蚜虫中携带马铃薯卷叶病毒检测方法的改进

Improved method detection Potato leafroll virus in aphids

关键词:  马铃薯卷叶病毒, 蚜虫, CP基因, 检测

Key words:  Potato leafroll virus, aphid, CP gene, detection

摘要:

马铃薯卷叶病毒(Potato leafroll virus,PLRV)对马铃薯生产的危害极大,是一种极为重要的马铃薯病毒病。RT-PCR是马铃薯卷叶病毒检测较为常用的方法,该方法检测准确率高、成本低、适用范围广。但在实际生产中其检测对象多为染病植株,对PLRV传播的主要介体桃蚜(Myzus persicae)的检测,则由于蚜虫体积小、RNA提取难度大、成本高、且不能复检,因而在生产中不能被广泛使用。该研究以马铃薯感病植株和带毒蚜虫为材料,利用改进的RNA提取方法从它们中提取到PLRV的RNA,并以CP基因设计特异性引物,进行PCR检测。结果表明:该方法提取的RNA完整性好,可用于蚜虫中PLRV检测,且同样适用于对马铃薯感病植株的检测。另外,通过对田间有翅蚜和无翅蚜携带PLRV情况进行检测发现,无翅蚜PLRV检出率为100%,有翅蚜PLRV检出率也高达60%,证明该体系在生产中的实用性。该研究使用改进的RNA提取方法,提取蚜虫中RNA,并利用RT-PCR进行了PLRV检测,与以前的方法相比简单实用,可被应用于生产检测中。该研究结果为马铃薯生产中PLRV的防控提供了一种新的手段。

Abstract:

In the potato production, Potato leafroll virus(PLRV)can be caused enormous harm,so that the PLRV is a very important disease in potato production. RT-PCR is a commonly used method for detection of the PLRV, and the detection method is high accuracy, low cost and widely used for detection of the PLRV in potato production. But in the actual production, the samples of detection often are the infected plants, and the Myzus persicae that is the main mediator of PLRV is not detected, because of the aphid's small volume, RNA extraction of high difficulty, high cost, and that can not be rechecked extraction. For these reasons, the detection method of RT-PCR can not often be used in the potato production. In this paper, we used potato susceptible plants and viruliferous aphids as experimental materials, and used the improved method of RNA extraction from which they were extracted by PLRV RNA. Then, we made detection with PCR, and the specific primers were designed that refer of CP gene. The experimental results showed that the RNA extraction of potato susceptible plants and viruliferous aphids was integral that could be used to detect the PLRV in aphids, and it also can be used to detect the PLRV in potato susceptible plants. In addition, we used this method to detect the field of aphids and wingless aphids carrying PLRV. Wingless aphids PLRV positive rate was 100%, the positive rate of PLRV of aphids was as high as 60%, which proved that there was a good pragmatism of the practicability of the system in potato production. In this study, we used the improved efficacious method to extract RNA, and used RT-PCR to detect the PLRV in aphids. Compared with the detection of method, the new method is simple and practical, it can be used in potato production test, and it is a new means for the prevention and control of the PLRV in the potato production.