真菌病毒BmPV1抑制Bipolaris maydis侵染寄主作用研究  

Inhibitory effects of mycoviruse BmPV1 on Bipolaris maydis infection

作　　者：王亚娇[1] 及增发[2] 栗秋生[1] 吴玉星 韩森 孔令晓[1] WANG Yajiao;JI Zengfa;LI Qiusheng;WU Yuxing;HAN Sen;KONG Lingxiao(Plant Protection Institute,Hebei Academy of Agriculture and Forestry Sciences,Key Laboratory of Integrated Pest Management on Crops in Northern Region of North China,Ministry of Agriculture and Rural Affairs,P.R.China,IPM Innovation Center of Hebei Province,International Science and Technology Joint Research Center on IPM of Hebei Province,Baoding 071000,Hebei,China;Hebei Academy of Agriculture and Forestry Sciences,Shijiazhuang 050000,Hebei,China)

机构地区：[1]河北省农林科学院植物保护研究所、农业农村部华北北部作物有害生物综合治理重点实验室、河北省农业有害生物综合防治技术创新中心、河北省作物有害生物综合防治国际科技联合研究中心,河北保定071000 [2]河北省农林科学院,河北石家庄050000

出　　处：《菌物学报》2024年第6期96-104,共9页Mycosystema

基　　金：河北省自然科学基金(C2021301033,C2023204127)。

摘　　要：玉米小斑病是我国玉米生产上的重要病害之一,每年造成玉米大幅减产。Bipolarismaydis partitivirus 1(BmPV1)病毒是玉米小斑病菌B.maydis携带的一种dsRNA病毒,具有弱毒性,能够引起B.maydis的致病力降低,但BmPV1对寄主侵染的影响并不明确。本研究采用农杆菌转化法将gfp基因分别导入到携带BmPV1病毒的玉米小斑病菌株3086-BmPV1以及脱毒菌株3086-non中,通过PCR扩增及荧光观察分析荧光标记菌株的稳定性;检测gfp基因的插入对菌株生物学特征的影响;采用叶面喷施孢子悬浮液的方式接种,并利用共聚焦显微镜观察带毒菌株及脱毒菌株对玉米叶片的侵染情况。PCR扩增和荧光观察结果表明,gfp基因成功整合到菌株3086-BmPV1和3086-non中;生物学特征分析表明gfp基因的插入对菌株的生长速度、产孢量和致病力无显著影响;侵染观察发现脱毒菌株3086-non-GFP对玉米叶片侵染有两条途径:直接侵染玉米表皮细胞并向邻近细胞扩散;侵染气孔,并通过气孔沿着细胞间隙向邻近细胞扩散。带毒菌株3086-BmPV1-GFP只能在叶片表面生长,多数不能侵染表皮细胞。综上所述,本研究观察了玉米小斑病菌带毒和不带毒菌株对玉米的侵染过程,明确了病毒BmPV1能够通过抑制小斑病菌对玉米叶片的侵入达到弱毒作用。Corn southern leaf blight is one of the important diseases in maize production in China,causing significant yield reduction every year.Bipolaris maydis partitivirus 1(BmPV1)is a dsRNA virus carried by B.maydis,the pathogens of corn southern leaf blight.It has hypovirulence and can reduce the pathogenicity of B.maydis,but the effect of BmPV1 on host infection is not clear.In this study,the Agrobacterium-mediated transformation method was used to introduce the gfp gene into the BmPV1 carrying strain 3086-BmPV1 and the non-virus strain 3086-non of B.maydis.The stability of the fluorescent-labeled strains was analyzed by PCR and fluorescence observation.The effect of the introduction of gfp gene on biological characteristics of the fluorescent-labeled strains were analyzed.Spore suspensions were sprayed on maize leaves for inoculation,and the infection of B.maydis on maize leaves was observed using fluorescence microscopy.PCR amplification and fluorescence observation results showed that the gfp gene was successfully integrated into strains 3086-BmPV1 and 3086-non.There were no significant differences in growth rate,sporulation,and pathogenicity between the fluorescent-labeled strains and the non-labeled strains.The infection results revealed that the non-virus strain 3086-non-GFP could invade maize leaves through two pathways:directly infecting epidermal cells and spreading to adjacent cells,or invading stomata and spreading through intercellular spaces.The virus-carrying strain 3086-BmPV1-GFP could only grow on the leaf surface and had minimal ability to infect epidermal cells.This study clarifies that the BmPV1 virus can reduce the pathogenicity and inhibit the infection of B.maydis.

关 键 词：玉米小斑病 真菌病毒BmPV1 弱毒性 荧光标记 侵染过程 

分 类 号：S435.131.4[农业科学—农业昆虫与害虫防治]