基于短引物扩增应用的龙牙百合主要病毒鉴定

Identification of Main Viruses in Lilium brownii var.viridulum Based on Short Primer Amplification

为确定龙牙百合所感染病毒种类,有效减少百合病毒性病害带来的损失,采用随机取样法在湖南省隆回县取染病植株样本9株,以百合斑驳病毒(Lily mottle virus,LMoV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)和百合无症病毒(Lily symptomless virus,LSV)3种百合主要病毒已公布序列设计短引物,采用95℃预变性15 min,95℃变性10 s、60℃退火30 s、72℃延伸30 s,45个循环进行RT-qPCR。扩增结果为:LMoV-99、LMoV-101引物扩增产物的片段长度在100 bp左右,符合预期设计所能扩增出的片段长度,而LSV-169、CMV-70引物扩增产物的片段长度在50 bp以下,不符合预期片段长度,而LSV-70基本没有获得PCR产物。检测结果为:龙牙百合感染病毒为百合斑驳病毒,未感染黄瓜花叶病毒和百合无症病毒。

In order to determine the type of virus infected by Viruses of Lilium brownii var.viridulum,reduced effectively the loss caused by lily viral diseases.We used a random sampling method to sample 10 infected samples in Longhui County.We used the published sequences of three major Lily viruses which including Lily mottle virus,Cucumber mosaic virus,and Lily symptomless virus to design the short primers.RT-qPCR was performed under the conditions of 95℃ pre-denaturation for 15 min at 95℃ denaturation for 10 s at 60℃ annealing for 30 s,and 72℃ extension for 30 s.The results showed that the fragment length of LMoV-99 and LMoV-101 primers was about 100 bp,which was in line with the expected fragment length,and that of LSV-169 and CMV-70 primers was less than 50 bp,did not meet the expected fragment length,and LSV-70 did not obtain PCR products.The results of the test showed that Lily mottle virus was infected with iruses of Lilium brownii var.viridulum,and Cucumber mosaic virus and lily symptomless virus were not infected.