Establishment of a Serology- and Molecular-Combined Detection System for Youcai Mosaic Virus and Its Application in Various Host Plants
Author:
Feng Chenwei1,
Hua Yanhong1,
Liu Duxuan1,
Chen Haoyu1,
Wu Mingjie1,
Hua Jing1,
Zhang Kun12ORCID
Affiliation:
1. Department of Plant Pathology, College of Plant Protection, Yangzhou University, Yangzhou 225009, China
2. Joint International Research Laboratory of Agriculture and Agri-Product Safety, Ministry of Education of China, Yangzhou University, Yangzhou 225009, China
Abstract
The youcai mosaic virus (YoMV) can infect a diverse array of crop species, such as Raphanus sativus, Brassica napus, Solanum nigrum, and Rehmannia glutinosa, causing substantial economic damage. This study aimed to develop a rapid, sensitive, and economical diagnostic method for YoMV. We successfully expressed and purified the recombinant His-CPYoMV-YZ protein in E. coli BL21, which was used to immunize New Zealand White rabbits, generating high-titer polyclonal antibodies (PAb-CPYoMV-YZ). Additionally, a serological-based reverse transcription loop-mediated isothermal amplification (S-RT-LAMP) assay was refined, combining serological and molecular detection techniques to enhance practicality. Utilizing PAb-CPYoMV-YZ, we developed four techniques for detecting YoMV: Western blot, dot immunoblotting assay, enzyme-linked immunosorbent assay (ELISA), and S-RT-LAMP. YoMV isolates from various regions and hosts were analyzed. The results indicated that PAb-CPYoMV-YZ was highly effective in detecting YoMV across a range of hosts and isolates from diverse regions. This study fills an important gap in the serological detection of YoMV and provides a practical tool for on-site diagnosis and control of YoMV infection.
Funder
National Natural Science Foundation of China
Excellent Youth Fund of Jiangsu Natural Science Foundation
Jiangsu Province Agricultural Science and technology independent innovation fund project
Postgraduate Research and Practice Innovation Program of the Jiangsu Province
Chinese Government Scholarship
Young and Middle-aged Academic Leaders of the “Qinglan Project” of Yangzhou University
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