Journal of Integrative Agriculture ›› 2020, Vol. 19 ›› Issue (5): 1283-1291.DOI: 10.1016/S2095-3119(19)62755-3

所属专题: 植物病理合辑Plant Protection—Plant Pathology 植物病毒合辑Plant Virus

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  • 收稿日期:2019-02-17 出版日期:2020-04-01 发布日期:2020-03-25

Monoclonal antibody-based serological detection of potato virus M in potato plants and tubers

ZHANG Yu1, GAO Yan-ling2, HE Wan-qin1, WANG Ya-qin1, QIAN Ya-juan1, ZHOU Xue-ping1, 3, WU Jian-xiang1  
  

  1. 1 State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310058, P.R.China
    2 Potato Research Institute of Heilongjiang Academy of Agricultural Sciences, Harbin 150086, P.R.China
    3 State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China
  • Received:2019-02-17 Online:2020-04-01 Published:2020-03-25
  • Contact: Correspondence WU Jian-xiang, Tel: +86-571-88982250, E-mail: wujx@zju.edu.cn; ZHOU Xue-ping, E-mail: zzhou@zju.edu.
  • About author:ZHANG Yu, E-mail: 380175146@qq.com; cn
  • Supported by:
    We are grateful to Dr. Ding Xinshun (Samuel Roberts Noble Foundation [Retired], Ardmore, USA) for his valuable comments and manuscript edits. This work was supported by the National Key Research and Development Project of China (2017YFD0201604) and the National Natural Science Foundation of China (31571976).

Abstract:

Potato virus M (PVM) is one of the common and economically important potato viruses in potato-growing regions worldwide.  To investigate and control this viral disease, efficient and specific detection techniques are needed.  In this study, PVM virions were purified from infected potato plants and used as the immunogen to produce hybridomas secreting PVM-specific monoclonal antibodies (MAbs).  Four highly specific and sensitive murine MAbs, i.e., 1E1, 2A5, 8A1 and 17G8 were prepared through a conventional hybridoma technology.  Using these four MAbs, we have developed an antigen-coated plate (ACP)-ELISA, a dot-ELISA and a Tissue print-ELISA for detecting PVM infection in potato plants and tubers.  PVM could be detected in infected potato plant tissue crude extracts diluted at 1:10 240 (w/v, g mL–1) by the dot-ELISA or at 1:163 840 (w/v, g mL–1) by the ACP-ELISA.  The Tissue print-ELISA is the quickest and easiest assay among the three established serological assays and is more suitable for onsite large-scale sample detection.  Detection results of the field-collected samples showed that PVM is currently widespread in the Yunnan and the Heilongjiang provinces in China.  The field sample test results of the developed serological assays were supported by the results from RT-PCR and DNA sequencing.  We consider that the newly established ACP-ELISA, dot-ELISA and Tissue print-ELISA can benefit PVM detection in potato plant and tuber samples and field epidemiological studies of PVM.  These assays can also facilitate the production of virus-free seed potatoes and breeding for PVM-resistant potato cultivars, leading to the successful prevention of this potato viral disease.
 

Key words: potato virus M ,  monoclonal antibody ,  serological assay ,  antigen-coated plate (ACP)-ELISA ,  dot-ELISA ,  Tissue print-ELISA