1997 Fiscal Year Final Research Report Summary
Creation of Pathogen-resistant Transgenic Plants by Introducing Chitinase Gene
| Project/Area Number |
08660058
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
植物保護
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| Research Institution | Yamaguchi Univeristy |
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Principal Investigator |
KOGA Daizo Yamaguchi University Faculty of Agriculture, Department of Biological Science Professor, 農学部, 教授 (30091185)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Plant self-defense / Chitinase / Transgenic plants |
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| Research Abstract |
Plants induce a series of pathogenesis-related proteins (PR-proteins) in response to the attack by plant pathogens. Chitinase is a major PR protein. In this study, we have investigated the application of chitinase for creation of pathogen-resistant transgenic plants by introducing chitinase gene.
(1) Chitinase is classified into two families of glycosyl hydrolase such as family 18 and 19. Plant chitinse is also classified into five classes. We purified and characterized several chitinase isozymes from yam tuber and the silkworm, Bombyx mori. As the result, we could clarify that chitinase isozymes with strong lytic activity against pathogens belong to class III and IV and also family 18. Furthermore, we found that such isozymes are due to their affinity to solid chitin.
(2) We analyzed nucleotide sequence and amino acid sequence of the class IV chitinase from yam. Furthermore, we constructed the chimeric DNA by adding the signal sequece of sugar beet class IV chitinase to the DNA encoding yam class IV chitinase. We introduced this constructed gene into tobacco and strawberry by using the binary system of Agarobacteriium tumefaciens. We succeeded in obtaining the transgenic plants carrying yam chitinase gene and reveaing the pathogen-resistance.
(3) We found the important points that E.coli and Agrobacterium tumefaciens which were used for introduing chitinase gene and carried this gene could not easily grow. We are now resolving this problem by using particle gun instead of Agrobacterium system, and by inventing the plan in the construction of chitinse gene.
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