{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T17:55:31Z","timestamp":1784224531520,"version":"3.55.0"},"reference-count":38,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,2,15]],"date-time":"2026-02-15T00:00:00Z","timestamp":1771113600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Science and Technology Council (NSTC), Taiwan","award":["MOST 106-3114-B-005-002"],"award-info":[{"award-number":["MOST 106-3114-B-005-002"]}]},{"name":"National Science and Technology Council (NSTC), Taiwan","award":["MOST 107-2321-B-005-004"],"award-info":[{"award-number":["MOST 107-2321-B-005-004"]}]},{"name":"National Science and Technology Council (NSTC), Taiwan","award":["MOST 108-2321-B-005-016"],"award-info":[{"award-number":["MOST 108-2321-B-005-016"]}]},{"name":"National Science and Technology Council (NSTC), Taiwan","award":["MOST 109-2321-B-005-028"],"award-info":[{"award-number":["MOST 109-2321-B-005-028"]}]},{"name":"Advanced Plant and Food Crop Biotechnology Center"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Climate warming has facilitated the expansion of insect vectors and plant viral pathogens, leading to increased incidence of viral diseases in crops. Cucurbit crops, including cucumber (Cucumis sativus), melon (Cucumis melo), squash (Cucurbita pepo), and watermelon (Citrullus lanatus), are of major economic importance worldwide, but their production is severely threatened by viral infections. Among the most damaging viruses are zucchini yellow mosaic virus (ZYMV; genus Potyvirus), transmitted by aphids, and melon yellow spot virus (MYSV; genus Orthotospovirus), transmitted by thrips, both of which cause significant yield losses in Asia, including Taiwan. Previously, an attenuated ZYMV mutant, ZAC, was shown to confer effective cross-protection against ZYMV in several cucurbit species. In the present study, we engineered a recombinant virus, ZAC-MYnp, by inserting the nucleocapsid protein (NP) open reading frame of MYSV into the ZAC genome. ZAC-MYnp retained the attenuated phenotype of ZAC and remained effective in protecting against ZYMV infection, with protection rates of 70.4% and 87.0% in zucchini and muskmelon plants, respectively. In addition, under both mechanical and thrips-mediated challenge conditions, ZAC-MYnp significantly reduced MYSV symptom severity in muskmelon, with a protection rate of 66.7% and a protective efficacy of 79.0%, respectively. These results demonstrate that ZAC-derived recombinant viruses can function as a bivalent viral vaccine, offering dual protection against an aphid-borne potyvirus and a thrips-borne orthotospovirus. Our study highlights the feasibility of using a bivalent recombinant vaccine to manage two distinct insect-borne viruses in cucurbit crops.<\/jats:p>","DOI":"10.3390\/v18020250","type":"journal-article","created":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T11:11:28Z","timestamp":1771240288000},"page":"250","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Generation of a Bivalent Recombinant Vaccine Conferring Dual Protection Against Potyvirus and Orthotospovirus in Cucurbits"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4422-5469","authenticated-orcid":false,"given":"Tsung-Chi","family":"Chen","sequence":"first","affiliation":[{"name":"Department of Medical Laboratory Science and Biotechnology, Asia University, Wufeng Dist., Taichung City 413305, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ya-Chi","family":"Kang","sequence":"additional","affiliation":[{"name":"Department of Medical Laboratory Science and Biotechnology, Asia University, Wufeng Dist., Taichung City 413305, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thi-Ngoc-Bich","family":"Tran","sequence":"additional","affiliation":[{"name":"Department of Plant Pathology, National Chung Hsing University, South Dist., Taichung City 402202, Taiwan"},{"name":"Faculty of Agronomy, Nong Lam University, Ho Chi Minh City 70000, Vietnam"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li-Hsin","family":"Huang","sequence":"additional","affiliation":[{"name":"Agricultural Chemicals Research Institute, Ministry of Agriculture, Wufeng Dist., Taichung City 413001, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chian-Chi","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Medical Laboratory Science and Biotechnology, Asia University, Wufeng Dist., Taichung City 413305, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5849-6753","authenticated-orcid":false,"given":"Shyi-Dong","family":"Yeh","sequence":"additional","affiliation":[{"name":"Department of Plant Pathology, National Chung Hsing University, South Dist., Taichung City 402202, Taiwan"},{"name":"Advanced Plant and Food Crop Biotechnology Center, National Chung Hsing University, South Dist., Taichung City 402202, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1094\/PDIS-11-09-0742","article-title":"Double-virus resistance of transgenic oriental melon conferred by untranslatable chimeric construct carrying partial coat protein genes of two viruses","volume":"94","author":"Wu","year":"2010","journal-title":"Plant Dis."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Peng, J.C., Chen, T.C., Raja, J.A.J., Yang, C.F., Chien, W.C., Lin, C.H., Liu, F.L., Wu, H.W., and Yeh, S.D. (2014). Broad-spectrum transgenic resistance against distinct tospovirus species at the genus level. