WO2000001824A2 - Recombinaison genetique de la resistance aux maladies au moyen de la classe de proteines drr206 - Google Patents
Recombinaison genetique de la resistance aux maladies au moyen de la classe de proteines drr206 Download PDFInfo
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- 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
- C12N15/8282—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 fungal resistance
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
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- C12Y302/01014—Chitinase (3.2.1.14)
Definitions
- the present invention relates generally to the field of transgenic plants and expression vectors.
- Plant defense in response to pathogenic attack is accompanied by modification of plant cell structures, synthesis of secondary metabolites and accumulation of specific defense proteins (Staskawicz et al, 1995, Science 268:661-667; Linthorst, 1991 Crit Rev Plant Sci 10: 123-150).
- inducible resistance often results in cell death at the infection site in order to restrict further pathogen growth.
- the area of cell death develops a visible necrotic lesion of a hypersensitive response (HR) (Matthews, 1991 , Plant Virology Academic Press: San Diego).
- HR hypersensitive response
- genes encoding proteins related to defense are expressed (Ward et al, 1991 , Plant Cell 3:1085-1094). Since PR proteins with antimicrobial activity are induced during systemic acquired resistance and hypersensitive responses, these PR proteins may function in protecting plants from pathogenic attack (Lawton et al, 1993).
- the simplest means for genetic engineering of resistance to fungal diseases entails the constitutive expression of one or more defense proteins in transgenic plants.
- Defense proteins with clearly demonstrated antifungal activities have been effective in several cases.
- constitutive expression of bean chitinase protects transgenic tobacco seedlings from Rhizoctonia solani (Brogue et al, 1991 , Science 254:1194-1197).
- transgenic tobacco plants expressing PR1a showed decreased or delayed symptoms both to blue mold (Peronospora tabacina) and black shank (Phytophthora parasitica) (Alexander et al, 1993, Proc Natl Acad Sci USA 90:7327- 7331 ).
- genes other than defense genes have been useful in plant protection.
- Constitutive expression of the fungal elicitor protein ⁇ -crytogein from Phytophthora cryptogea conferred resistance to Phytophthora parasitica in transgenic tobacco.
- constitutive expression of the fungal glucose oxidase gene which generates H 2 0 2 from glucose, resulted in resistance to both Phytophthora infestans, as well as to the bacterium Erwina carotovora (Wu et al, 1995, Plant Cell 7: 1357-1368).
- DRR206 has not previously been used in transgenic plant experiments, although the gene is strongly induced in pea in response to both fungal and bacterial pathogens and elicitors (Riggleman et al, 1985, Plant Mol Bio
- a recombinant expression system capable, when transformed into a plant, of expressing a DNA sequence encoding DRR206 protein, said DRR206 protein having at least 60% identity to amino acids 1-184 of Figure 8, which system comprises control sequences effective in said plant operably linked to said DNA sequence.
- the plant may be Brassica napus.
- control sequences may include a strong constitutive promoter, for example the 35S promoter.
- control sequences may include an inducible promoter.
- the inducible promoter may be responsive to pathogen infection.
- the expression system may include T-DNA for integration of the expression system into a plant genome.
- transgenic plants and plant cells containing the above-described expression system there are provided transgenic plants and plant cells containing the above-described expression system.
- the plants or plant cells may be ⁇ . napus..
- a recombinant expression system capable, when transformed into a plant, of expressing a DNA sequence encoding defensin protein, said defensin protein having at least 60% identity to amino acids 1-72 of Figure 9, which system comprises control sequences effective in said plant operabiy linked to said DNA sequence.
- the plant may be Brassica napus.
- control sequences may include a strong constitutive promoter, for example, the strong constitutive promoter is the 35S promoter.
- the control sequences may include an inducible promoter.
- the inducible promoter may be responsive to pathogen infection.
- the recombinant expression system may include T-DNA for integration of the expression system into a plant genome.
- transgenic plants and plant cells containing the above-described expression system there are provided transgenic plants and plant cells containing the above-described expression system.
