摘 要茶树炭疽病主要是由胶孢炭疽菌引起的重要病害，茶树的生理代谢严重被影响，并且逐年减少茶产量与质量下降。因此，在茶树炭疽病防控中需要根据病原菌的特性制定不同的防控措施。Vps29 是 Retromer 蛋白复合体的亚基之一，可能参与调控病原菌的侵染进程。为此，本研究利用同源重组原理对胶孢炭疽菌VPS29 基因进行了敲除，采用 PCR 扩增 VPS29 基因敲除所需的同源重组片段，转化至胶孢炭疽菌 N425 的原生质体中，通过新霉素抗性筛选转化子以及 PCR 验证得到一株 VPS29 缺失突变菌株命名为 Δvps29-7。对菌株的营养生长和产孢分析的结果表明，在固体培养基上突变体菌丝生长速率小于野生型，说明 VPS29 基因控制胶孢炭疽菌的营养生长。下一步还需进行其它方面的表型分析，以全面鉴定胶孢炭疽菌 VPS29 基因的生物学功能。关键词; 茶树; 胶孢炭疽菌; Retromer 复合体; 基因敲除 Abstract Anthracnose of tea plant is mainly caused by Colletotrichum gloeosporioides, which is the main disease of tea plant. The physiological metabolism of tea is seriously affected by the anthracnose, and this disease reduces the output and quality of tea year by year. Therefore, to prevent and reduce tea anthracnose, different methods were needed according to the characteristics of pathogen.Vps29 is one of the subunits of Retromer protein complex.It may involve in the process of C. gloeosporioides infecting tea trees and have important functions. To this end,In this study, we deleted the VPS29 gene according to the homologous recombination principle in C. gloeosporioides. The VPS29 knockout homologous recombinant fragment was amplified by double-joint PCR and transform to protoplast of C. gloeosporioides N425. Screening of the transformants with geneticin resistance and confirming by PCR. A VPS29 null mutant named Δvps29-7 was generated. Analysis of vegetative growth and sporulation of the Δvps29-7 and we found that the growth of this mutant on solid medium was slo...
