转红色荧光蛋白稻瘟菌的构建与分子分析  

Construction and molecular analysis of Magnaporthe oryzae transformed with red fluorescent protein

作　　者：田旭丹 李亚丽[2] 应淑敏 李利华[4] 李莘 王岚岚 龙文莉[2] 王教瑜[3] 瞿绍洪[2] TIAN Xudan;LI Yali;YING Shumin;LI Lihua;LI Xin;WANG Lanlan;LONG Wenli;WANG Jiaoyu;QU Shaohong(College of Chemistry and Life Sciences,Zhejiang Normal University,Jinhua,Zhejiang 321004,China;Institute of Virology and Biotechnology,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;Institute of Plant Protection and Microbiology,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;College of Plant Protection,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]浙江师范大学化学与生命科学学院,浙江金华321004 [2]浙江省农业科学院病毒学与生物技术研究所,杭州310021 [3]浙江省农业科学院植物保护与微生物研究所,杭州310021 [4]湖南农业大学植物保护学院,长沙410128

出　　处：《植物生理学报》2022年第5期972-980,共9页Plant Physiology Journal

基　　金：国家自然科学基金(31672016);浙江省农业科学院省部共建国家重点实验室自主设计课题。

摘　　要：水稻稻瘟病是水稻重要病害之一,研究稻瘟菌与水稻互作,对水稻稻瘟病防治具有重要意义。通过原生质体转化法将红色荧光蛋白基因RFP转化稻瘟菌菌株E2007046A-2,获得了6个稳定的转化子;通过继代培养,对转化子进行表型鉴定、DNA杂交、qRT-PCR分析,筛选出了产孢量较高、RFP和HPT稳定插入且高表达的转化菌株;在荧光显微镜下,观察到水稻细胞内表达红色荧光蛋白的侵染菌丝形成的侵染结构。本研究建立了稻瘟菌侵染水稻过程的可视化系统,为稻瘟菌与水稻互作提供了材料和研究基础。Rice blast is one of the important diseases of rice.The study on the interaction between Magnaporthe oryzae and rice is of great significance for the control of rice blast.In this study,the red fluorescent protein(RFP)gene was transformed into M.oryzae strain E2007046A-2 by protoplast transformation,and six stable transformants were selected.Transgenic M.oryzae strains derived from multi-generation subculture were characterized by phenotypic observation,DNA hybridization and qRT-PCR,from which high spore-output transformants with stable insertion and high expression of HPT and RFP were obtained.Under the fluorescence microscope,the infection structure in rice cell formed by M.oryzae fungus hyphae with stable RFP expression was observed.This study establishes a research system visualizing the process for M.oryzae to infect the rice host and provides a basis for the study of the M.oryzae-rice interaction.

关 键 词：稻瘟菌 红色荧光蛋白 水稻 遗传转化 

分 类 号：S435.111.41[农业科学—农业昆虫与害虫防治]