Real-time Fluorescence PCR Method for Detection of Burkholderia glumae from Rice
Real-time Fluorescence PCR Method for Detection of Burkholderia glumae from Rice
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DOI：
10.1016/s1672-6308(08)60073-6
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发表时间：
2009-06-01
期刊：
影响因子：
4.8
通讯作者：
Xie Guan-Lin
中科院分区：
文献类型：
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作者：
Fang Yuan;Xu Li-hui;Xie Guan-Lin
2007年，中国将引起水稻苗腐病和谷粒腐病的葡萄伯克霍尔德菌列为植物检疫病害。建立有效的病原菌检测方法来控制该病的进一步传播是十分必要的。本研究将实时PCR方法与经典PCR方法相结合，提高检测效率，建立一种准确、快速、灵敏的种子检疫病原菌检测方法，为病害的有效管理提供依据。结果表明，所有菌株的实时PCR和普通PCR均产生了约139bp的特异性片段，其余菌株的PCR结果均为阴性。一般PCR法在浓度为1 × 10(4) CFU/mL时检测到细菌，实时荧光PCR法在浓度低于100 CFU/mL时检测到细菌。接种后的种子与健康种子按1:100的比例混合，可检出B.glumae。
Burkholderia glumae causing seedling rot and grain rot of rice was listed as a plant quarantine disease of China in 2007. It's quite necessary to set up effective detection methods for the pathogen to manage further dispersal of this disease. The present study combined the real-time PCR method with classical PCR to increase the detecting efficiency, and to develop an accurate, rapid and sensitive method to detect the pathogen in the seed quarantine for effective management of the disease. The results showed that all the tested strains of B.glumae produced about 139 bp specific fragments by the real-time PCR and the general PCR methods, while others showed negative PCR result. The bacteria could be detected at the concentrations of 1x10(4) CFU/mL by general PCR method and at the concentrations below 100 CFU/mL by real-time fluorescence PCR method. B.glumae could be detected when the inoculated and healthy seeds were mixed with a proportion of 1:100.