假禾谷镰孢菌弱毒菌株FC136-2A的GFP标记转化子构建  

Construction of GFP-labeled Transformant of Hypovirulent Strain FC136-2A in Fusarium pseudograminearum

作　　者：王志芳 李轲 刘东伟 谢源 代君丽[1] 王珂 高飞[1] 张晓婷[1] 李洪连[1,2] WANG Zhi-Fang;LI Ke;LIU Dong-Wei;XIE Yuan;DAI Jun-Li;WANG Ke;GAO Fei;ZHANG Xiao-Ting;LI Hong-Lian(College of Plant Protection,Henan Agricultural University,Zhengzhou 450002,China;National Key Laboratory of Wheat and Maize Crop Science,Zhengzhou 450046,China)

机构地区：[1]河南农业大学植物保护学院,郑州450002 [2]省部共建小麦玉米作物学国家重点实验室,郑州450046

出　　处：《农业生物技术学报》2021年第7期1435-1442,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金国际(地区)合作与交流项目(31961143018);河南省科技攻关计划(农业领域)项目(212102110138);2018年度河南省高等学校青年骨干教师培养计划(2018GGJS032);河南农业大学科技创新基金(KJCX2020A14)。

摘　　要：茎基腐病是在世界上多数小麦(Triticum aestivum)种植区广泛发生的重要土传病害,假禾谷镰孢菌(Fusarium pseudograminearum)是该病的优势病原菌。假禾谷镰孢菌巨型双链RNA病毒1(Fusarium pseudograminearum megabirnavirus 1,FpgMBV1)是在假禾谷镰孢菌菌株FC136-2A中发现的真菌病毒,使该菌株在小麦上的致病力显著降低,即具有弱毒作用。为了研究FpgMBV1的弱毒作用机制,本研究利用农杆菌(Agrobacterium tumefaciens)介导的遗传转化技术将pCAM-GFP-hyg导入FC136-2A菌株,并对转化子FC136-2A-GFP进行分子检测、荧光观察、生物学鉴定和致病力测定。结果表明,带有绿色荧光标记的转化子FC136-2A-GFP与FC136-2A在生长速率、产孢量和对小麦的致病力等生物学性状上差异不显著,且转化子FC136-2A-GFP仍然带有FpgMBV1。本研究获得带有GFP标记的假禾谷镰孢菌转化子FC136-2A-GFP,可用于观察弱毒菌株FC136-2A在小麦上的侵染和致病过程,为深入开展真菌病毒FpgMBV1对假禾谷镰孢菌的弱毒作用机制研究提供基础资料。Wheat crown rot is an important soil-borne disease occurring widely in most wheat(Triticum aestivum) growing areas in the world. Fusarium pseudograminearum is the dominant causal agent of wheat crown rot. In F. pseudograminearum strain FC136-2 A, mycovirus Fusarium pseudograminearum megabirnavirus 1(FpgMBV1) was found to cause the hypovirulence of FC136-2 A on wheat. To explore the mechanism of hypovirulence caused by FpgMBV1, a pCAM-GFP-hyg construct was transformed into F.pseudograminearum strain FC136-2 A by Agrobacterium tumefaciens-mediated genetic transformation technology. Molecular identification, fluorescent observation, biological characterization and pathogenicity test of the transformants were also conducted. Results showed that the GFP-labeled transformant FC136-2 AGFP had no biological differences compared to FC136-2 A in growth rate and sporulation capacity. Besides,FpgMBV1 was still in the transformant FC136-2 A-GFP and the pathogenicity of FC136-2 A-GFP was low on wheat. The present study obtained GFP-labeled transformant FC136-2 A-GFP of F. pseudograminearum, which could be used to observe the infection and pathogenic process of hypovirulence strain FC136-2 A on wheat,and provide basic information for further study on the mechanism of hypovirulence effect of mycovirus FpgMBV1 on F. pseudograminearum.

关 键 词：小麦茎基腐病 假禾谷镰孢菌 真菌病毒 假禾谷镰孢菌巨型双链RNA病毒1(FpgMBV1) 弱毒作用 

分 类 号：S432.23[农业科学—植物病理学]