- the plants or plant cells may be B. napus.
- a method for producing a plant with improved disease resistance comprising transforming a plant with an expression system comprising a DNA sequence encoding DRR206 protein operabiy linked to control sequences effective in said plant, said DRR206 protein having at least 60% identity to amino acids 1 to 184 of Figure 8; and growing the plant under conditions such that the DRR206 protein is expressed.
- the plant may be resistant to a pathogenic organism selected from the group consisting of Rhizoctonia solani, Leptosphaeria maculans isolate PG3, Leptosphaeria mculans isolate PG4 and Sclerotinia sclerotiorum.
- a method for producing a plant with improved disease resistance comprising transforming a plant with an expression system comprising a DNA sequence encoding defensin protein operabiy linked to control sequences effective in said plant, said defensin protein having at least 60% identity to amino acids 1 to 72 of
- the plant may be resistant to a pathogenic organism selected from the group consisting of Rhizoctonia solani, Leptosphaeria maculans isolate PG3, Leptosphaeria mculans isolate PG4 and Sclerotinia sclerotiorum.
- plants and seeds produced according to the above-described method there are provided plants and seeds produced according to the above-described method.
- a method for producing a transgenic ⁇ . napus plant comprising: providing seeds of a ⁇ . napus strain; providing a recombinant expression system, capable, when transformed into B. napus, of expressing a DNA sequence encoding DRR206 protein, said DRR206 protein having at least 60% identity to amino acids 1 to 184 of Figure 8, which system comprises control sequences effective in ⁇ . napus operabiy linked to said DNA sequence; transfecting Agrobacterium with the recombinant expression system; germinating the ⁇ .
- napus seeds removing cotyledons from the germinated seeds; cocultivating the cotyledons with the transfected Agrobacterium; regenerating shoots and roots from the cotyledons; and growing the transgenic ⁇ . napus plants to maturity.
- a method of providing resistance in ⁇ . napus to a pathogenic organism comprising: providing seeds of a ⁇ . napus strain; providing a recombinant expression system, capable, when transformed into ⁇ . napus, of expressing a DNA sequence encoding DRR206 protein, said DRR206 protein having at least 60% identity to amino acids 1-184 of Figure 8, which system comprises control sequences effective in ⁇ . napus operabiy linked to said DNA sequence; transfecting Agrobacterium with the recombinant expression system; germinating the ⁇ .
- the pathogenic organism may be selected from the group consisting of Rhizoctonia solani, Leptosphaeria maculans isolate PG3, Leptosphaeria mculans isolate PG4 and Sclerotinia sclerotiorum.
- a method of reducing damage to a plant by a pathogenic organism comprising: providing a plant having cells arranged to contain elevated levels of DRR206 protein, said DRR206 protein having at least 60% identity with amino acids 1-184 of Figure 8; and growing the plant such that DRR206 is expressed within the cells of the plant, thereby preventing or inhibiting growth of the pathogenic organism.
- the pathogenic organism may be selected from the group consisting of Rhizoctonia solani, Leptosphaeria maculans isolate PG3, Leptosphaeria maculans isolate PG4 and Sclerotinia sclerotiorum.
- Figure 1 is a diagram of the constitutive defense gene constructs in pBI121.
- Figure 2 is a bar graph summarizing the distribution of copy number in transgenic lines.
- Figure 3 is a bar graph of disease score and mRNA expression in transformed lines.
- Figure 4 summarizes infection phenotypes of transgenic and untransformed B. napus inoculated with Leptosphaeria maculans PG2 (Rimmer and van den Berg, 1992, Can J Plant Pathol 14:56-66).
- Figure 4a-c shows inoculation of cotyledons by pinprick, wherein 4a is untransformed Westar, 4b is defensin transformant GN4-2#15 and 4c is DRR206 transformant GN3-4#22
- Figure 4d shows adult plants inoculated at stems (left to right: untransformed Westar, GN4-2#15 with defensin, GN3-2#22 with DRR206;
- Figure 4e-g shows close-ups of stems shown in Figure 4d;
- Figure 4h-l shows hyphal growth in cotyledon assay 8 days postinoculation;
- Figure 4k shows inoculation of cotyledons by infiltration 8 days postinoculation.
