摘要
为建立烟田土壤和烟株烟草青枯病菌的快速诊断方法,本研究采用RAPD方法筛选获得了6对可用于烟草青枯病菌PCR检测的特异性引物。经对6种细菌(烟草青枯病菌、多粘芽孢杆菌、枯草芽孢杆菌、节杆菌、荧光假单胞菌、野火病菌)和2个烟草品种(中烟100和云烟87)的检测验证,6对引物均具有良好的灵敏度、特异性和可靠性。对青枯菌菌液DNA的检测灵敏度可达0.42 pg/μL,对土壤青枯菌的最低检出率为6×10^(2) cfu/g。应用特异引物A1T和S13T可对接种土壤和烟株样品的研磨悬浮液直接进行PCR快速检测,显著地缩短了检测时间,对烟叶产区烟草青枯病的预测预报及防控具有重要意义。
In order to realize rapid diagnosis of Ralstonia solanacearum in the tobacco plant and tobacco field soil,six pairs of specific primers for PCR detection of tobacco bacterial wilt were obtained by RAPD method in this study.According to the tests by six kinds of bacteria (R.solanacearum,Bacillus polymyxa,Bacillus subtilis,Arthrobacter,Pseudomonas fluorescens,Pseudomonas tabaci) and two tobacco varieties (Zhongyan 100 and Yunyan 87),the 6 pairs of specific primers showed good sensitivity,specificity and reliability.The detection sensitivity of DNA in bacterial liquor was 0.42 pg/uL,and the minimum detection rate of soil bacterial wilt was 6×10^(2) cfu/g.The results of PCR amplification using specific primers A1T and S13T showed that the application of two primers realized rapid detection of Ralstonia solanacearum in tobacco planting soil and infected tobacco plants,so that the detetion time was shortened significantly.The results of this study are of great significance for the forecasting and preventing of tobacco bacterial wilt.
作者
李小杰
刘畅
李成军
杨立均
邱睿
宋瑞芳
陈玉国
白静科
李淑君
LI Xiaojie;LIU Chang;LI Chengjun;YANG Lijun;QIU Rui;SONG Ruifang;CHEN Yuguo;BAI Jingke;LI Shujun(Key Laboratory for Green Preservation&Control of Tobacco Diseases and Pests in Huanghuai Growing Area Tobacco Research Institute of Henan Academy of Agricultural Sciences,Xuchang 461000,China;Zhumadian Municipal Tobacco Company,Zhumadian 463000,Henan,China;Henan Provincial Tobacco Company,Zhengzhou 450018,China)
出处
《中国烟草学报》
CAS
CSCD
北大核心
2021年第2期72-78,共7页
Acta Tabacaria Sinica
基金
中国烟草总公司重点项目“河南烟草病虫害绿色防控关键技术研究与应用”(201741000027063)
河南省烟草公司科技项目“河南省烟草主要病虫害预测预报及综合防治示范”(201741000027064)
驻马店市烟草公司科研项目“烟草主要病害的“烟株-土壤”双效生防技术研究与应用”(201641170024101)。
关键词
RAPD
烟草青枯病
特异引物
分子检测
RAPD
tobacco bacterial wilt
specific primers
molecular detection
