virus which may function as part of a leader sequence in front of the coat protein genes.
  • the recombinant sequence may further comprise known marker genes and/or at least one gene encoding a known anti-microbial protein.
  • An anti-microbial protein includes a protein (alone or in combination with another material) which is toxic or growth inhibitory under any circumstances to any micro-organism, including bacteria, viruses and particularly fungi.
  • Such anti-microbial proteins include those that exhibit anti-microbial activity upon contact with a micro-organism and those that are anti-microbial as a consequence of assimilation or respiration thereof.
  • PR proteins homologous pathogenesis-related proteins
  • PR proteins homologous pathogenesis-related proteins
  • the PR-2, PR-3 and PR-5 proteins are beta- 1,3 glucanase, chitinases and thaumatin-like proteins respectively. Specific functions have not been assigned to the PR-1 and PR-4 groups of proteins.
  • the PR-4 proteins are similar to C-terminal domains of prohevein and the putative wound-induced WIN proteins of potato, thus lacking the N-terminal hevein domain.
  • “Basic counter-part of the acidic pathogenesis-related 4 group of proteins” thus includes the basic counter part of proteins similar to the C-terminal domains of prohevein and the putative wound-induced WIN proteins of potato.
  • the recombinant DNA sequence according to the present invention may still further comprise at least one gene encoding a protein which is the basic counter-part of the acidic pathogenesis- related 4 group of proteins.
  • the preferred such protein is a chitin-binding WIN protein, preferably similar if not identical to that extractable from barley grain or stressed barley leaves.
  • the recombinant DNA sequence is modified, wherein - at least in the coat protein genes - known plant preferred codons are substituted for viral codons so that transcription of the thus modified DNA in plants and translation of the resulting RNA yields coat proteins having an amino acid sequence identical or substantially similar to the native virus coat proteins.
  • Further preferred modifications to the DNA sequence include removal or deactivation of open reading frames within the viral coat protein genes, particularly removal or deactivation of the internal open reading frame at nucleotides 120-122 in SEQ ID No. 2 within the gene encoding the BWYV coat protein, and alteration of the coat protein gene stop codons.
  • the preferred such alterations include replacement of leaky stop codons (for example TAG) by non-leaky such codons (for example TAA).
  • the invention further includes nucleic acid sequences which are similar to the recombinant DNA sequence according to the invention.
  • a test sequence which is capable of hybridizing to the inventive sequence.
  • the nucleic acid constituting the test sequence preferably has a TM within 20°C of that of the inventive sequence.
  • the TM values of the sequences are preferably within 10°C of each other. More preferably the hybridization is performed under stringent conditions, with either the test or inventive DNA preferably being supported.
  • a denatured test or inventive sequence is preferably first bound to a support and hybridization is effected for a specified period of time at a temperature of between 50 and 65°C in double strength citrate buffered saline containing 0.1%SDS followed by rinsing of the support at the same temperature but with a buffer having a reduced SC concentration.
  • reduced concentration buffers are typically single strength SC containing 0.1%SDS, half strength SC containing 0.1%SDS and one tenth strength SC containing 0.1%SDS.
  • Sequences having the greatest degree of similarity are those the hybridization of which is least affected by washing in buffers of reduced concentration. It is most preferred that the test and inventive sequences are so similar that the hybridization between them is substantially unaffected by washing or incubation in one tenth strength sodium citrate buffer containing 0.1%SDS.
  • the present invention also includes a vector containing any of the above-mentioned DNA sequences, as well as a biological system which includes any of said DNA sequences and which allows expression of the DNA .
  • the biological system may be a micro-organism, such as an Agrobacterium strain, for example, or a plant.
  • the invention also includes plants, particularly sugar beet, transformed with the above disclosed recombinant DNA, and plants transformed with a recombinant DNA sequence including a portion encoding the coat proteins of both the beet yellows virus and the beet western virus, and optionally the coat protein of the beet necrotic yellow vein virus, or similar proteins functionally equivalent thereto in which one or more amino acids have been added, substituted or removed without substantially reducing the capacity of the protein(s) to function as a viral coat protein and/or to elicit a plant mediated ⁇ anti-viral response. It is preferred that such functionally equivalent proteins have an amino acid sequence which is at least 70% similar to the sequence of at least one of the viral coat proteins encoded by the recombinant DNA sequence. It is more preferred that the degree of similarity is at least 80%, still more preferred that the degree of similarity is at least 85% and still more preferred that the degree of similarity is at least 90%.