- Figure 5 is an immunoblot of DRR206 proteins in transgenic ⁇ . napus and Fivsar/t/m-inoculated pea.
- Figure 6 is a bar graph showing disease score of DRR206- transformed or untransformed ⁇ . napus inoculated with L. maculans PG3.
- Figure 7 is a bar graph showing disease score of DRR206- transformed or untransformed B. napus inoculated with Sclerotinia sclerotiorum.
- FIG. 8 shows the protein and DNA sequences of DRR206.
- Figure 9 shows the protein and DNA sequences of defensin.
- Table 1 summarizes specific RNA accumulation and disease scores in ⁇ . napus transformed with defense genes.
- Table 2 shows that disease score co-segregates with defense T-DNA insertions in T2 plants.
- Table 3 shows germination rate of pycnidiospores of L. maculans and dry weight of hyphae of L. maculans.
- Table 4 shows disease scores of adult transgenic plant stems inoculated with PG3.
- Table 5 shows disease scores of transgenic plants inoculated with PG4.
- Table 6 shows death rates of transgenic plants inoculated with Rhizoctonia solani.
- Table 7 shows lesion sizes of transgenic plants inoculated with Sclerotinina sclerotiorum.
- canola refers to specific cultivars of Brassica napus and Brassica rapa having low erusic acid and low liolenic acid.
- preventing infection refers generally to providing resistance to disease caused by pathogens, where disease resistance consists of a reduction of symptoms caused by infection, prevention of sporulation or a reduction in damage to the plant by the pathogen. These symptoms may include but are not limited to necrosis, tissue collapse, growth of the pathogen within plant tissues, and decreases in yield and/or quality due to infection.
- mode of pathogenicity refers to the part of a plant infected by a pathogen.
- Expression vectors comprising protein coding sequences for PR10, chitinase, DRR206 and defensin were cloned into a T-DNA-based binary vector under control of a strong constitutive promoter, in this case, the 35S promoter. The vectors were then transformed into B. napus under conditions wherein the T-DNA regions integrated into the ⁇ . napus genome. Transformed plants were screened for cell lines containing single copies of the genes. Cell lines were then assayed for resistance to Leptosphaeria maculans, the blakleg fungus. Results indicate that lines transformed with PR10 or chitinase showed little or no inhibition of infection while lines transformed with DRR206 or defensin did.
- DRR206 is known to be activated in pea in response to a variety of different species of bacteria and fungi, suggesting that DRR206 will confer resistance to a broader range of pathogens. This is supported by the fact that
- DRR206 confers resistance to other pathogens, for example, Sclerotinia sclerotiorum and Rhizoctonia solani. Furthermore, as the DRR206 sequence is not naturally-occurring in B. napus, the presence of the gene in progeny from a cross can be verified using molecular methods rather than infection phenotype. Thus, blackleg resistance can be quickly and easily transferred to any cultivars of B. napus.
- the 35S-constructs were prepared as follows: genomic or cDNA coding sequences for pea PR10 (cDNA) (Drr49) (Culley et al, 1995a, Plant Physiol
- the spore concentration was adjusted to 2x10 7 spores/ml for inoculation.
- Cotyledon inoculation Cotyledons of eight-day plants were wounded with a syringe needle and 10 ⁇ l of the pycnidiospore suspension was dropped onto the lesion. Disease scores were evaluated at 10 days post-inoculation using a disease rating scale based on lesion size, tissue blacking, tissue collapse and presence of pycnidia (Williams and Dehwiche, 1980, Eucarpia "Cruciferae 1979" conference. Wageningen : pp164-170). Cotyledons from Westar control and from transgenic plants with the DRR206 and defensin genes were infiltrated with the spore suspension and the samples were collected post inoculation each day for 8 days.
