摘要
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 the transformation of filamentous fungi. In this study, preparation and regeneration of protoplast from Sporisorium scitamineum were studied. The results showed that Sporisorium scitamineum JG36 were cultivated at 28℃, and then incubated in the mixed enzyme liquid consisting of 5 mg/m L lyticase, 30 mg/m L cellase, and 30 mg/m L snailase at 28℃ in a flat shaker(160 r/min) for 3 h. 1.2 M/L Mg SO_4 was used as osmotic stabilizer for protoplast preparation. Under these conditions, the highest preparation rate of protoplast reached to 86.86%, and the protoplast regeneration rate reached to 63.26%.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2016年第7期1750-1755,共6页
Genomics and Applied Biology
基金
国家973计划项目(2015CB150600)资助
关键词
甘蔗黑穗病菌
原生质体
制备
再生
Sporisorium scitamineum
Protoplast
Preparation
Regeneration
