西瓜果腐病种子带菌的PCR检测研究

Detection of Acidovorax avenae subsp. citrulli in Watermelon Seed Using Ploymerase Chain Reaction

基于传统的PCR检测技术,研究了西瓜果腐病种子带菌的PCR检测方法。结果表明,磷酸缓冲液更适合于作为带菌种子的浸提液,PVPP可去除种子残留物,提高检测的灵敏度。使用本检测方法,摇培时间在2h以上时,其时间的长短不影响自然感染带菌种子的检测;能检测到的西瓜种子最低带菌率为1‰,即1000粒种子中存在1粒带菌的种子;而且能对自然感染的单粒种子进行检测,得到种子样品带菌率的数据。本研究所获得的检测方法简便实用,能有效应用于西瓜商品种子西瓜果腐病菌的检测。

Based on the classical PCR technique, the testing method for detection ofAcidovorax avenae subsp. citrulli in watermelon seed has been reported here. It was shown that PBS was more suitable to extract bacterial from watermelon seeds and PVPP could dispose of seed residue and improve the testing sensitivity. The time did not influence results of testing for watermelon seeds infected by nature when it is over 2 hours. The testing sensitivity can reach 1‰ of infected seeds,namely there was 1 infected seed in 1 000 seeds. 1 infected seed can be tested and the data of infected seeds percent can be got with this method. This demonstrate that the method in this study is simple and practicable and can be used to detection Aac in watermelon seeds.