- Tissue was fixed in glacial acetic acid and absolute ethanol (1 :2, v/v). The fixed solution was changed twice daily for three days. Unstained tissue was then observed under the microscope to observe the morphological characteristics of the blackleg fungus, as described below.
- Bioassays for antifungal activity Uninoculated young leaves of ⁇ . napus cv. Westar or transgenic lines carrying DRR206 or defensin were ground under liquid N 2 and 1 ml of sterile distilled water was added per gram of fresh weight. Insoluble components were removed by centrifugation at 4000 g. The extract was filtered through a 0.2 ⁇ M filter. L. maculans pycnidiospores were added to the extract to a concentration of 1 x 10 6 spores/ml and incubated on a shaker at 120 rpm at room temperature for 72 hours. Germination was quantified in a hemocytometer, using four duplicate grids per treatment.
- DNA extraction procedure was the same as described for PCR (Dellaporta et al, 1983, Plant Mol Biol Rep 1 :19-21 , incorporated herein by reference) with an additional two-time phenol (pH 8.0) extraction and RNase treatment for 30 minutes. Genomic DNA was restricted with Hindlll and Sstl (PR10.1) or Hindlll and EcoRI (chitinase, defensin and DRR206).
- the UV crosslinked Zeta-probe blotting membrane was hybridized with 32 P-labeled random primed probe in 0.5 M Na 2 HP0 4 , pH 7.2 and 7% SDS at 65°C. A 0.5 kb fragment containing the Nptll gene from pBI121 was used as a probe.
- DNA extraction was performed according to a modified protocol from Dellaporta, described above. Leaves (0.5 g) were added to 500 ⁇ l of extraction buffer (100 mM Tris-HCI (pH 8.0), 50 mM EDTA, 500 mM NaCl, 1.25% SDS) and incubated for 20 minutes at 65°C. Next, KOAc was added to a final concentration of 1 M. The solution was kept on ice for 20 minutes and then extracted with chloroform:isoamyl alcohol (24:1 ). For the PCR reactions, mixture contained 1.5 mM MgCI, 0.2 mM dNTP, 0.5 ⁇ M of the specific primer mix and 2.5 units of Taq DNA polymerase.
- extraction buffer 100 mM Tris-HCI (pH 8.0), 50 mM EDTA, 500 mM NaCl, 1.25% SDS
- KOAc was added to a final concentration of 1 M.
- the solution was kept on ice for 20 minutes and then extracted with chloroform
- Gene-specific primers were as follows: for DRR206: OC206+1 (aattccaaacaagagaaagcc) and oC206-2 (cttgatataaacaccaagtcg); for defensin: oS39b+3 (caagaaatagtggtgagtgaa) and oS39b-4 (gcgacaaccacgtgattttg).
- PCR cycles as follows: first cycle: 94°C for 3 minutes, 55°C for 45 seconds, 72°C for 1 minute; 2-29 cycles: 94°C for 45 seconds, 55°C for 45 seconds and 72°C for 1 minute.
- extraction buffer 4 M Guanidine isothiocyanate, 0.1 M Tris-HCI, pH 7.5, 10 mM EDTA, 1 % mercaptoethanol
- Drr230 Choang and Hadwiger, 1991 ) were cloned into T-DNA-based binary vector pB1121 , replacing the GUS gene, as shown in Figure 1 and as discussed above.
- Constructs were directly transfected into A. tumefaciens, and cotyledonary petioles were transformed by cocultivation, as described above.
- T1 plants was pooled and expression of transgenes assayed by RNA gel blot analysis, as described above.
- cotyledons of 20 8-day seedlings were grown for each line, and inoculated with compatible blackleg PG2.
- Transgenic lines containing PR10.1 exhibited similar disease scores to the Westar control, and mRNA levels for this gene ranged from high to undetectable.
- the transformed lines without mRNA accumulation were used as controls for disease scoring, in addition to the untransformed, as shown in Table 1 and Figure 3.
