苹果茎沟病毒的RT-PCR检测及其在我国苹果产区的分布

为开发苹果茎沟病毒(Apple stem grooving virus,ASGV)田间样本RT-PCR检测方法以及揭示我国AS-GV的发生情况,本研究以染病的苹果组培苗为试材,对ASGV RT-PCR检测方法进行了优化并利用优化的检测体系对我国苹果产区ASGV的发生情况进行了检测。结果表明:引物ASGVS特异性最好,优化的RT-PCR检测体系能够在4,7和11月份检测果树树皮组织的带毒情况,灵敏度达到能够检测2.5×10-2μg新鲜样本,适于苹果ASGV田间样本检测。通过对我国苹果产区ASGV的检测发现,被检测的13个省(市/区)苹果上几乎都有ASGV发生,只有甘肃省泾川样本未检测到该病毒,采集的327个样本带毒率达73.7%。

RT-PCR detection and distribution of ASGV in the apple producing areas of China

In order to develop the RT-PCR method for test of Apple stem grooving virus(ASGV) in field and make clear the occurrence of ASGV in China,RT-PCR detection system of ASGV was modified using the apple tissue cultures infected by ASGV as material and the distribution of ASGV in China was detected by the modified method.The result showed that the primer ASGVS had the highest specificity and ASGV could be detected by the modified RT-PCR method using apple bark tissue in April,July and November.The sensitivity of the optimized RT-PCR method could reach as less as 2.5×10-2 μg fresh sample,which was applicable for field testing.The detection of ASGV in apple producing areas of China clarified that ASGV occurred widely in all of the 23 regions in 13 provinces(municipolity/autonomous region) of China except for Jingchuan of Gansu province.The positive rate of ASGV in 327 samples was 73.7%.