捕食性真菌Duddingtonia flagrans原生质体的快速制备与再生

Rapid preparation and regeneration of protoplast from nematode-trapping fungus Duddingtonia flagrans

为快速获得大量捕食性真菌Duddingtonia flagrans原生质体并使其再生,本试验研究了酶质量浓度、酶解温度、菌龄、酶解时间对D.flagrans原生质体产生的影响,以确定D.flagrans原生质体最佳快速制备条件。结果表明,在溶壁酶2mL/L、蜗牛酶8g/L、纤维素酶8g/L时,35℃恒温条件下,酶解菌龄为2d的菌丝7h,捕食性真菌D.flagrans产生原生质体数量最多且能够再生。这为下一步转化并标记和克隆捕食性相关基因奠定了重要基础。

In order to obtain a large number of protoplasts of nematode-trapping fungus-Duddingtonia flagrans,and optimize the factors that affect protoplast generation,the effects of the enzyme concentration,reaction temperature of the enzyme,age of culture and incubation time on thepreparation of protoplast of D.flagrans were observed.The results showed that,a large number of protoplasts were generated when the mycelia of D.flagrans grown for 2days were treated with 2mL/L lywallzyme,8g/L glusulase and 8g/L cellulose in 35℃for 7h,and some viable protoplasts could regenerate dmycelia.The trial primarily ascertained the optimum conditions to prepare the protoplast of the fungus,which supplied a foundation for transforming,marking and cloning the genes of the nematode-trapping fungi.