中文摘要：

为验证苹果树腐烂病菌VmKu80的基因功能,构建高效率基因敲除的受体菌株。通过基因敲除技术获得介导非同源末端连接（NHEJ）途径的VmKu80基因缺失突变体,并对其生长速率、皿内菌落形态、致病力、分生孢子器的形成和基因敲除效率等进行分析。结果表明,该突变体的生长速率、皿内菌落形态、对苹果的致病力和分生孢子器产生等均与03-8菌株无显著差异,且以VmKu80缺失突变体为受体菌株的基因敲除效率是03-8菌株的12倍。说明,VmKu80缺失突变体的获得可以实现苹果树腐烂病菌基因敲除工作的高通量化,有助于苹果树腐烂病菌的基因功能研究。

英文摘要：

This study is to verify the gene function of VmKu80 which mediates the NHEJ pathway and construct the recipient strain with improved frequency of targeted gene replacement of V.mali.VmKu80 gene knockout mutant named ΔVmKu80 was obtained by CaCl_2 ·PEG mediated genetic transformation.The growth speed,colony morphology,pathogenicity and pycnidium formation were studied,the results showed no significant difference with wild type 03-8.Meanwhile,the frequency of targeted gene replacement ofΔVmKu80was 12 fold to wild type 03-8.ΔVmKu80can realize the high throughput genes knockout and promote gene function of V.mali.