| Construction and Characterization of a Full-Length Infectious cDNA Clone of a Strain of Watermelon Mosaic Virus Isolated from Melon |
Hae-Ryun Kwak1 
, Su-bin Hong1, Jeong-Eun Kim2, Hee-Seong Byun1, Bong-Choon Lee1, Hong-Soo Choi3, Mikyeong Kim3
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1Crop Protection Division, National Institute of Agricultural Sciences, Rural Development Administration, Wanju 55365, Korea
2Seed Testing & Research Center, Korea Seed & Variety Service, Gimcheon 39660, Korea
3Department of Plant Medicine, Chungbuk National University, Cheongju 28644, Korea |
Correspondence:
Hae-Ryun Kwak, Tel: +82-63-238-3303, Fax: +82-63-238-3838,
Email: hrkwakhahn@korea.kr
Mikyeong Kim, Tel: +82-43-261-2509, Fax: +82-43-271-4414,
Email: mkim00@cbnu.ac.kr
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Received: 15 July 2024 • Revised: 14 October 2024 • Accepted: 15 October 2024 |
| Abstract |
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Watermelon mosaic virus (WMV), a member of the genus Potyvirus, causes serious economic losses in cucurbit crops. For molecular biological studies of viruses, it is necessary to construct an infectious clone that can facilitate gene functional analysis. In this study, we constructed an infectious cDNA clone of WMV genomic RNA by Gibson assembly and evaluated its virulence and symptoms on a variety of host plants. A WMV isolate (WMV-IS-me2) collected from melon in Iksan, Jeonbuk Province, Korea in 2015 caused mosaic symptoms on leaves. Four overlapping PCR fragments of the full-length genome of this isolate and the pJL89 binary vector with overhanging ends of 40 bp were amplified by reverse transcription PCR and joined in a single isothermal reaction. The complete nucleotide sequence of the infectious cDNA clone of WMV was determined and the recombinant vector was transformed into Agrobacterium tumefaciens GV3101. Following agro-infiltration, an infectious clone (pWMV-M, KACC95145P) systemically induced mosaic symptoms on watermelon (Citrullus lanatus), zucchini (Cucurbita pepo), melon (Cucumis melo), and Nicotiana benthamiana plants. This infectious clone may be useful for screening cucurbit varieties resistant to WMV, as well as for studying viral gene function. |
| Key Words:
Gibson assembly, infectious cDNA clone, Potyvirus, watermelon mosaic virus |
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