论文部分内容阅读
以玉米大斑病菌菌株01-23为出发菌株,对影响原生质体遗传转化的主要条件进行了研究,包括原生质体制备的材料、酶解系统、稳渗剂、载体浓度及线性化程度等。结果表明,改良Fries培养基中培养24 h的分生孢子悬浮液是原生质体制备的首选材料;2%溶壁酶和1%蜗牛酶两种酶混用是最佳的酶解系统;0.7 mol/LNaCl是最适的稳渗剂;环状载体比线性载体转化效率高5倍以上;在载体浓度为100μg/mL时转化效率最高。
The main influencing factors on the genetic transformation of the protoplast were studied, including protoplast preparation, enzymolysis system, stabilizing agent, concentration of carrier and linearity. The results showed that the conidiospore suspension cultured in modified Fries medium for 24 h was the preferred material for the preparation of protoplasts. The optimum enzymatic hydrolysis system was the mixture of 2% lyse and 1% LNaCl is the most suitable stabilizing agent; the cyclic vector is more than 5 times more efficient than the linear vector; and the highest transformation efficiency is obtained when the carrier concentration is 100μg / mL.