根部内生真菌—寄生疫霉—拟南芥互作模式体系的构建与鉴定  

Construction and Identification of System for Root Endophyte-Phytophthora parasitica-Arabidopsis thaliana Interaction Model

作　　者：金明 张彩虹 董晨曦 史青尧 张飞[2] 单卫星[1] 强晓玉 JIN Ming;ZHANG Caihong;DONG Chenxi;SHI Qingyao;ZHANG Fei;SHAN Weixing;QIANG Xiaoyu(College of Agronomy,Northwest A&F University,Yangling Shaanxi 712100,China;College of Horticulture,Northwest A&F University,Yangling Shaanxi 712100,China)

机构地区：[1]西北农林科技大学农学院,陕西杨凌712100 [2]西北农林科技大学园艺学院,陕西杨凌712100

出　　处：《西北农业学报》2023年第10期1656-1664,共9页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家重点研发计划(2017YFD0200602-2);国家自然科学基金(31872657);国家现代农业产业技术体系(CARS-09)。

摘　　要：为构建根部内生真菌—寄生疫霉—拟南芥三方互作模式研究体系,以影响马铃薯晚疫病害发生的潜在内生真菌雪球微座孢(Microdochium bolleyi)为例,利用WGA-Alexa Fluor■488染色和带有GFP标记的寄生疫霉(Phytophthora parasitica)转化菌株,通过荧光显微镜观察分析M.bolleyi侵染拟南芥根部以及M.bolleyi与寄生疫霉和拟南芥根部三方互作的细胞学特征;利用皿内平板拮抗测试初步分析内生真菌M.bolleyi对致病疫霉(Phytophthora infestans)的抑制效果,并进一步利用皿内拟南芥与真菌M.bolleyi和寄生疫霉三方互作体系,鉴定分析真菌M.bolleyi对植物与寄生疫霉亲和互作的影响。结果表明,真菌M.bolleyi可成功侵染并定殖于根组织,而未引发植物细胞坏死现象,说明M.bolleyi具有典型的内生真菌特征;相比于对照预处理,真菌M.bolleyi预处理能够抑制寄生疫霉在拟南芥根部的定殖和扩展,初步揭示真菌M.bolleyi在植物与疫霉菌互作中的影响作用;真菌M.bolleyi不仅对致病疫霉的皿内抑制率可达63.55%,还能够显著减轻拟南芥根部对寄生疫霉的感病性。可见,所建立的根部内生真菌—寄生疫霉—拟南芥三方互作模式体系不仅为快速鉴定有益真菌影响植物抗病抗逆生物学功能提供了技术体系,也为深入解析植物根部与不同微生物之间的互作机制奠定基础。To construct the tripartite interaction model research system of“Root endophyte-Phytophthora parasitica-Arabidopsis thaliana”,the fungus Microdochium bolleyi which has potential effect on the potato late blight diseasewas taken asan example,with the application of either WGA-Alexa Fluor■488 staining or GFP-tagged P.parasitica transformant,cytological characteristics of A.thaliana Col-0 roots with either colonization of M.bolleyi or inoculation of both M.bolleyi and P.parasitica were analyzed by using fluorescence microscopy.By using the established tripartite interaction system of Arabidopsis root with M.bolleyi and P.parasitica,the effect of M.bolleyi on the compatible interaction between Arabidopsis and P.parasitica was analyzed.The results showed that M.bolleyi could successfully penetrate and colonize Arabidopsis root with no occurrence of root cell disruption,indicating that M.bolleyi maintains the typical features of an endophyte.Besides,the pre-treatment of M.bolleyi rather than mock solution could inhibit the expansion and colonization of P.parasitica in A.thaliana roots,indicating that M.bolleyi could affect the plant-Phytophthora interaction.Further antagonism test indicated M.bolleyi could inhibit the growth of P.infestans,with an inhibitory rate of 63.55%,and the pre-treatment of M.bolleyi could significantly attenuate the Arabidopsis root susceptibility to P.parasitica.Taken together,the established tripartite interaction system with root endophyte,P.parasitica and Arabidopsis root could not only provide a fast identification tool for biological function of candidate endophytes,but also lays a solid foundation for mechanistic analysis of plant-pathogen-beneficial microbe interactions.

关 键 词：马铃薯晚疫病 内生真菌 寄生疫霉 拟南芥 雪球微座孢 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]