A Sequencing-Based Phylogenetic Analysis of Various Strains of Watermelon Silver Mottle Virus in Northern China and Their One-Step Detection Using Reverse Transcription Loop-Mediated Isothermal Amplification

Author:

Qiao Ning1ORCID,Jiang Miao1,Chen Yuxing1,Wang Xingcui1,Chi Wenjuan1,Li Shoujie1,Zhu Xiaoping2ORCID,Sun Xiaoan13

Affiliation:

1. Facility Horticulture Laboratory of Universities in Shandong, Weifang University of Science and Technology, Shouguang, Shandong, China

2. College of Plant Protection, Shandong Agricultural University, Taian, Shandong, China

3. Division of Plant Industry, Florida Department of Agriculture and Consumer Services, Gainesville, FL, U.S.A. (Retired)

Abstract

Watermelon silver mottle virus (WSMoV), a potentially invasive virus, is known to reduce the yield and degrade the quality of infected crops in Cucurbitaceae and Solanaceae families, resulting in significant economic losses in limited areas of several Asian countries. WSMoV, previously detected on various crops in southern China, has now become more prevalent on watermelon and sweet pepper in the northern cities of China for the first time. A sequencing-based phylogenetic analysis has confirmed that the viral strains infecting cucumber, watermelon, and sweet pepper plants in Shandong Province are most closely related to those isolated from Guangdong, Guangxi, and Taiwan, suggesting a farther and continuous spread of WSMoV throughout China. To develop a fast, accurate, and practical protocol for WSMoV detection, we designed a set of primers from the conserved sequence of the WSMoV nucleocapsid protein (N) gene for a one-step assay based on reverse transcription loop-mediated isothermal amplification (RT-LAMP). The RT-LAMP assay was performed successfully for 50 min at 61°C and exhibited a highly specific result without cross-reactions with other similar viruses and a sensitivity that is 100-fold higher than that of the traditional RT-PCR. The confirmation of 26 WSMoV suspect samples collected from various regions in Shandong through the RT-LAMP testing has demonstrated that the assay is suitable and practical for detection of WSMoV in both laboratory and field settings.

Funder

Weifang University of Science and Technology Doctoral Start-Up Fund

Shandong Major S&T Innovation Project

Shandong Provincial Agricultural Major Applied Technology Innovation Project

Shandong Province Innovation and Entrepreneurship Training Programme for College Students

Publisher

Scientific Societies

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