Rapid diagnosis ofRalstonia solanacearuminfection sweet potato in China by loop-mediated isothermal amplification

文献类型： 外文期刊

第一作者： Li, Huawei

作者： Li, Huawei;Zhang, Hong;Liu, Zhonghua;Lin, Zhijian;Qiu, Yongxiang;Tang, Hao;Qiu, Sixin;Li, Huawei;Zhang, Hong;Liu, Zhonghua;Lin, Zhijian;Qiu, Yongxiang;Tang, Hao;Qiu, Sixin

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关键词： Sweet potato; Bacterial wilt; R; solanacearum; LAMP

期刊名称：ARCHIVES OF MICROBIOLOGY （ 影响因子：2.552； 五年影响因子：2.475 ）

ISSN： 0302-8933

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收录情况： SCI

摘要： Bacterial wilt of sweet potato is caused byRalstonia solanacearum, which is distributed in southern China and causes significant economic losses each year. The pathogen is soil- and rhizome-borne, and thus its rapid detection may prevent the occurrence and spread of the disease.R. solanacearumhas been listed as a quarantine disease in China. With the advent of molecular biology, many novel tools have been explored for the rapid identification of plant pathogens. In this study, a strain-specific detection method was developed for this specific pathogen that infects sweet potato using loop-mediated isothermal amplification (LAMP). A set of new LAMP-specific primers was designed from theorf428gene, which can specifically detect theR. solanacearumbacterium that infect sweet potato. The LAMP reaction consisted of 8.0 mmol center dot L-1Mg2+, 1.4 mmol center dot L(-1)dNTPs, and 0.32U mu L(-1)Bst 2.0 DNA polymerase and was performed at 65 degrees C for 1 h. The amplification products were detected by visualizing a mixture of color changes using SYBR Green I dye and assessing ladder-like bands by electrophoresis. Our method has specificity, i.e., it only detectedR. solanacearumin sweet potato, and it has high sensitivity, with a detection limit of 100 fg center dot mu L(-1)genomic DNA and 10(3) CFU center dot mL(-1)of bacterial fluid. In addition,R. solanacearumcould be directly detected in infected sweet potato tissues without the need for DNA extraction. The LAMP method established in this study is a highly specific, sensitive, and rapid tool for the detection of bacterial wilt in sweet potato caused byR. solanacearum.

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