Title Trichoderma koningii의 種間 原形質體 融合에 대한 硏究
Author 洪淳佑 · 河永七 · 朴喜門 · 趙南鎭
Address 서울大學校 自然科學大學 微生物學科
Bibliography Korean Journal of Microbiology, 22(2),103-110, 1984
Key Words Intraspecific Protoplast Fusion in Trichoderma koningii
Abstract The conditions for the protoplast fusion of auxotrophic mutants of Trichoderma koningii were determined. A preparation of commercial enzyme Driselase was used successfully to isolate protoplasts from the 18 hr old mycelium of T. koningii. The yields of protoplasts production were ranged from 0.3x10^8 to 2.5x10^8 protoplasts per mg of damp mycelium of various auxotrophic mutant strains. The regeneration frequencies from 9.3x10^-3 to 2.0x10^-1 were obtained when the protoplasts from auxotrophic mutants were plated on the malt extract medium containing 0.6M MgSO_4, and 2% agar, and the optimal concentration of PEG for protoplst fusion was 30%. Exposure of protoplasts to PEG for 10 min was found to be sufficient to induce high frequency heterokaryon formation. Optimal pH of fusion mixture was determined as 5.5, and 1 mM of calcium chloride in fusion mixture was found to be sufficient to enhance protoplast fusion frequency. Under optimal condition, the fusion frequency of the cross between protoplasts from various auxotrophic mutants were 1.6x10^-2 and 4.1x10^-2.
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