- Among three lines containing chitinase constructs only GN2-2#13 and GN2-2#31 expressed chitinase mRNA. These lines showed only a small decrease in adult plant disease. The strongest effect was seen with DRR206. Five lines exhibiting DRR206 expression also showed significantly lower disease scores, compared to lines with no detectable expression.
- T2 plants from GN3-4#22 also showed lower disease scores in the cotyledon assay. Of two lines carrying pea defensin, the one line that exhibited mRNA expression for this gene also had slightly lower disease scores in adult and cotyledon assays with both T1 and T2 plants.
- T1 lines with lowest disease scores GN3-4#22 (DRR206) and GN4-2#15 (defensin) were self- propagated.
- T2 plants containing the genes were identified by PCR analysis using primers specific for pea DRR206 and defensin.
- T2 plants from the GN3-4#22 line with the DRR206 gene uniformly exhibited significantly improved resistance to L. maculans when compared to T1 plants and to the Westar control. Their cotyledon disease scores were only 2.6 ⁇
- 0.5 and adult disease scores were 1.4 ⁇ 0.6, as shown in Table 2.
- T2 plants from the GN4-2#15 line with defensin did not show significantly more resistance than T1 progenitors.
- the disease scores of cotyledon and adult plants were 7.2 ⁇ 0.5 and 3.4 ⁇ 0.9 respectively, as shown in Table 2.
- Infection phenotypes of Westar with defensin and DRR206 transgenic plants were also compared.
- a large area of the cotyledon was infiltrated with PG2 pycnidiospores using a syringe with no needle.
- untransformed Westar plants exhibit browning, indicative of necrosis, at 8 days post-inoculation.
- Defensin transgenics also exhibit necrosis.
- DRR206 transgenics exhibit chlorosis or yellowing of inoculated tissue, indicative of a hypersensitive response.
- DRR206 protein expression The expression of the DRR206 protein was verified by protein gel blots, as described above and as shown in Figure 5, using polyclonal antibodies from rabbits injected with DRR206 expressed in E. coli
- pea expresses a basic resistance to F. solani f. sp. phaseoli, along with a strong induction of DRR206 mRNA (Fristensky et al, 1985).
- DRR206 polypeptide inferred from the complete sequence of the genomic clone, is 20.4 kD, while the molecular weight of the purified peptide was previously shown to be 23 kD (Culley et al, 1995b).
- the 28 kD band was also faintly visible in proteins from untransformed Westar and is therefore presumably an unrelated protein that also cross-reacts with the antisera. Neither the 25 kD nor 22 kD proteins were visible in Westar-derived protein extracts. Proteins from T1 and T2 resistant DRR206 transgenic plants exhibited bands which co-migrated with the 25 kD and 22 kD bands from pea. The presence of two bands in pea and transgenic
- Brassica suggests that DRR206 undergoes post-translational modifications.
- Botany, Univ. of Toronto, Toronto, Canada were incubated on potato dextrose agar (PDA) for 3 days at room temperature.
- Mycelial plugs (3 mm) were cut and transferred to new PDA plates and incubated at 24°C, 48 hr.
- Fresh plugs containing new mycelia from the growing margin of each colony were cut and placed in the middle of the first two leaves of each plant.
- Plants were incubated in a mist chamber for 2 days. The misting chamber was located in a growth room and illuminated with ambient light. The diameters of necrotic lesions were measured after 48 hr.
- Figure 7 shows the diameter of lesions on ⁇ . napus plants inoculated with Sclerotinia. On untransformed cultivar Westar, the mean lesion size is 2.2 cm
- 4#22-2 a mean lesion size of 1.5 cm (mean of 20 plants inoculated on 4 leaves).
- DRR206 transgenic plants were inoculated after 8 day planting and disease scores were tallied at 8 days postinoculation. "Plant death” was typically obvious, with lodging and complete wilting of all above-ground parts of the seedling, as shown in Table 6. As can be seen, transgenic lines were much more resistant to Rhizoctonia solani compared to the Westar control. The significance of these observations is that DRR206-mediated resistance is clearly not specific for a single fungal isolate, but is effective against a range of fungal species with different modes of pathogenicity, as discussed below.