  • two amino acid sequences with at least 70% similarity to each have at least 70% identical or conservatively replaced amino acid residues in a like position when aligned optimally allowing for up to 5 gaps with the proviso that in respect of the gaps a total not more than 10 amino acid residues are affected.
  • Such plants are made by known methods and include regeneration of plant cells or protoplasts transformed with the recombinant DNA of the invention according to a variety of known methods (Agrobacteriwn Ti and Ri plasmids, electroporation, micro-injection, micro-projectile gun etc).
  • the transformed cells may in suitable cases be regenerated into whole plants in which the nuclear material is stably incorporated into the genome. Both monocot and dicot plants may be obtained in this way.
  • Examples of genetically modified plants according to the present invention include: fruits, including tomatoes, mangoes, peaches, apples, pears, strawberries, bananas, and melons; field crops such as canola, sunflower, tobacco, sugar beet, small grain cereals such as wheat, barley and rice, maize and cotton, and vegetables such as potato, carrot, lettuce, cabbage and onion.
  • fruits including tomatoes, mangoes, peaches, apples, pears, strawberries, bananas, and melons
  • field crops such as canola, sunflower, tobacco, sugar beet, small grain cereals such as wheat, barley and rice, maize and cotton, and vegetables such as potato, carrot, lettuce, cabbage and onion.
  • the preferred plant is sugar beet.
  • the invention also includes the progeny of plants according to the present invention, which progeny expresses the said recombinant DNA sequences, and the seeds of the plants or the progeny thereof.
  • the invention still further includes protein derived from expression of the recombinant DNA according to the present invention, and anti-viral protein produced by expression of the recombinant DNA within plants.
  • the invention still further includes a process for combatting viruses in sugar beet which comprises transforming sugar beet with a recombinant DNA sequence according to the present invention and obtaining transcription of the said DNA. It is particularly preferred that the RNA resulting from the transcription of said recombinant DNA is translated into protein.
  • SEQ ID No. 1 shows the nucleotide sequence of the BYV coat protein gene.
  • SEQ ID No. 2 shows the nucleotide sequence of the BWYV coat protein gene, in which a start codon of an internal open reading frame in the native viral gene has been removed and a leaky stop codon replaced.
  • SEQ ID No. 3 shows the nucleotide sequence of the BNYVV coat protein in which PCR has been used to replace a leaky stop codon (TAG) at bp. 654-656 with the codon TAA.
  • Figure 1 shows the vector pUC19ES35S-35St into which the sequence depicted in SEQ ID No.
  • Figure 2 shows the vector pPS48 into which the sequence depicted in SEQ ID No 2 is cloned as a Hindlll-EcoRI blunt ended fragment in the Smal site, and into which the BNYVV coat protein sequence of SEQ ID No. 3 is cloned as a Hindlll blunt ended fragment.
  • Figure 3 shows the BNYVV coat protein sequence of SEQ ID No. 3 cloned according to
  • Figure 2 and inserted into the plasmid pGNBNYVV.
  • Figure 4 shows the plasmid pGNBWYVBYV comprising the EcoRl fragment resulting from digestion of the vector of Figure 1 and the Hindlll fragment comprising the BWYV coat protein gene resulting from digestion of the vector of Figure 2.
  • Figures 3 and/or 4 the following abbreviations are used:
  • OCSt Octipine synthetase terminator sequence
  • NPTII Neomycin phosphotransf erase encoding gene
  • 35S Cauliflower Mosaic Virus 35S promoter
  • 35St Cauliflower Mosaic Virus 35S terminator sequence
  • E35S Enhanced Cauliflower Mosaic Virus 35S Promoter
  • Omega Omega leader sequence from Tobacco Mosaic Virus
  • BMYV Beet Mild Yellows Virus coat protein
  • BYV Beet Yellows Virus coat protein
  • BNYVV Beet Necrotic Yellow Vein Virus coat protein
  • GUS Beta-glucuronidase encoding protein
  • Figure 5 shows a PCR analysis of sugar beets transgenic for both the BWYV and the BYV coat proteins according to the nucleotide sequences depicted in SEQ ID Nos 1 and 2.
  • the present invention provides, inter alia, a recombinant DNA sequence comprising nucleotide sequences encoding the coat proteins of at least one luteovirus and at least one closterovirus, or a recombinant sequence comprising nucleotide sequences encoding at least a part of both proteins.
  • the recombinant sequence may additionally comprise a nucleotide sequence encoding a coat protein, or a part thereof, of a furo-virus, such as beet necrotic yellow vein virus (BNYVV).
  • BNYVV beet necrotic yellow vein virus
  • the preferred luteovirus is beet western yellows virus (BWYV), and the preferred closterovirus is beet yellows virus (BYV).
