基于Cfku70基因失活策略构建果生刺盘孢的高效基因敲除底盘菌株  

Improving target gene replacement efficiency in Colletotrichum fructicola by inactivating Cfku70 gene

作　　者：田欢焕 杨文睿 张兆辉 姚立强 张荣[1] 孙广宇[1] 梁晓飞[1] TIAN Huanhuan;YANG Wenrui;ZHANG Zhaohui;YAO Liqiang;ZHANG Rong;SUN Guangyu;LIANG Xiaofei(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》2022年第8期1185-1198,共14页Mycosystema

基　　金：陕西省自然科学基金青年项目(2020JQ-255);国家自然科学基金面上项目(32072374,32070144)。

摘　　要：果生刺盘孢侵染危害多种植物,是重要的植物病原真菌。在一些丝状真菌中,敲除非同源末端连接修复通路的关键基因ku70或ku80可显著提高同源重组效率,进而提高靶基因置换频率。本研究从果生刺盘孢基因组鉴定到Cfku70和Cfku80两个基因,并明确了基因失活对菌株生物学表型和基因敲除效率的影响。敲除Cfku70或Cfku80不影响菌株的菌落形态、营养生长、产孢、分生孢子萌发、侵染结构发育和致病;Cfku70基因敲除还大幅提升3个测试基因的敲除效率。本研究证实Cfku70基因失活能显著提高果生刺盘孢的基因敲除效率,适宜作为高效基因敲除的底盘菌株,研究结果为通过批量敲除策略筛选新型致病因子奠定重要基础。Colletotrichum fructicola is a devastating fungal phytopathogen that infects and damages many plants.In certain filamentous fungi,deleting ku70 or ku80,key genes of the nonhomologous end joining(NHEJ)pathway,can significantly boost homologous recombination and thereby increase target gene replacement frequency.In this study,Cfku70 and Cfku80 genes were identified from the C.fructicola genome,and the impacts of their inactivation on fungal biology and gene deletion efficiency were characterized.Deleting Cfku70or Cfku80 did not affect fungal colony morphology,vegetative growth,sporulation,conidial germination,appressorium development and pathogenicity;in the meanwhile,Cfku70 gene deletion considerably improved deletion efficiencies of three tested genes.Our results demonstrate the usefulness ofΔCfku70 as a chassis strain enabling high-efficient target gene knock-out,and lay an important foundation for identifying novel virulence factors through the large scale gene deletion strategy in C.fructicola.

关 键 词：果生刺盘孢 同源重组 基因敲除 致病 

分 类 号：S432.44[农业科学—植物病理学]