Analysis of pathogenicity of silkworm pathogenic microorganisms by protoplast fusion and gene technology
| Project/Area Number |
59480050
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
蚕糸学
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| Research Institution | Kyushu University |
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Principal Investigator |
AIZAWA Keio Faculty of Agriculture, Kyushu University, 農学部, 教授 (00038142)
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| Co-Investigator(Kenkyū-buntansha) |
OHBA Michio Faculty of Agriculture, Kyushu University, 農学部, 助手 (80038281)
KANDA Kohzo Faculty of Agriculture, Kyushu University, 農学部, 助手 (00117095)
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| Project Period (FY) |
1984 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1986: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1985: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1984: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Keywords | Beauveria bassiana / Beauveria brongniartii / Bacillus thuringiensis / Insecticidal parasporal inclusion / Protoplast / Protoplast fusion / プラスミド |
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| Research Abstract |
The objective of this research is the application of protoplast fusion and gene technology to the analysis of pathogenicities of insect pathogenic microorganisms against the silkworm, Bombyx mori. Results obtained are as follows:
1. Protoplast fusion in entomognous fungi: Blastospores of Beauveria bassiana were treated with Zymolyase 5000 and high yield of protoplasts of good quality was obtained. Protoplasts from two auxotrophic mutants of B. bassiana requring tryptophan and leucine were treated with polyethylene glycol (PEG) solution added with osmotic stabilizer and <CaCl_2> . The frequency of fusion at 30% PEG (MW 7,500) was about 9 x <10^(-4)> . The frequency of protoplast fusion between B. bassiana (strain requiring tryptophan) and Beauveria brongniartii (strain requiring lysine) was <10^(-4)> - <10^(-5)> and isolates with different combination of markers (colony morphology, amino acid requirements, production of pigment, resistance against benomyl, benzalkonium chloride, and nick
… More
el) were obtained.
2. Plasmid transfer in Bacillus thuringiensis: Using crystalliferous type strain of B. thuringiensis subsp. alesti possessing plasmid pQUl and acrystalliferous (and streptomycin resistant) strain obtained from the the type strain, mating experiments were done to investigate the plasmid transfer. The acrystalliferous strain converted into crystalliferous strain with the acquisition of plasmid pQUl at high frequency (more than <10^(-2)> ) and the existence of pQUl was evidenced to be associated with the production of crystals. The transfer of B. thuringiensis subsp. alesti plasmids into Bacillus subtilis strain Y-12S by mating was also observed but the maintenance of plasmids was not stable. Transformation by pUB110 in protoplasts of B. thuringiensis subsp. dendrolimus type strain was demonstrated.
3. Insecticidal toxin produced by Bacillus thuringiensis: Two toxic proteins were purified from crystals of B. thuringiensis strain PG-14. The mosquitocidal activity was attributed to 65-kD protein and hemolytic activity was due to 25-kD protein. The former protein was purified with good yield by affinity chromatography. Less
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Report
(1 results)
Research Products
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