| 基于RAPD分子标记的烟草青枯病菌特异引物筛选及效果评价 |
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| 引用本文： | 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.基于RAPD分子标记的烟草青枯病菌特异引物筛选及效果评价[J].中国烟草学报,2021,27(2):72-78. |
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| 作者姓名： | 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 |
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| 作者单位： | 1.烟草行业黄淮烟区烟草病虫害绿色防控重点实验室 河南省农业科学院烟草研究所, 许昌 461000 |
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| 基金项目： | 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 |
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| 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...
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| 关 键 词： | RAPD 烟草青枯病 特异引物 分子检测 |
| 收稿时间： | 2020-09-15 |
Screening and evaluation of specific primers for Ralstonia solanacearum based on RAPD Technology |
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| Affiliation: | 1.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, China2.Zhumadian Municipal Tobacco Company, Zhumadian 463000, Henan, China3.Henan Provincial Tobacco Company, Zhengzhou 450018, China |
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| Abstract: | 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 6×102 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. |
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