根结线虫生防真菌交枝顶孢原生质体的制备与再生体系构建

Preparation and regeneration of protoplasts from the biocontrol fungus Acremonium implicatum

本研究以根结线虫生防真菌交枝顶孢Acremonium implicatum为供试菌株,探究了菌龄、降解酶种类、酶解温度和时间、不同渗透压稳定剂、不同pH等因素对其原生质体制备的影响,并建立了交枝顶孢原生质体制备体系;制备的原生质体在SR培养基上再生率达到17.78%。此外,GFP遗传转化菌株的获得充分表明制备的原生质体可作为后续遗传转化材料,为进一步研究交枝顶孢对根结线虫的生防机理奠定基础。

In order to establish protoplast-mediated genetic transformation of the biocontrol fungus Acremonium implicatum,the conditions for the isolation and regeneration of A.implicatum protoplasts were optimized,including culture time,enzyme system,enzymatic digestion time and temperature,type of osmotic stabilizers,pH value and so on.For the preparation of protoplasts,solid regeneration medium was better with a regeneration rate of protoplasts of 17.78%.The transformant strain with GFP was obtained.The system may build a foundation for further study on the mechanism of biocontrol against the root-knot nematode.