玉米大斑病菌原生质体遗传转化体系的建立

Establishment of the Genetic Transformation System of Protoplasts from Setosphaeria turcica

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 Setosphaeria turcica strain 01-23 was used to study main conditions which had impacl on protoplast genetic transformation, including the materials of protoplast preparation, the enzyme system, the osmotic stabilizer, the concentration and linearized degree of vector and so on. The results indicated that the conidial suspension incubated for 24 h was the optimal material for the preparation of protoplasts. The mixture of 2% Lywallzyme and 1% Snailase was the best enzyme system. 0.7 mol/L NaC1 was the most appropriate osmotic stahilizer used for the transfor mation of protoplast from S. turciea. The transformation efficiency of cyclic vector was five times higher than the linear vector. The transformation efficiency reached the highest when the carrier concentration was 100 μg/mL.