摘要
研究可降解多菌灵的木霉菌株Tr1的原生质体制备及再生的适宜条件,发现该菌原生质体制备的最佳条件为菌龄20 h,裂解酶液为5 mg/m L的溶菌酶和蜗牛酶的混合酶液,二者使用前按照体积比2∶1混合,酶解温度30℃,50 r/min缓慢振荡酶解2.5 h,渗透压稳定剂为0.7 mol/L KCl,原生质体产量达6.44×106个/m L。原生质体再生的优化条件为:PDA为培养基,添加0.3 mol/L KCl和0.3 mol/L环己六醇作为渗透压稳定剂,采用双层固体培养方式,有助于原生质体的再生。
The suitable preparation and regeneration conditions of protoplast preparation of Trichoderma sp. Tr1 was studied.The results showed the optimum condition for preparation of protoplast of strain Tr1 ：The culture time of mycelium was 20 h,the protoplast lysate was a mixed solution of 5 mg / m L lysozyme and 5 mg / m L snail enzyme and they would be mixed according to the volume ratio of 2∶1 before using, it was mildly shaked with a speed of 50 r / min under 30 ℃ for 2.5 h, 0.7 mol / L KCl was used as osmotic stabilizer. Under this condition, the yield of protoplast could be 6.44×10^6/ m L. The optimum condition for regeneration of protoplast was PDA as the basic culture medium,0.3 mol / L KCl and 0.3 mol / L inositol as the osmotic stabilizer, and double-layer solid culture was helpful for protoplast regeneration of Trichoderma sp. Tr1.
出处
《湖北农业科学》
2016年第10期2500-2503,共4页
Hubei Agricultural Sciences
基金
山东省科技发展计划项目(2013GNC11019
2012GGC01020
2014GSF121028)
国际科技合作项目(2010DFA32330)
关键词
多菌灵
木霉
原生质体
制备
再生
carbendazim
Trichderma sp.
protoplast
preparaton
regeneration