  • a vector comprising at least the coat protein genes for the BYV and the BWYV is produced and inserted into a suitable Agrobacteriwn strain according to known methods.
  • the thus transformed Agrobacteriwn is then used to transform plant cells, preferably sugar beet.
  • the transformed cells are then regenerated into plants transgenic for both coat proteins.
  • SEQ ID No. 1 shows the nucleotide sequence of the BYV coat protein gene.
  • the omega sequence from Tobacco Mosaic Virus is inserted as a leader sequence (bp. 13-78) in front of the coat protein gene (between the promoter and the coat protein gene).
  • the start codon is given by bp. 89-91 and the stop codon by bp. 701-703.
  • the BYV coat protein sequence depicted in SEQ ID No. 1 is cloned as a KpnI-BamHI fragment in Pstl-BamHI in pUC19ES35S-35St using a Pstl-Kpnl linker.
  • the EcoRI fragment comprising the BYV coat protein gene resulting from digestion of this vector is inserted into pGNBWYVBYV together with the Hindlll fragment comprising the BWYV coat protein resulting from digestion of the vector depicted in Figure 2 (see Figure 4).
  • SEQ ID No. 2 shows the nucleotide sequence of a modified BWYV coat protein gene.
  • the leaky stop codon (TAG) in the native viral gene sequence is replaced by the codon TAA at bp. 695-697 as can be seen from the sequence given in SEQ ID No. 2, and the start codon of an internal Open Reading Frame (ORF) of the coat protein gene (bp. 120-122. in SEQ ID No. 2) is changed from ATG to ACG in order to prevent translation of this ORF. Both alterations are performed using PCR.
  • SEQ ID No. 2 also shows that the omega sequence from tobacco mosaic virus is inserted as a leader sequence (bp. 7-72) in front of the coat protein gene. The start codon is bp. 89-91.
  • the modified BWYV coat protein gene of SEQ ID No. 2 is cloned as a Hindlll-EcoRI blunt ended fragment in the Smal site of pPS48 (Fig. 2).
  • the Hindlll fragment comprising the BWYV coat protein gene resulting from digestion of the vector of Figure 2 is inserted into the plasmid pGNBWYVBYV together with the EcoRI fragment comprising the BYV coat protein gene resulting from digestion of the vector of Figure 1 (see Figure 4).
  • SEQ LD No. 3 shows the nucleotide sequence of a modified BNYVV coat protein gene, in which PCR is used to replace a leaky stop codon (TAG) in the viral nucleotide sequence at bp. positions 654-656 with TAA.
  • the start codon of the gene depicted in SEQ ID No. 3 is bp. 90-92.
  • the nucleotide sequence of the BNYVV coat protein is further modified, preferably by insertion of the omega sequence from Tobacco Mosaic Virus as a leader sequence (bp- 7-72) in front of the coat protein gene, between it and the promoter.
  • the BNYVV virus coat protein sequence is cloned as a Hindlll blunt-ended fragment in pS48 ( Figure 2) and inserted into pGNBNYVV (see Figure 3).
  • a disarmed Agrobacteriwn twnefaciens strain for example, is transformed with the vector of Figure 4 or a like vector further comprising the BNYVV coat protein gene. Plant cells are then treated with such transformed disarmed Agrobacteriwn, and the thus transformed plant cells are regenerated into whole plants (optionally via callus), in which the new nuclear material is stably incorporated into the genome. Morphologically normal plants which do not contain Agrob ⁇ ctewm-originating oncogenes result from such transformation.
  • the recombinant DNA of the invention may be introduced into plant cells by other known methods, including use of a micro-projectile gun, electroporation, electro- transformation, incubation with plant protoplasts in the presence of polyethylene glycol, and micro-injection etc, and that regeneration of transformed plant cells or protoplasts is carried out according to methods known to the skilled man, including treatment of the cells or protoplasts with cytokines and/or low carbohydrate content media where this is necerney or desirable in order to improve the regeneration frequency.
  • Plants transformed to comprise the DNA sequences of the present invention express the introduced coat protein genes (Figure 5) and exhibit an improved response to challenge with the respective viruses.
  • a particularly beneficial improvement is seen when the transformed plants are challenged with both viruses, either simultaneously by way of a mixed infection, or concomitantly as could occur in nature.
  • Plants subjected to a concomitant infection exhibit a surprisingly improved resistance to the second viral infection in comparison with non- transformed control plants.
  • the DNA sequence may comprise, or further comprise the nucleotide sequence which is complementary to the nucleotide sequence encoding at least one of the virus coat proteins.
  • the DNA sequence may comprise a complementary nucleotide sequence as a substitute for at least one of the genes encoding at least one of the coat proteins of luteo- clostero- or furo- viruses.
  • ORGANISM Beet Yellows Virus
  • ORGANISM Beet Western Yellows Virus
  • ORGANISM Beet Necrotic Yellow Vein Virus
  • GGTTCAAATT ACCATGGACA CCTGTTCAAG GTAGAACCAG TCCACCCGGA CAATAACAAT 660

