| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1990: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1989: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1988: ¥700,000 (Direct Cost: ¥700,000)
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| Research Abstract |
Conditions for isolation, culture, and fusion of protoplasts and separation of DNA into chromosome size molecular in entomopathogenic fungi were studied. High yields of protoplasts from young mycelia of some entomopathogenic fungi were obtained by treatment with Zymolyase or Novozyme 234. In protoplast fusion between the auxotrophic mutants of Paecilomyces fumosoroseus the complementation frequencies were 10^<-1>-10^<-2>
To use the protoplasts of entomogenous fungi more easily and effectively for genetic purposes, proper techniques for protoplast preservation are needed. However, little attempt has been made to preserve protoplasts of entomopathogenic fungi. We then tested glycerol as a cryoprotectant at concentrations 0, 20, 40, 60, and 80%. After 2days of storage at -80^ﾟC, the best protection was obtained at the concentration of 20%, as judged by the viability of the protoplasts. When the value of the untreated control was taken as 100%, those of 21 and 48 days of storage were as 71.3% and 68.8%, respectively.
The genetics of entomopathogenic fungi is in its fancy and it is not even certain how many chromosomes these fungi contain. We tested the separation of P. fumosoroseus protoplast DNA into chromosome size molecules by pulsed field electrophoresis. The P. fumosoroseus genome was solved into six bands. The bands were estimated, in descending order, to be 7.8, 6.2, 5.3, 4.4, 3.3 and 3.1mb. The Total P. fumosoroseus gennome is therefoe more than 30.1mb.
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