紫杉木霉菌株ZJUF0986原生质体的制备及诱变抗性菌株的选育  

Protoplast preparation of Trichoderma taxi ZJUF0986 and screening of resistant mutants

作　　者：王国平[1] 周转忠 陈雄燕[2] 郑必强[1] 章初龙[2] 

机构地区：[1]浙江省建德市大洋化工有限公司,建德311616 [2]浙江大学生物技术研究所,杭州310029

出　　处：《植物保护学报》2009年第5期397-402,共6页Journal of Plant Protection

基　　金：浙江省重大科技攻关项目(2006C12008)

摘　　要：通过原生质体诱变,以目标代谢产物木霉菌素为抗性筛选标记选育木霉菌素高产菌株。紫杉木霉野生型菌株ZJUF0986在N15培养基中生长20h的菌丝体,用15mg/mL崩溃酶、30mg/mL蜗牛酶和30mg/mL溶壁酶组合成的混合酶为酶裂解液,以菌丝量∶酶裂解液=1∶2.5(w/v)的比例,在30℃、100r/min条件下酶解90min,原生质体释放量最佳,达到9.42×108个/mL。所制备的原生质体进行氯化锂-紫外复合诱变,紫外照射剂量为60s时的突变率和正突变率最高,筛选到高产突变体菌株UL60-11,其发酵效价比野生型菌株提高67.3%。Screening high yielding mutants of trichodermin production by protplast mutagenesis with resistance of target metabolite trichodermin was developed. Trichoderma taxi wild strain ZJUF0986 was grown in N15 medium for 20 h and mycelium was harvested. Mycelium was treated with mixed enzyme combined with 15 mg/mL driselase, 30 mg/mL helicase and 30 mg/mL lywallzyme at ratio of mycelium ： enzyme = 1 ： 2.5 （ w/v）, the optimal condition of protoplast release was at 30 ℃, 100 r/min for 90 min and protoplast production reached 9.42 ×10^8/mL. Protoplast of ZJUF0986 was mutated with lithium chloride solution and UV. The mutation rate and positive mutation rate were the highest on 60 s UV radiation dose. A high yield mutant strain UL60-11 was screened, which increased 67.3% of fermentation efficiency than wild strain ZJUF0986.

关 键 词：内生真菌 代谢产物 抗真菌活性 原生质体 氯化锂-紫外诱变 

分 类 号：S476[农业科学—农业昆虫与害虫防治]