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Abstract

La présente invention se rapporte à une séquence d'ADN recombinant comprenant des séquences nucléotidiques codant les protéines d'enveloppe d'au moins un lutéovirus et au moins un clostérovirus, ou une séquence recombinante comprenant des séquences nucléotidiques codant au moins une partie des deux protéines. La séquence recombinante peut en outre comprendre une séquence nucléotidique codant une protéine d'enveloppe, ou une partie d'une telle protéine, d'un furo-virus tel que le virus de la rhizomanie de la betterave sucrière.
PCT/EP1994/001786 1993-06-02 1994-06-01 Sequence d'adn comprenant au moins deux genes de proteines d'enveloppe WO1994028147A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU71226/94A AU7122694A (en) 1993-06-02 1994-06-01 Dna sequence comprising at least two coat protein genes

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9311332.2 1993-06-02
GB939311332A GB9311332D0 (en) 1993-06-02 1993-06-02 Improvements in or relating to organic compounds

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WO1994028147A1 true WO1994028147A1 (fr) 1994-12-08

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AU (1) AU7122694A (fr)
GB (1) GB9311332D0 (fr)
IL (1) IL109847A0 (fr)
WO (1) WO1994028147A1 (fr)
ZA (1) ZA943863B (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996021031A1 (fr) * 1994-12-30 1996-07-11 Asgrow Seed Company Plantes transgeniques exprimant des produits de recombinaison d'adn contenant une pluralite de genes destines a leur conferer une resistance virale

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991013159A2 (fr) * 1990-03-02 1991-09-05 Biocem Transformation genetique et regeneration de la betterave sucriere
EP0531273A2 (fr) * 1991-09-03 1993-03-10 Monsanto Company Plantes résistantes aux virus et méthode pour celles-ci

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991013159A2 (fr) * 1990-03-02 1991-09-05 Biocem Transformation genetique et regeneration de la betterave sucriere
EP0531273A2 (fr) * 1991-09-03 1993-03-10 Monsanto Company Plantes résistantes aux virus et méthode pour celles-ci

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
BIOLOGICAL ABSTRACTS, vol. 92, 1991, Philadelphia, PA, US; abstract no. 75334, BRUNSTEDT, J., ET AL.: "Nucleotide sequence of complementary DNA encoding the coat protein of beet yellows virus" *
GIELEN, J.J.L., ET AL.: "Transformation of lettuce cultivars with a BWYV coat protein gene", ABSTRACTS VIITH INTERNATIONAL CONGRESS ON PLANT TISSUE AND CELL CULTURE, AMSTERDAM, JUNE 24-29, 1990., pages 58 *
LAWSON, C., ET AL.: "Engineering resistance to mixed virus infection in a commercial cultivar: resistance to potato virus X and potato virus Y in transgenic russet burbank", BIOTECHNOLOGY, vol. 8, no. 2, February 1990 (1990-02-01), pages 127 - 134 *
VEIDT, I., ET AL.: "Nucleotide sequence of beet western yellows virus RNA", NUCLEIC ACIDS RESEARCH, vol. 16, no. 21, 1988, ARLINGTON, VIRGINIA US, pages 9917 - 9932 *
VEIDT, I., ET AL.: "Synthesis of full-length transcripts of beet western yellows virus RNA: meesenger properties and biological activity in protoplasts", VIROLOGY, vol. 186, 1992, pages 192 - 200 *
VIRUS GENES, vol. 5, no. 3, 1991, pages 267 - 272 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996021031A1 (fr) * 1994-12-30 1996-07-11 Asgrow Seed Company Plantes transgeniques exprimant des produits de recombinaison d'adn contenant une pluralite de genes destines a leur conferer une resistance virale
US6337431B1 (en) 1994-12-30 2002-01-08 Seminis Vegetable Seeds, Inc. Transgenic plants expressing DNA constructs containing a plurality of genes to impart virus resistance
EP1816202A3 (fr) * 1994-12-30 2009-10-28 Seminis Vegetable Seeds, Inc. Plantes transgéniques exprimant les constructions ADN contenant plusieurs gènes pour transmettre la résistance aux virus

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AU7122694A (en) 1994-12-20
ZA943863B (en) 1995-12-04
GB9311332D0 (en) 1993-07-21
IL109847A0 (en) 1994-10-07

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