果生刺盘孢侵染苹果致病关键基因Cfcyp450的功能  

Function of cytochrome P450 gene critical for Colletotrichum fructicola infection to apple

作　　者：陈金珠 王静钶 刘广利 卢一鸣 梁晓飞[1] 孙广宇[1] 张荣[1] CHEN Jinzhu;WANG Jingke;LIU Guangli;LU Yiming;LIANG Xiaofei;SUN Guangyu;ZHANG Rong(College of Plant Protection,Northwest A&F University,Yangling 712100,Shaanxi,China)

机构地区：[1]西北农林科技大学植物保护学院,陕西杨凌712100

出　　处：《菌物学报》2023年第3期770-781,共12页Mycosystema

基　　金：国家自然科学基金(32070144,32072374);国家重点研发计划(2019YFD1002002)。

摘　　要：果生刺盘孢Colletotrichum fructicola能够引起苹果炭疽叶枯病和苹果苦腐病。前期通过基因组分析发现一个叶枯型C.fructicola菌株特异基因Cfcyp450。本研究对其功能进行分析,以明确其在致病中的作用。采用同源重组方法获得了敲除突变体。表型分析显示,Cfcyp450敲除突变体与野生型生长速率无明显差别,但敲除突变体菌落变白;与野生型相比,突变体附着胞形成降低30.02%;侵染钉延伸菌丝对玻璃纸的穿透率下降41.19%。致病性测定显示,突变体对嘎啦叶片致病力显著下降。生物信息学分析显示,Cfcyp450仅存在于叶枯型菌株中,同时与烟草和拟南芥内生菌同源性较高,但与刺盘孢属物种亲缘关系较远。这些结果表明果生刺盘孢Cfcyp450基因为叶枯型菌株的关键致病因子,可能通过水平转移获得。Colletotrichum fructicola is the dominant pathogen of apple Glomerella leaf spot(GLS)in China.In this study,the function of putative cytochrome P450 gene,Cfcyp450,specific in C.fructicola GLS pathotype was analyzed.Phenotypic analysis showed that deleting Cfcyp450 did not affect vegetative growth rate of C.fructicola,but the vegetative colony of the knockout mutant turned white.Compared to the wild type,appressorium formation rate reduced by 30.02%,and cellophane penetration rate reduced by 41.19%.The virulence to apple leaves was obviously decreased.Sequence analysis showed that Cfcyp450 was only found in GLS pathotype of C.fructicola and has a higher homology with cyp450 in endophytes of tobacco and Arabidopsis thaliana,but has a distant genetic relationship with cyp450 in other Colletotrichum spp.Our results reveal that the gene Cfcyp450 is the key pathogenic gene of C.fructicola,and could be obtained by horizontal gene transfer.

关 键 词：细胞色素P450 豌豆素脱甲基酶 效应因子 叶枯型 基因水平转移 苦腐病 

分 类 号：S436.611.12[农业科学—农业昆虫与害虫防治]