莲腐败病菌遗传转化体系的构建

由共享镰刀菌Fusarium commune引起的莲腐败病是我国莲生产上的主要病害之一。本研究以强致病力菌株FCN23为供试菌株,通过PEG介导的遗传转化法研究了菌龄、酶解时间和渗透压稳定剂等对原生质体制备的影响。结果表明,分生孢子在YPD液体培养基培养24 h获得新鲜菌丝,在酶解时间为2.5 h,渗透压稳定剂为0.7 mol/L NaCl溶液时原生质体制备效率最高。进而将遗传霉素的抗性基因和绿色荧光蛋白基因转入莲腐败病菌原生质体中，获得了稳定表达的转化子,能够稳定遗传gfp基因,说明莲腐败病菌的遗传转化体系构建成功。该体系的建立为莲腐败病菌的侵染过程及基因功能研究奠定了基础。

Construction of genetic transformation system of Fusarium commune causing lotus rhizome rot

Lotus rhizome rot caused by Fusarium commune is one of the major diseases in lotus production in China. In this study, the strongly pathogenic strain FCN23 was used as the target strain, and the effects of enzymatic digestion time, mycelial age and osmotic stabilizer on the protoplast preparation were investigated by PEG-mediated genetic transformation. When the conidia were cultured in YPD liquid medium for 24 h to obtain fresh mycelium, the enzyme digestion time was 2.5 h, the osmotic stabilizer was 0.7 mol/L NaCl solution, and the efficiency was the highest. Geneticin resistance gene and green fluorescent protein gene were transfered into the protoplast of F.commune. The results showed that stably expressed transformants were obtained, which were able to stably inherit the gfp gene, indicating that the genetic transformation system of F.commune was successfully constructed. The transformation system laid the foundation for the study of the infestation process and gene function of F.commune.