Title:
METHOD FOR GENETIC ENGINEERING OF DISEASE RESISTANCE USING THE DRR206 CLASS OF PROTEINS
Document Type and Number:
WIPO Patent Application WO2000001824
Kind Code:
A3
Abstract:
To identify genes effective against the blackleg fungus Leptosphaeria maculans, we have transformed canola (Brassica napus with four pea (Pisum sativum) genes under constitutive control by the CaMV 35S promoter: PR10.1, chitinase, DRR206 and defensin. Transgenic lines containing single copy T-DNA insertions for each gene were screened for both seedling (cotyledonary) and adult plant resistance. Lines for which pea DRR206 or defensin mRNA was expressed also showed decreased disease scores when inoculated with L. maculans or Sclerotinia sclerotiorum, compared to non-expressing transgenic lines. For PR10 and chitinase transgenics, there was little or no enhancement of resistance. Furthermore, resistance to L. maculans co-segregated with DRR206 and defensin transgenes. Extracts from DRR206 and defensin transgenic plants inhibited fungal growth in-vitro. DRR206 transgenic plants also demonstrated decreased hyphal growth at inoculation sites, and evidence of a hypersensitive response.
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Inventors:
FRISTENSKY BRIAN (CA)
WANG YAPING (CA)
WANG YAPING (CA)
Application Number:
PCT/CA1999/000608
Publication Date:
February 24, 2000
Filing Date:
July 02, 1999
Export Citation:
Assignee:
UNIV MANITOBA (CA)
FRISTENSKY BRIAN (CA)
WANG YAPING (CA)
FRISTENSKY BRIAN (CA)
WANG YAPING (CA)
International Classes:
C07K14/415; C12N9/24; C12N15/29; C12N15/82; (IPC1-7): C12N15/29; A01H5/00; A01N65/00; C12N5/10; C12N15/82
Domestic Patent References:
| WO1998026083A1 | 1998-06-18 | |||
| WO1993005153A1 | 1993-03-18 | |||
| WO1997041237A1 | 1997-11-06 | |||
| WO1993019188A1 | 1993-09-30 |
Foreign References:
| US5312912A | 1994-05-17 |
Other References:
DATABASE BIOSIS BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; WANG, Y., ET AL.: "Expression of native and constitutively-expressed pea defense genes in transgenic plants", XP002125573
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
DATABASE SCISEARCH WANG, Y.P., ET AL.: "Constitutive expression of pea defense gene DRR206 confers resistance to blackleg (Leptosphaeria maculans) disease in transgenic canola (Brassica napus)", XP002125574
BROWN, S.M., ET AL.: "Sequence conservation of a family of disease response genes among pea species and their expression in transgenic canola", J. CELL. BIOCHEM. SUPPL., vol. 18a, 1994, pages 86, XP000857499
DATABASE SCISEARCH "FIELD TOLERANCE TO FUNGAL PATHOGENS OF BRASSICA-NAPUS CONSTITUTIVELY EXPRESSING A CHIMERIC CHITINASE GENE", XP002107279
FRISTENSKY, B., ET AL.: "cDNA sequences for pea disease resistance response genes", PLANT MOLECULAR BIOLOGY, vol. 11, 1988, pages 713 - 715, XP002125569
CHIANG CH C ET AL: "THE FUSARIUM SOLANI-INDUCED EXPRESSION OF A PEA GENE FAMILY ENCODING HIGH CYSTEINE CONTENT PROTEINS", MOLECULAR PLANT-MICROBE INTERACTIONS, vol. 4, no. 4, 1991, pages 324-331, XP002063334, ISSN: 0894-0282
TERRAS F R G ET AL: "SMALL CYSTEINE-RICH ANTIFUNGAL PROTEINS FROM RADISH: THEIR ROLE IN HOST DEFENSE", PLANT CELL,US,AMERICAN SOCIETY OF PLANT PHYSIOLOGISTS, ROCKVILLE, MD, vol. 7, no. 5, May 1995 (1995-05-01), pages 573-588, XP002029836, ISSN: 1040-4651
MOLONEY, M.M., ET AL.: "High efficiency transformation of Brassica napus using Agrobacterium vectors", PLANT CELL REPORTS, vol. 8, 1989, pages 238 - 242, XP002125570
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
DATABASE SCISEARCH WANG, Y.P., ET AL.: "Constitutive expression of pea defense gene DRR206 confers resistance to blackleg (Leptosphaeria maculans) disease in transgenic canola (Brassica napus)", XP002125574
BROWN, S.M., ET AL.: "Sequence conservation of a family of disease response genes among pea species and their expression in transgenic canola", J. CELL. BIOCHEM. SUPPL., vol. 18a, 1994, pages 86, XP000857499
DATABASE SCISEARCH "FIELD TOLERANCE TO FUNGAL PATHOGENS OF BRASSICA-NAPUS CONSTITUTIVELY EXPRESSING A CHIMERIC CHITINASE GENE", XP002107279
FRISTENSKY, B., ET AL.: "cDNA sequences for pea disease resistance response genes", PLANT MOLECULAR BIOLOGY, vol. 11, 1988, pages 713 - 715, XP002125569
CHIANG CH C ET AL: "THE FUSARIUM SOLANI-INDUCED EXPRESSION OF A PEA GENE FAMILY ENCODING HIGH CYSTEINE CONTENT PROTEINS", MOLECULAR PLANT-MICROBE INTERACTIONS, vol. 4, no. 4, 1991, pages 324-331, XP002063334, ISSN: 0894-0282
TERRAS F R G ET AL: "SMALL CYSTEINE-RICH ANTIFUNGAL PROTEINS FROM RADISH: THEIR ROLE IN HOST DEFENSE", PLANT CELL,US,AMERICAN SOCIETY OF PLANT PHYSIOLOGISTS, ROCKVILLE, MD, vol. 7, no. 5, May 1995 (1995-05-01), pages 573-588, XP002029836, ISSN: 1040-4651
MOLONEY, M.M., ET AL.: "High efficiency transformation of Brassica napus using Agrobacterium vectors", PLANT CELL REPORTS, vol. 8, 1989, pages 238 - 242, XP002125570
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