A LFD-RAA-Based Rapid Detection Method for Pseudomonas syringae pv. tabaci  

烟草野火病菌LFD-RAA快速检测方法的建立

作　　者：XIAO Yan-song LI Hong-guang LI Si-jun WU Wen-xin ZHOU Lu-su ZHONG Jie SU Jia-en YANG Zhi-juan 肖艳松;李宏光;李思军;吴文信;周陆苏;钟杰;苏家恩;杨志娟(湖南省烟草公司郴州市公司,湖南郴州423000;湖南省烟草公司郴州市公司桂阳县公司,湖南郴州424400;湖南农业大学植物保护学院,湖南长沙410128;云南省烟草公司大理市公司,云南大理671000)

机构地区：[1]Chenzhou Tobacco Company,Hunan Provincial Tobacco Corporation,Chenzhou 423000,PRC [2]Guiyang Branch of Chenzhou Tobacco Company,Hunan Provincial Tobacco Corporation,Chenzhou 424400,PRC [3]College of Plant Protection,Hunan Agricultural University,Changsha 410128,PRC [4]Dali Tobacco Company,Yunnan Provincial Tobacco Corporation,Dali 671000,PRC

出　　处：《Agricultural Science & Technology》2024年第2期38-43,共6页农业科学与技术（英文版）

基　　金：湖南省郴州市烟草公司烟草农业科技创新项目(CZYC2022JS03,CZYC2023JS05);云南省烟草公司大理州公司科技计划一般项目(DLYC2022003)。

摘　　要：This study aimed to achieve rapid detection of Pseudomonas syringae pv.tabaci,the pathogen of tobacco wildfire disease.The specific primers and probes for recombinase-aided amplification(RAA)were designed with HrpZ as the target gene.RAA was then combined with the lateral flow dipstick(LFD)to establish a LFD-RAA-based rapid detection system for the pathogen.Furthermore,the detection performance of the established method was tested.The results showed that the LFDRAA method had high specificity.The amplification could be completed after 25 min of reaction at 39℃.The sensitivity of the established method reached 0.0001 ng/μL,which was superior to that of PCR detection.Moreover,the LFD-RAA method could quickly detect P.syringae pv.tabaci from tobacco leaves,demonstrating field applicability.To sum up,the LFD-RAA method established in this study can be applied in the rapid detection and early diagnosis of tobacco wildfire disease.为快速有效检测烟草野火病菌(Pseudomonas syringae pv.tabaci),以烟草野火病菌HrpZ基因序列为靶标,设计重组酶介导扩增技术(recombinase aided amplification,RAA)特异性引物,结合侧流层析试纸条,获得了特异性的引物和探针,建立了烟草野火病菌LFD-RAA快速检测体系,并对其检测能力进行验证。结果表明:该检测体系具有较高的特异性,在39℃恒温条件下反应25 min即可完成扩增,灵敏度可达到0.0001 ng/μL,优于PCR检测灵敏度,且该检测体系可从烟草叶片上快速检测到烟草野火病菌,证明其实用性强。综上所述,LFD-RAA检测体系可应用于烟草野火病的快速检测和早期诊断。

关 键 词：Tobacco wildfire disease Recombinase-aided amplification DETECTION 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]