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

文献类型： 外文期刊

作者： Li, Huawei 1 ; Zhang, Hong 1 ; Liu, Zhonghua 1 ; Lin, Zhijian 1 ; Qiu, Yongxiang 1 ; Tang, Hao 1 ; Qiu, Sixin 1 ;

作者机构： 1.Fujian Acad Agr Sci, Inst Crop Sci, Fuzhou 350013, Fujian, Peoples R China

2.Minist Agr, Sci Observing & Expt Stn Tuber & Root Crops South, Fuzhou 350013, Fujian, Peoples R China

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

年卷期：

页码：

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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