基于多重PCR技术的西瓜噬酸菌分组检测方法及其应用  

Development and application of a multiplex PCR assay for grouping of Acidovorax citrulli

作　　者：杨玉文[1] 乔培 刘德华 费诺亚 关巍 赵廷昌[1] YANG Yuwen;QIAO Pei;LIU Dehua;FEI Nuoya;GUAN Wei;ZHAO Tingchang(State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Jilin Agricultural University,Changchun 130118,China;Shenyang Agricultural University,Shenyang 110866,China)

机构地区：[1]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193 [2]吉林农业大学,长春130118 [3]沈阳农业大学,沈阳110866

出　　处：《植物保护》2022年第4期211-219,共9页Plant Protection

基　　金：国家西甜瓜产业技术体系(CARS-25);国家重点研发计划(2018YFD0201304);中国农业科学院科技创新工程(CAAS-ASTIP)。

摘　　要：瓜类细菌性果斑病是瓜类作物上重要的种传细菌性病害,其病原菌西瓜噬酸菌Acidovorax citrulli为我国进境植物检疫性有害生物,种子种苗带菌是病害长距离传播的重要来源,种子种苗的快速检测对病害综合防控具有重要的意义。根据寄主的差异西瓜噬酸菌分为两个组,Ⅱ组菌株比Ⅰ组菌株具有更高的铜制剂敏感性,因此,西瓜噬酸菌的分组检测可为病害田间防治中铜制剂的精准使用提供科学依据,避免化学农药的过量施用。本研究筛选了现有的西瓜噬酸菌种间特异性引物和种内分组特异引物,建立并优化了一个多重PCR体系,实现了通过一步试验,就能够准确将西瓜噬酸菌与近缘种和其他植物病原细菌区分开来,并直接鉴定到西瓜噬酸菌不同组。该多重体系可以从带菌种子浸泡液和感病植物组织研磨液中直接检测到西瓜噬酸菌的不同组,且稳定性好,具有应用于生产实践的潜力。Bacterial fruit blotch(BFB)is an important seed-borne bacterial disease of cucurbits.Its pathogen,Acidovorax citrulli,is one of quarantine pests at points of entry in China.Seeds and seedlings are identified as the most primary inoculum source in long-distance transmission.Rapid detection of seeds and seedlings is critical for disease management.A.citrulli can be divided into two groups(ⅠandⅡ)with varying host ranges,and groupⅡstrains are more sensitive to copper than groupⅠones.Therefore,accurately distinguishing A.citrulli groups could improve the efficacy of copper-based compounds for BFB management in the field.In this study,based on the existing A.citrulli species-specific primers and specific primers for intraspecific grouping,we designed and optimized a multiplex PCR assay,which could be used to distinguish the two groups of A.citrulli from other plant pathogenic bacteria in a single reaction.The multiplex PCR assay was assessed by testing the seed washings from the bacteria-carrying seeds and the grinding fluid of diseased plant tissues.These results suggest that it is accurate and stable,and has the potential for application in production.

关 键 词：西瓜噬酸菌 检测 多重PCR 分组 

分 类 号：S432.42[农业科学—植物病理学]