瓜类细菌性果腐病病原IAS-PCR检测研究

Study on the detection of pathogen IAS-PCR of bacterial fruit blotch of cucurbits

本研究采用免疫磁珠吸附与PCR相结合的技术,对瓜类细菌性果腐病病原快速检测方法进行了研究。对新疆昌吉、海南、上海等地的西、甜瓜细菌性果腐病病叶、种子进行检测结果表明,用特异性引物,带菌病叶、种子均出现360bp特异条带,该方法对该菌悬浮液最低检出量为1×10cfu/mL,检出种子最低带菌率1‰。与常规PCR方法相比,检测时间周期为4h,且不需进行细菌DNA提取,灵敏度高出常规PCR100倍。这一方法的建立对于田间该病原的准确诊断,控制西、甜瓜种子带菌传播及进出口西、甜瓜种子检疫将起到重要作用。

Seeds and plant of gourd with bacterial fruit blotch(BFB)pathogen are tested and studied in the way of combination of immunological and PCR techniques.Watermelon and muskmelon specimen with pathogen from Changji Xinjiang ,Hainan and Shanghai etc.are detected. The result shows that 360 bp specificity belt shows by all specimen with BFB pathogen treated with primer of specificity,the minimum detecting volume of extraction is 1×10 cfu/mL,and the minimum carrier rate of seeds is 1‰.Its sensitivity is 100 times higher than that of conventional PCR,its detection period is only 4 hours.The establishment of this method will be significant to control spread of watermelon or muskmelon seeds with pathogen and quarantine accurately imported or exported seeds.