尖孢镰刀菌快速检测LAMP体系的建立及优化  

Establishment and Optimization of LAMP System for Rapid Detection ofFusarium oxysporum

作　　者：唐彤彤 李子强 林安琪 胡晓蕾 邓佳莹 孙星 TANG Tongtong;LI Ziqiang;LIN Anqi;HU Xiaolei;DENG Jiaying;SUN Xing(School of Biological and Food Engineering,Chuzhou University,Chuzhou 239000)

机构地区：[1]滁州学院生物与食品工程学院,安徽滁州239000

出　　处：《长江蔬菜》2025年第4期26-30,共5页Journal of Changjiang Vegetables

基　　金：安徽省自然科学基金(2108085MC87);安徽省高等学校科研计划项目(2022AH051093);安徽省滁菊种植与深加工工程研究中心资助项目(2024CJZX04);滁州学院大学生创新创业训练计划项目(2022CXXL102、2024CXXL111)。

摘　　要：针对当前番茄枯萎病的为害及防治难点,急需建立一种快速、准确的尖孢镰刀菌检测体系。对尖孢镰刀菌(Fusarium oxysporum)的r DNA-IGS序列设计特异性LAMP引物,初步建立LAMP快速检测体系,并对LAMP反应体系及反应条件进行优化,检测反应体系的特异性及灵敏度。同时,对以碱裂法提取DNA和以番茄不同部位(根、叶)组织液为检测模板的反应效果进行比较。结果表明,以菌株TR-4的r DNA-IGS区段为靶标设计了LAMP引物共5条;最终反应体系为dNTPs 1.0 mmol/L、Mg^(2+)2.0 mmol/L、反应温度60℃、反应时长50 min;所设计的LAMP引物可针对尖孢镰刀菌进行特异性检测,且检测灵敏度达到1 pg/μL;碱裂法提取DNA的检出效率高于番茄组织液,根的检测效率高于叶。本研究所建立的尖孢镰刀菌LAMP检测方法,可为田间番茄枯萎病的快速检验提供技术支撑。In view of the current damage and control difficulties of tomato Fusarium wilt,it is urgent to establish a rapid and accurate detection system of Fusarium oxysporum.The specific LAMP primers were designed for the rDNA-IGS sequence of F.oxysporum,and the LAMP rapid detection system was initially established.The LAMP reaction system and reaction conditions were optimized to detect the specificity and sensitivity of the reaction system.At the same time,the reaction effects of DNA extraction by alkaline cleavage method and tissue fluid of different parts(roots and leaves)of tomato as detection templates were compared.The results showed that five LAMP primers were designed based on the rDNA-IGS region of strain TR-4.The final reaction system was dNTPs 1.0 mmol/L,Mg^(2+)2.0 mmol/L,reaction temperature 60℃,reaction time 50 minutes.The designed LAMP primers could specifically detect F.oxysporum,and the detection sensitivity was 1 pg/μL.The detection efficiency of DNA extracted by alkaline lysis method was higher than that of tomato tissue fluid,and the detection efficiency of roots was higher than that of leaves.The LAMP detection method of F.oxysporum established in this study could provide technical support for the rapid detection of tomato Fusarium wilt in the field.

关 键 词：尖孢镰刀菌 番茄枯萎病 r DNA-IGS 环介导等温扩增 

分 类 号：S436.421.13[农业科学—农业昆虫与害虫防治]