- the gene influences the expression of resistance in the host, either through direct (epistatic) effects on defense regulatory pathways, or as an indirect (pleiotropic) effect leading to the induction of defense pathways.
- the gene product provides some function that is compatible with the host's defenses, but is normally lacking in the incompatible interaction.
- the pea chitinase (Pissa;Chia;1 ;2) used in this paper is a PR3 endochitinase in the same subfamily as the bean chitinase (Phavu;Chia;1 ;3) used previously (Broglie et al, 1991). Specifically, these proteins share 87% amino acid identity.
- Constitutive expression of the bean chitinase decreased seedling mortality in both transgenic tobacco and B. napus cv. Westar to Rhizoctonia solani (Broglie et al, 1991 ). In our work, the same cultivar of B. napus was used. The simplest explanation therefore is that L. maculans is less sensitive to suppression by PR3- type chitinases than R. solani.
- DRR206 is induced strongly by an incompatible race of F. solani, and more weakly by a compatible race (Fristensky et al, 1985). With compatible isolates of P. syringae, an early induction of DRR206 is seen, which subsides after 12 h.p.i. In gene-specific incompatible interactions with P. syringae, DRR206 is induced early and expression remains strong for at least 30 h.p.i (Daniels et al, 1987). In F. oxysport/m-infested soil, DRR206 is very strongly expressed in resistant isoline
- DRR206 transgenic plants appear to express a hypersensitive response (HR) to fungal inoculation as evidenced by chlorosis and tissue collapse (Figure 4k), despite the fact that the 35S-DRR206 gene itself is not inducible.
- HR hypersensitive response
- Figure 4k chlorosis and tissue collapse
- Pea defensin is less effective than DRR206 in conferring resistance to L. maculans. Extracts from defensin-transgenic plants inhibit spore germination and growth. This is consistent with previous reports showing that radish defensin can cause hyper-branching and decreased growth of Alternaria longipes in-vitro
- DRR206-mediated resistance is not specific for a single fungal isolate but is effective against a range of different fungi with different modes of pathogenicity.
- blackleg fungus is a hemibiotroph whereas Scleotinia is a necrotroph.
- blackleg infects above- ground portions of the plant whereas Rhizoctonia is a root pathogen.
- DRR206 gene comes from pea, and is activated by both bacteria and fungi in pea. Consequently, there is no reason to believe that the effectiveness of DRR206 would be limited to Brassica napus. Clearly, support is shown for the effectiveness of DRR206- mediated resistance in many plant species, against many pathogens.
- inducible promoters may be used in the construction of vectors for expression of plant defense genes, for example, DRR206 and defensin.
- the inducible promoters may be responsive to pathogen attack.
- pathogen-inducible promoters include the pea DRR49a promoter (Culley, D. et al. (1995) Plant Physiology 109:722), the pea DRR206c promoter (Wang, Y. et al., (1999) Molecular Plant-Microbe Interactions 12:410-418), the tobacco PR-1a promoter (Strompen, G.
- Table 1 Specific RNA accumulation and disease scores in canola transformed lines with defense genes. All plants are T1 transformants, except where indicated as T2.
