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原生质体质量是丝状真菌遗传转化成功的关键。本研究以甘蔗黑穗病菌为试验材料,对其原生质制备和再生的条件进行研究。结果表明,将甘蔗黑穗病菌单倍体菌株JG36摇床培养24 h,然后加入5 mg/m L溶壁酶、30 mg/m L纤维素酶和30 mg/m L蜗牛酶的混合酶液,28℃160 r/min摇床上酶解3 h。1.2 M/L Mg SO_4作为原生质体制备时的渗透压稳定剂。在此条件下,制备原生质体得率最高达86.86%,原生质体再生率达到63.26%。
Protoplast quality is the key to successful transformation of filamentous fungi. In this study, sugarcane smut was used as experimental material to study the preparation and regeneration conditions of its protoplasts. The results showed that the haploid strain JG36 of Saccharomyces cerevisiae was cultured for 24 h and then mixed enzyme solution of 5 mg / ml L-lysozyme, 30 mg / ml cellulase and 30 mg / ml snail enzyme , 28 ℃ 160 r / min shaker 3 h. 1.2 M / L Mg SO_4 as protoplast preparation osmotic pressure stabilizer. Under these conditions, the yield of protoplast was up to 86.86%, protoplast regeneration rate reached 63.26%.