Real-time Fluorescence PCR Method for Detection of Burkholderia glumae from Rice

• Published in: Rice science Vol. 16; no. 2; pp. 157 - 160
• Main Authors: Fang, Yuan, Xu, Li-hui, Tian, Wen-xiao, Huai, Yan, Yu, Shan-hong, Lou, Miao-miao, Xie, Guan-lin
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.06.2009; State Key Laboratory of Rice Biology/Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China
• Subjects: bacterial grain rot; Burkholderia glumae; detection; PCR扩增法; PCR方法; real-time fluorescence polymerase chain reaction; rice; 实时PCR; 实时荧光PCR检测方法; 探测效率; 有效管理; 种子检疫; real-time fluorescence polymerase chain reaction; bacterial grain rot; detection; rice; Burkholderia glumae
• ISSN: 1672-6308; 1876-4762
• DOI: 10.1016/S1672-6308(08)60073-6

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 1×10^4 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.