摘要
为实现植物青枯病的早期诊断,需要建立一种适于田间快速便捷检测青枯病菌的方法。以细胞色素C基因为靶标设计一套特异性引物,建立了植物青枯病的LAMP检测方法。此方法最低检测极限为1 pg,可在1 h内完成,不依赖昂贵复杂的仪器,结果可经肉眼观察。利用此方法,在人工接种发病的茄子、番茄、花生、芝麻和凹头苋茎部浸出液和马铃薯病薯块茎组织液中均检测出青枯病菌的存在,尤其适用于田间疑似罹病的芝麻、花生、番茄、马铃薯和甘薯等植株的检测,且LAMP法的检出率远高于PCR法。应用LAMP技术检测青枯病菌快速高效、特异性强、灵敏度高,操作简单,适于在基层推广运用。
The purpose of the study is to establish a rapid detection method for Ralstonia solanacearum,which is beneficial to the early diagnosis of plant bacterial wilt in the field.The LAMP(loop-mediated isothermal amplification)method for amplification of cytochrome C gene was established by optimization of reaction conditions.The minimum detection limit with the LAMP method reached 1 pg.The detection could be completed within 1 h without using complicated instruments and the results were visible by human eye.Using this method,R.solanacearum was detected in the stem extracts of manually inoculated eggplant,tomato,peanut,sesame and Amaranthus lividus and potato tuber tissue fluid of potato,especially,it was appropriate for the detection of sesame,peanut,tomato,potato and sweet potato plants suspected of being in disease in the field,and the detection rate by LAMP was much higher than that by PCR method.The LAMP method is a fast,specific,sensitive,and easy-to-use for the field detection of R.solanacearum bacterial wilt,which is suitable for promotion at the grassroots level.
作者
李信申
黄小梅
吴淑秀
黄瑞荣
魏林根
华菊玲
LI Xin-shen;HUANG Xiao-mei;WU Shu-xiu;HUANG Rui-rong;WEI Lin-gen;HUA Ju-ling(Plant Protection Institute,Jiangxi Academy of Agricultural Sciences,Nanchang 330200;Jiangxi Vocational College of Science and Technology,Nanchang 330200;Yingtan Agricultural and Grain Bureau,Yingtan 335000;Soil and Fertilizer and Resources and Environment Institute,Jiangxi Academy of Agricultural Sciences,Nanchang 330200)
出处
《生物技术通报》
CAS
CSCD
北大核心
2021年第1期272-281,共10页
Biotechnology Bulletin
基金
国家芝麻产业体系建设项目(CARS-15-2-05)
江西省薯类产业体系建设项目(JXARS-19-03)
国家自然科学基金项目(31360428)
江西省农业科学院博士启动项目(20142C13S006)。
关键词
青枯雷尔氏菌
环介导等温扩增
快速检测
Ralstonia solanacearum
loop-mediated isothermal amplification
rapid detection
作者简介
李信申,男,博士,研究方向:植物细菌病害防控,E-mail:lixinshen0@163.com;通讯作者:华菊玲,女,研究员,研究方向:植物保护,E-mail:huajl2000@126.com。
