园艺学报 ›› 2021, Vol. 48 ›› Issue (12): 2497-2505.doi: 10.16420/j.issn.0513-353x.2021-0147

• 新方法 • 上一篇    下一篇


宋蒙, 徐雷锋(), 曹雨薇, 杨盼盼, 毕蒙蒙, 何国仁, 唐玉超, 王静, 明军()   

  1. 中国农业科学院蔬菜花卉研究所,北京100081
  • 收稿日期:2021-04-12 修回日期:2021-11-25 发布日期:2022-01-04
  • 通讯作者: 徐雷锋,明军;
  • 基金资助:

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;


车前草花叶病毒(Plantago asiatica mosaic virus,PlAMV)病是侵染百合等作物的危险性病害,亟待建立其监测防控体系,以防止其扩散传播。根据PlAMV的外壳蛋白(Coat protein,CP)基因的保守序列设计特异性检测引物,通过对引物浓度和反应退火温度进行优化,建立了以CP-2 F/R为特异引物,引物浓度为10 μmol · L-1,退火温度为59.4 ℃的高效实时荧光定量PCR检测技术。该检测技术特异性强,灵敏度是RT-PCR的100倍,病毒最低检出浓度为13 copies · μL-1。对75份田间百合样品进行检测,结果显示:RT-qPCR方法检测(29份,38.67%)高于RT-PCR检测出的阳性样品(21份,28.00%)。

关键词: 百合, 车前草花叶病毒, 实时荧光定量PCR


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