李树炭疽病病原果生刺盘孢特异性PCR检测  

Specific PCR detection of Colletotrichum fructicola causing anthracnose on plum trees

作　　者：陈小林[1] 鲁萌萌 唐利华[1] 黄穗萍[1] 郭堂勋[1] 李其利[1] CHEN Xiaolin;LU Mengmeng;TANG Lihua;HUANG Suiping;GUO Tangxun;LI Qili(Institute of Plant Protection,Guangxi Academy of Agricultural Sciences/Key Laboratory of Green Prevention and Control on Fruits and Vegetables in South China Ministry of Agriculture and Rural Affairs/Guangxi Key Laboratory of Biology for Crop Diseases and Insect Pests,Nanning 530007,China)

机构地区：[1]广西壮族自治区农业科学院植物保护研究所,农业农村部华南果蔬绿色防控重点实验室,广西作物病虫害生物学重点实验室,南宁530007

出　　处：《植物病理学报》2024年第5期1020-1028,共9页Acta Phytopathologica Sinica

基　　金：广西重点研发计划(桂科AB20159041)。

摘　　要：果生刺盘孢(Colletotrichum fructicola)是引起李树炭疽病的优势病原菌,属于胶孢刺盘孢复合群(C.gloeosporioides species complex)。本研究对该复合群中36个菌株的ApMat序列进行比对,进而根据C.fructicola特异性位点设计了1对针对该菌的特异性引物:F2(5′-CGTGACCCAGGAGGCGACCACGCATCTGT-3′)和R2(5′-GGTGATCTCCTAGCGTTCGTACGTCTAAT-3′),在此基础之上建立了PCR检测体系。结果表明,引物F2;R2可从C.fructicola基因组DNA中扩增出长度为132 bp的特异性目的条带,检测灵敏度达100 fg·μL^(-1)。利用该检测体系可从人工接种C.fructicola HN47-2菌株的李树叶片以及田间自然染病的李树叶片样品中快速检测出目标条带。本研究针对C.fructicola建立的灵敏、快速的特异性PCR检测方法为该菌引起的李树炭疽病的田间早期诊断、动态监测和精准防控提供了重要的技术支撑。Colletotrichum fructicola,belonging to the C.gloeosporioides species complex,is the dominant pathogenic species causing anthracnose on plum trees.In this study,multiple alignment of ApMat sequences of 36 strains in the C.gloeosporioides complex was performed,a pair of C.fructicola-specific primers:F2(5′-CGTGACCCAGGAGGCGACCACGCATCTGT-3′)and R2(5′-GGTGATCTCTCCTAGCGTTCGTACGTCTAAT-3′)was designed based on the specific sequences of the pathogen,and a PCR detection method was develo-ped accordingly.The results showed that a specific target amplicon of 132 bp could only be produced by C.fructicola with the primer pair F2;R2,with a detection sensitivity of 100 fg·μL^(-1).Using this PCR detection system,the target pathogen C.fructicola can be quickly detected from the leaves of plum trees artificially inoculated with C.fructicola HN47-2 strain and the leaves of plum trees naturally infected with C.fructicola.The C.fructicola-specific PCR detection method established in this study provides an important technical support for early diagnosis,dynamic monitoring and accurate prevention and control of plum anthracnose caused by C.fructicola in the field.

关 键 词：李树炭疽病 果生刺盘孢 特异性引物 PCR检测 

分 类 号：S432.1[农业科学—植物病理学]