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期 刊 ：植物保护学报 2009年

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；

Keywords:endophytic fungi, metabolite, antifungal activity, protoplast, lithium chloride-UV mutation,

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20h的菌丝体,用15mg/mL崩溃酶、30mg/mL蜗牛酶和30mg/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℃、100r/min条件下酶解90min,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60s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%。

Abstract: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 9.42×10⁸/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.

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