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0096073"},{"key":"ref_3","first-page":"557","article-title":"Mosaic diseases in Canary Islands, West Africa, and Gribraltar","volume":"39","author":"McKinney","year":"1929","journal-title":"J. Agric. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1007\/BF01980475","article-title":"M II-16, an artificial symptomless mutant of tobacco mosaic virus for seedling inoculation of tomato crops","volume":"78","author":"Rast","year":"1971","journal-title":"Neth. J. Plant Pathol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Pechinger, K., Chooi, K.M., MacDiarmid, R.M., Harper, S.J., and Ziebell, H. (2019). A new era for mild strain cross-protection. Viruses, 11.","DOI":"10.3390\/v11070670"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1094\/PD-90-0067","article-title":"Effectiveness of an attenuated Zucchini yellow mosaic virus isolate for cross-protecting cucumber","volume":"90","author":"Kosaka","year":"2006","journal-title":"Plant Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1007\/s10327-011-0318-x","article-title":"Attenuated plant viruses: Preventing virus diseases and understanding the molecular mechanism","volume":"77","author":"Nishiguchi","year":"2011","journal-title":"J. Gen. Plant Pathol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1094\/PHYTO-97-3-0287","article-title":"Modifications of the HC-Pro of Zucchini yellow mosaic potyvirus for generation of attenuated mutants for cross protection against severe infection","volume":"97","author":"Lin","year":"2007","journal-title":"Phytopathology"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"944","DOI":"10.1094\/MPMI-04-14-0116-R","article-title":"Genetic analyses of the FRNK motif function of Turnip mosaic virus uncover multiple and potentially interactive pathways of cross-protection","volume":"27","author":"Kung","year":"2014","journal-title":"Mol. Plant Microbe Interact."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1094\/PHYTO-01-14-0018-R","article-title":"Generation of an attenuated, cross-protective Pepino mosaic virus variant through alignment-guided mutagenesis of the viral capsid protein","volume":"105","author":"Chewachong","year":"2015","journal-title":"Phytopathology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/s12985-019-1143-7","article-title":"Evaluation of Potato virus X mild mutants for cross protection against severe infection in China","volume":"16","author":"Cong","year":"2019","journal-title":"Virol. J."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1094\/PD-72-0375","article-title":"Control of Papaya ringspot virus by cross protection","volume":"72","author":"Yeh","year":"1988","journal-title":"Plant Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1094\/PD-75-0203","article-title":"Effectiveness of cross protection by a mild strain of zucchini yellow mosaic virus in cucumber, melon, and squash","volume":"75","author":"Wang","year":"1991","journal-title":"Plant Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1094\/PD-75-0208","article-title":"Control of zucchini yellow mosaic virus in squash by cross protection","volume":"75","author":"Lecoq","year":"1991","journal-title":"Plant Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1006\/viro.1998.9280","article-title":"Coat protein interactions involved in Tobacco mosaic tobamovirus cross-protection","volume":"248","author":"Lu","year":"1998","journal-title":"Virology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1094\/MPMI-23-1-0017","article-title":"Discriminating mutations of HC-Pro of zucchini yellow mosaic virus with differential effects on small RNA pathways involved in viral pathogenicity and symptom development","volume":"23","author":"Wu","year":"2010","journal-title":"Mol. Plant Microbe Interact."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1207","DOI":"10.1105\/tpc.11.7.1207","article-title":"Gene silencing without DNA: RNA-mediated cross-protection between viruses","volume":"11","author":"Ratcliff","year":"1999","journal-title":"Plant Cell"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1","DOI":"10.4172\/2329-9029.1000229","article-title":"Applications of virus-induced gene silencing (VIGS) in plant functional genomics studies","volume":"7","author":"Bekele","year":"2019","journal-title":"J. Plant Biochem. Physiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1016\/j.virol.2009.01.039","article-title":"Apple latent spherical virus vectors for reliable and effective virus-induced gene silencing among a broad range of plants including tobacco, tomato, Arabidopsis thaliana, cucurbits, and legumes","volume":"386","author":"Igarashi","year":"2009","journal-title":"Virology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/j.virusres.2013.06.015","article-title":"Development of apple latent spherical virus-based vaccines against three tospoviruses","volume":"176","author":"Taki","year":"2013","journal-title":"Virus Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1016\/j.virol.2013.08.019","article-title":"Preventive and curative effects of Apple latent spherical virus vectors harboring part of the target virus genome against potyvirus and cucumovirus infections","volume":"446","author":"Tamura","year":"2013","journal-title":"Virology"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1459","DOI":"10.1094\/PDIS-09-24-1949-RE","article-title":"A point mutation in pepper veinal mottle virus 6K1 protein yields a stable attenuated strain for engineering virus resistance in pepper plants","volume":"109","author":"Deng","year":"2025","journal-title":"Plant Dis."