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Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (12): 2497-2505.doi: 10.16420/j.issn.0513-353x.2021-0147

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Real-time Fluorescence Quantitative PCR Detection of Plantago Asiatica Mosaic Virus in Lilies

SONG Meng, XU Leifeng(), CAO Yuwei, YANG Panpan, BI Mengmeng, HE Guoren, TANG Yuchao, WANG Jing, MING Jun()   

  1. Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China
  • Received:2021-04-12 Revised:2021-11-25 Published:2022-01-04
  • Contact: XU Leifeng,MING Jun E-mail:xuleifeng@caas.cn;mingjun@caas.cn

Abstract:

As a potentially harmful virus,plantago asiatica mosaic virus(PlAMV)has been reported to infect lilies and other crops in China,so it is urgent to establish a monitoring and control system to prevent its spread. In this study,specific primer pair CP-2F/R were designed based on the conserved region of coat protein(CP)gene of PlAMV. By optimizing concentration of primers and annealing temperature,an efficient real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)was established with optimal primer concentration of 10 μmol · L-1 and optimal annealing temperature of 59.4℃. The sensitivity of this assay was 100 times higher than that of conventional RT-PCR,and the limit of detection by RT-qPCR was 13 copies · μL-1. The established RT-qPCR was used to detect PlAMV in 75 lily samples,with conventional RT-PCR used as a reference method,and the results showed that more positive samples were detected by RT-qPCR (29,38.67%)than by RT-PCR(21,28.00%).

Key words: lily, plantago asiatica mosaic virus, real-time fluorescence quantitative PCR

CLC Number: