稻瘟病菌原生质体的制备和再生菌株的致病性

Protoplast Preparation and Pathogenicity of Regenerated Strains of Magnarporthe grisea

以稻瘟病 (Magnarporthe grisea)菌株 ZA49、131为供试菌株 ,进行原生质体的制备试验。结果表明 :用9种酶均能消化该菌细胞壁 ,并获得一定数量的原生质体 ;产生原生质体效率最高的是酶 Novozym TM2 34和Driselase,且这两种酶以 10 mg/ ml浓度产生的原生质体数量最多 ,消化 2～ 3 h后即可获得相当数量的原生质体 ,最适作用温度分别为 30℃和 32℃。以几丁质降解酶与 Novozym TM2 34或 Driselase混用时对制备原生质体有很好的增效作用。作者认为 ,Novozym TM2 34和 Driselase的作用浓度以 5 m g/ ml最为经济、实用。所制备的原生质体均能再生菌丝和具有产生与原菌株相同致病性分生孢子的能力。

The strains ZA49,131 of Magnaporthe grisea were used to prepare protoplasts for transformation of the fungus.Enough protoplasts were obtained by digestion of the fungal cell wall with 9 lytic enzymes among which Novozym TM234 and Driselase had the highest efficiency.The most protoplasts were gained with 10 mg/ml Novozym TM234 or 10 mg/ml Driselase for 2.5～3 h at 30℃ or 32℃.Protoplast preperation responded to the mixture of Chitinase and Novozym TM234 or the one of Chitinase and Driselase.It was proved that 5 mg/ml Novozym TM 234 or 5 mg/ml Driselase is more economic and suitable for preparation of the protoplasts.The prepared protoplasts could regenerate mycelium and produce conidium with similar pathogenecity to that of parental strains.