},{"key":"ref_23","unstructured":"Ministry of Agriculture, Executive Yuan (2026, January 29). Available online: https:\/\/agrstat.moa.gov.tw\/sdweb\/public\/inquiry\/InquireAdvance.aspx."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1007\/s10658-011-9746-x","article-title":"Emerging threat of thrips-borne Melon yellow spot virus on melon and watermelon in Taiwan","volume":"130","author":"Peng","year":"2011","journal-title":"Eur. J. Plant Pathol."},{"key":"ref_25","first-page":"259","article-title":"Molecular characterization and serological diagnosis of the new emerging Cucurbit chlorotic yellows virus in Taiwan","volume":"22","author":"Li","year":"2013","journal-title":"Plant Pathol. Bull."},{"key":"ref_26","first-page":"1","article-title":"Aphid transmissibility and cross-protection effectiveness of an attenuated mutant of Zucchini yellow mosaic virus","volume":"52","author":"Chao","year":"2010","journal-title":"Plant Prot. Bull."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Huang, C.H., Hsiao, W.R., Huang, C.W., Chen, K.C., Lin, S.S., Chen, T.C., Raja, J.A., Wu, H.W., and Yeh, S.D. (2015). Two novel motifs of Watermelon silver mottle virus NSs protein are responsible for RNA silencing suppression and pathogenicity. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0126161"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1094\/PHYTO-07-23-0227-R","article-title":"Leucine 127 of cucurbit chlorotic yellows virus P22 is crucial for its RNA silencing suppression activity and pathogenicity","volume":"114","author":"Kang","year":"2024","journal-title":"Phytopathology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1085","DOI":"10.1007\/s00705-010-0688-y","article-title":"Serological relationship between Melon yellow spot virus and Watermelon silver mottle virus and differential detection of the two viruses in cucurbits","volume":"155","author":"Chen","year":"2010","journal-title":"Arch. Virol."},{"key":"ref_30","first-page":"163","article-title":"Characterization of Zucchini yellow mosaic virus (ZYMV) isolates collected from Taiwan by host reactions, serology, and RT-PCR","volume":"40","author":"Lin","year":"1998","journal-title":"Plant Prot. Bull."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Wang, G.D., Lin, C.C., and Chen, T.C. (2024). Development of attenuated viruses for effective protection against pepper veinal mottle virus in tomato crops. Viruses, 16.","DOI":"10.3390\/v16050687"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/j.jviromet.2005.05.014","article-title":"Purification and serological analyses of tospoviral nucleocapsid proteins expressed by Zucchini yellow mosaic virus vector in squash","volume":"129","author":"Chen","year":"2005","journal-title":"J. Virol. Methods"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1046\/j.1461-9563.2002.00132.x","article-title":"A molecular identification key for economically important thrips species (Thysanoptera: Thripidae) using direct sequencing and a PCR-RFLP-based approach","volume":"4","author":"Brunner","year":"2002","journal-title":"Agric. For. Entomol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1111\/j.1365-3059.2011.02484.x","article-title":"Genomic characterization of Calla lily chlorotic spot virus and design of broad-spectrum primers for detection of tospoviruses","volume":"61","author":"Chen","year":"2012","journal-title":"Plant Pathol."},{"key":"ref_35","first-page":"1055","article-title":"Field evaluation of vaccine efficacy","volume":"63","author":"Orenstein","year":"1985","journal-title":"Bull. World Health Organ."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1296","DOI":"10.1094\/PHYTO-96-1296","article-title":"Identification of common epitopes on a conserved region of NSs proteins among tospoviruses of Watermelon silver mottle virus serogroup","volume":"96","author":"Chen","year":"2006","journal-title":"Phytopathology"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1583","DOI":"10.1094\/PHYTO-01-23-0019-R","article-title":"Concurrent control of two aphid-borne potyviruses in cucurbits by two-in-one vaccine","volume":"113","author":"Tran","year":"2023","journal-title":"Phytopathology"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Tran, T.N.B., Huang, C.H., Cheng, H.W., Liao, C.C., Raja, J.A.J., and Yeh, S.D. (2026). Development of an effective mild chimeric virus for the concurrent control of two important aphid-transmitted viruses in cucurbits. Phytopathology.","DOI":"10.1094\/PHYTO-07-25-0245-R"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/18\/2\/250\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T11:23:54Z","timestamp":1771241034000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/18\/2\/250"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,15]]},"references-count":38,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["v18020250"],"URL":"https:\/\/doi.org\/10.3390\/v18020250","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,15]]}}}