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Abstract
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AU44945/99A AU4494599A (en) | 1998-07-03 | 1999-07-02 | Method for genetic engineering of disease resistance using the drr206 class of proteins |
CA002337095A CA2337095A1 (fr) | 1998-07-03 | 1999-07-02 | Recombinaison genetique de la resistance aux maladies au moyen de la classe de proteines drr206 |
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CA 2242116 CA2242116A1 (fr) | 1998-07-03 | 1998-07-03 | Methode de genie genetique liee a la resistance a la maladie utilisant la categorie de proteines dr206 |
US9175198P | 1998-07-06 | 1998-07-06 | |
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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EP1534829A2 (fr) * | 2002-06-21 | 2005-06-01 | E.I. Du Pont De Nemours And Company | Defensines vegetales |
WO2009043182A1 (fr) * | 2007-10-05 | 2009-04-09 | University Of Manitoba | Marqueurs de région amplifiée de séquence connue (scar) pour la résistance de plantes à des pathogènes fongiques |
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WO1993005153A1 (fr) * | 1991-08-29 | 1993-03-18 | Zeneca Limited | Proteines biocides |
WO1993019188A1 (fr) * | 1992-03-20 | 1993-09-30 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Gene chimerique sensible aux champignons |
US5312912A (en) * | 1989-06-13 | 1994-05-17 | Hadwiger Lee A | Procedures and regulatory DNA sequences for genetically engineering disease resistance and other inducible traits in plants |
WO1997041237A1 (fr) * | 1996-04-29 | 1997-11-06 | COMMONWEALTH SCIENTIFIC AND INDUSTRIAL RESEARCH ORGANISATION (as a participant in the COOPERATIVE RESEARCH CENTRE FOR TROPICAL PLANT PATHOLOGY) | Plantes transgeniques resistantes aux champignons |
WO1998026083A1 (fr) * | 1996-12-13 | 1998-06-18 | Monsanto Company | Polypeptide antifongique et procedes de lutte contre les champignons pathogenes des plantes |
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1999
- 1999-07-02 AU AU44945/99A patent/AU4494599A/en not_active Abandoned
- 1999-07-02 WO PCT/CA1999/000608 patent/WO2000001824A2/fr active Application Filing
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US5312912A (en) * | 1989-06-13 | 1994-05-17 | Hadwiger Lee A | Procedures and regulatory DNA sequences for genetically engineering disease resistance and other inducible traits in plants |
WO1993005153A1 (fr) * | 1991-08-29 | 1993-03-18 | Zeneca Limited | Proteines biocides |
WO1993019188A1 (fr) * | 1992-03-20 | 1993-09-30 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Gene chimerique sensible aux champignons |
WO1997041237A1 (fr) * | 1996-04-29 | 1997-11-06 | COMMONWEALTH SCIENTIFIC AND INDUSTRIAL RESEARCH ORGANISATION (as a participant in the COOPERATIVE RESEARCH CENTRE FOR TROPICAL PLANT PATHOLOGY) | Plantes transgeniques resistantes aux champignons |
WO1998026083A1 (fr) * | 1996-12-13 | 1998-06-18 | Monsanto Company | Polypeptide antifongique et procedes de lutte contre les champignons pathogenes des plantes |
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CULLEY, DAVID E. ET AL: "Molecular characterization of disease-resistance response gene DRR206 -d from Pisum sativum (L.)" PLANT PHYSIOL. (1995), 107(1), 301-2 , XP002125565 -& CULLEY, D.E., ET AL.: "Pisum sativum disease resistance response protein 206-d (DRR206-d) gene, complete cds" EMBL ACCESSION NO: U11716, 15 July 1994 (1994-07-15), XP002125566 * |
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US AN 1999:33133, WANG, Y., ET AL.: "Expression of native and constitutively-expressed pea defense genes in transgenic plants" XP002125573 & CANADIAN JOURNAL OF PLANT PATHOLOGY, (MARCH, 1988) VOL. 20, NO.1, PP131. MEETING INFO: ANNUAL MEETING OF THE CANADIAN PHYTOPATHOLOGICAL SOCIETY WINNIPEG, MANITOBA, CANADA JULY 1997, * |
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1534829A2 (fr) * | 2002-06-21 | 2005-06-01 | E.I. Du Pont De Nemours And Company | Defensines vegetales |
EP1534829A4 (fr) * | 2002-06-21 | 2007-08-22 | Du Pont | Defensines vegetales |
WO2009043182A1 (fr) * | 2007-10-05 | 2009-04-09 | University Of Manitoba | Marqueurs de région amplifiée de séquence connue (scar) pour la résistance de plantes à des pathogènes fongiques |
Also Published As
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WO2000001824A3 (fr) | 2000-02-24 |
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