油茶(Camellia oleifera)果生刺盘孢菌(Colletotrichum fructicola)Hog1基因功能和致病性分析  

Analysis on the Function and Pathopoiesia of Camellia oleifera Colletotrichum fructicola Hog1 Gene

作　　者：刘慧年 周国英[1] 刘君昂[1] Liu Huinian;Zhou Guoying;Liu Junang(Hunan Provincial Key Laboratory ofControl of Harmful Forest Living Beings,Key Laboratory of Cultivation and Protection for Non-wood Forest Trees,Ministry of Education,Central South University ofForestry and Technology,Changsha,410004)

机构地区：[1]中南林业科技大学,森林有害生物防控湖南省重点实验室,经济林培育与保护教育部重点实验室,长沙410004

出　　处：《分子植物育种》2020年第11期3517-3523,共7页Molecular Plant Breeding

基　　金：国家重点研发计划课题(2017YFD0600103);湖南省研究生科技创新基金(CX2018 B456);中南林业科技大学研究生科技创新基金(20183001)共同资助。

摘　　要：为了探讨Hog1基因在油茶果生刺盘孢菌(Colletotrichum fructicola)致病过程中的作用。本研究以酿酒酵母中的Hog1基因为依托,进行BlAsTP比对分析油茶果生刺盘孢菌的全基因组数据,以获取与其高度同源的Hog1蛋白序列,通过对ATMT体系进行改进和调整,建立了炭疽病菌基因敲除体系。采用形态观察、显微鉴定、生物学表型分析方法,基于科赫法则(Koch postulates),检测病原菌的致病力大小。结果表明,本试验成功敲除了C.fructicola野生型菌株CFLH16 Hog1基因,发现突变体菌株ΔC.f Hog1与CFLH16相比,其产孢量显著下降,且无法形成附着胞,基本丧失致病能力;ΔC.f Hog1对盐胁迫(NaCl)与CFLH16相比其敏感性显著降低,对细胞壁胁迫(SDS)和过氧化物胁迫(H2O2)与CFLH16相比其敏感性不明显。研究发现C.f Hog1基因编码的蛋白质是果生刺盘孢菌生长发育过程中一个重要蛋白,参与调控病菌的附着胞形成、致病过程以及对外界胁迫的应答。本试验针对油茶炭疽病害的功能基因进行探索,为油茶病害分子机制研究提供了参考依据,为进一步实现油茶病害有效调控的目标打下了基础。To examine the role of Hog1 gene in the pathogenic process of carpogenous Colletotrichum fructicola of Camellia oleifera.The highly homologous Hog1 protein sequence was acquired by conducting BlAsTP analysis of Hog1 gene in Saccharomyces cerevisiae according to the whole-genome database of carpogenous C.fructicola of Camellia oleifera.Through the improvement and adjustment of Agrobacterium tumefaciens-mediated transformation(ATMT)system,the gene knockout system of C.fructicola was established.Based on Koch postulates,the pathogenicity of pathogenic bacteria was detected;morphology observation,microscopic identification and biological phenotypic analysis were used.The sporulation yield of mutantΔC.f Hog1 significantly decreased on the Hog1 gene knockout from C.fructicola wild-type strain CFLH16,leading to the failure of appressorium formation and pathogenicity deprivation.Compared with the wild type,ΔC.f Hog1 was considerably less sensitive to NaCl stress and less significant in the sensitivity to SDS stress and H2O2 stress.It was found that the protein encoded by C.f Hog1 gene is an important protein in the growth and development of C.fruticosa,which is involved in the regulation of appressorium formation,pathogenic process and response to external stress.In this study,the functional genes of oil tea anthrax disease were explored,which provided a reference for the study of the molecular mechanism of oil tea disease,and laid a foundation for the further realization of the goal of effective control of oil tea disease.

关 键 词：油茶(Camellia oleifera) Hog1基因 果生刺盘孢菌 附着胞形成 胁迫应答 

分 类 号：S763.7[农业科学—森林保护学]