2007•Xi'nan nongye xuebaoRequires access
Production and regeneration of Trichoderma protoplast.
Yutao Cao, Peng HuaXian, Wen Cheng-jing, Bowei Liu, Xi YaDong
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Abstract
This paper reported that mycelia age,enzyme proportion,osmotic stabilizer-buffer systems,enzymolysis time as well as medium composition effect on protoplast preparation and regeneration of Trichoderma strain T25 and T55.The results showed that optimal conditions of protoplast preparation of T25 and T55 were osmotic stabilizer-buffer systems composed of 13 mg/mL driselase,0.2 mol/L PBS,pH5.8 and 0.6 mol/L NaCl.T25 Protoplast was obtained from 20 h-old mycelia after 3 h treatment at 30 ℃ and the number of the protoplast was about 9.0×106 cfu/mL.T55 protoplast was obtained from 26h-sold mycelia after 3-hour treatment at 30 ℃ and the number of the protoplast was about 6.33×106 cfu/mL.The medium of protoplast regeneration of T25 and T55 composed of Czapek medium containing 0.5 % yeast extract and 0.5 % Calium pantothenic with NaCl.The regeneration rate was 1.150 % and 0.785 %.