【摘要】 通过不同菌龄、酶液、缓冲液?稳定剂系统、酶解时间、再生培养基组合等条件对木霉T25、T55原生质体制备和再生的影响研究,结果表明,木霉T25、T55原生质体制备和再生的最适条件为:崩溃酶13 mg/mL,0.2 mol/L磷酸缓冲液(PBS),pH5.8和0.6 mol/L NaC l组成的缓冲液稳定系统,木霉T25菌龄为20 h,木霉T55菌龄为26 h的菌丝体在30℃下,酶解3h后,可分别获得原生质体数量9.0×106个/mL、6.33×106个/mL,两菌的再生培养基都是以加入0.5%酵母膏和0.5%泛酸钙的改良Czapek与NaC l的组合培养基,T25与T55的再生率分别为1.150%和0.785%。更多还原

【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 %.更多还原