{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T11:58:33Z","timestamp":1792670313600,"version":"4.5.0"},"reference-count":45,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2020,3,4]],"date-time":"2020-03-04T00:00:00Z","timestamp":1583280000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31572471"],"award-info":[{"award-number":["31572471"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Honey bee viruses are associated with honey bee colony decline. Israeli acute paralysis virus (IAPV) is considered to have a strong impact on honey bee survival. Phylogenetic analysis of the viral genomes from several regions of the world showed that various IAPV lineages had substantial differences in virulence. Chronic bee paralysis virus (CBPV), another important honey bee virus, can induce two significantly different symptoms. However, the infection characteristics and pathogenesis of IAPV and CBPV have not been completely elucidated. Here, we constructed infectious clones of IAPV and CBPV using a universal vector to provide a basis for studying their replication and pathogenesis. Infectious IAPV and CBPV were rescued from molecular clones of IAPV and CBPV genomes, respectively, that induced typical paralysis symptoms. The replication levels and expression proteins of IAPV and CBPV in progeny virus production were confirmed by qPCR and Western blot. Our results will allow further dissection of the role of each gene in the context of viral infection while helping to study viral pathogenesis and develop antiviral drugs using reverse genetics systems.<\/jats:p>","DOI":"10.3390\/ijms21051742","type":"journal-article","created":{"date-parts":[[2020,3,4]],"date-time":"2020-03-04T10:46:08Z","timestamp":1583318768000},"page":"1742","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["A Reverse Genetics System for the Israeli Acute Paralysis Virus and Chronic Bee Paralysis Virus"],"prefix":"10.3390","volume":"21","author":[{"given":"Sa","family":"Yang","sequence":"first","affiliation":[{"name":"Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China"},{"name":"Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture and Rural Affairs, Beijing 100093, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongxia","family":"Zhao","sequence":"additional","affiliation":[{"name":"Guangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Institute of Applied Biological Resources, Guangzhou 510260, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanchun","family":"Deng","sequence":"additional","affiliation":[{"name":"Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China"},{"name":"Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture and Rural Affairs, Beijing 100093, China"},{"name":"Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100093, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuai","family":"Deng","sequence":"additional","affiliation":[{"name":"Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China"},{"name":"Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture and Rural Affairs, Beijing 100093, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinling","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China"},{"name":"Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture and Rural Affairs, Beijing 100093, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingyun","family":"Diao","sequence":"additional","affiliation":[{"name":"Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China"},{"name":"Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture and Rural Affairs, Beijing 100093, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunsheng","family":"Hou","sequence":"additional","affiliation":[{"name":"Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China"},{"name":"Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture and Rural Affairs, Beijing 100093, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,3,4]]},"reference":[{"key":"ref_1","first-page":"303","article-title":"Importance of pollinators in changing landscapes for world crops","volume":"1608","author":"Klein","year":"2007","journal-title":"Proc. Biol. Sci."},{"key":"ref_2","first-page":"13","article-title":"Chinese agriculture\u2019s dependence on bee pollination: Plantation status of crops depending on bee pollination","volume":"8","author":"Sun","year":"2016","journal-title":"Chin. Agri. Sci. Bull."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Cornman, R.S., Tarpy, D.R., Chen, Y.P., Jeffreys, L., Lopez, D., Pettis, J.S., van Engelsdorp, D., and Evans, J.D. (2012). Pathogen webs in collapsing honey bee colonies. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0043562"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1255957","DOI":"10.1126\/science.1255957","article-title":"Bee declines driven by combined stress from parasites, pesticides, and lack of flowers","volume":"6229","author":"Goulson","year":"2015","journal-title":"Science"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1038\/nature20167","article-title":"Redefining the invertebrate RNA virosphere","volume":"540","author":"Shi","year":"2016","journal-title":"Nature"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1126\/science.1146498","article-title":"A metagenomic survey of microbes in honey bee colony collapse disorder","volume":"318","author":"Conlan","year":"2007","journal-title":"Science"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3428","DOI":"10.1099\/vir.0.83284-0","article-title":"Isolation and characterization of Israeli acute paralysis virus, a dicistrovirus affecting honeybees in Israel: Evidence for diversity due to intra- and inter-species recombination","volume":"88","author":"Maori","year":"2007","journal-title":"J. Gen. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1304","DOI":"10.1126\/science.1220941","article-title":"Global honey bee viral landscape altered by a parasitic mite","volume":"336","author":"Martin","year":"2012","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/j.scitotenv.2018.02.258","article-title":"Enhancement of chronic bee paralysis virus levels in honeybees acute exposed to imidacloprid: A Chinese case study","volume":"630","author":"Diao","year":"2018","journal-title":"Sci. Total Environ."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Singh, R., Abby, L.L., Edwin, G.R., Edward, C.H., Ostiguy, N., Vanengelsdorp, D., Lipkin, W.I., Depamphilis, C.W., Toth, A.L., and Cox-Foster, D.L. (2010). RNA viruses in hymenopteran pollinators: Evidence of inter-Taxa virus transmission via pollen and potential impact on non-Apis hymenopteran species. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0014357"},{"key":"ref_11","first-page":"151","article-title":"Varroa destructor is an effective vector of Israeli acute paralysis virus in the honeybee, Apis mellifera","volume":"1","author":"Pennacchio","year":"2011","journal-title":"J. Gen. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Francis, R.M., Nielsen, S.L., and Kryger, P. (2013). Varroa-virus interaction in collapsing honey bee colonies. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0057540"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/S0065-3527(07)70002-7","article-title":"Honey bee viruses","volume":"1","author":"Chen","year":"2007","journal-title":"Adv. Virus Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"S30","DOI":"10.1016\/j.jip.2009.06.014","article-title":"The acute bee paralysis virus\u2013Kashmir bee virus\u2013Israeli acute paralysis virus complex","volume":"103","author":"Cordoni","year":"2010","journal-title":"J. Invertebr. Pathol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2012","DOI":"10.3390\/v6052012","article-title":"Dynamics of the presence of Israeli acute paralysis virus in honey bee colonies with colony collapse disorder","volume":"6","author":"Hou","year":"2014","journal-title":"Viruses"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"6209","DOI":"10.1128\/JVI.00251-08","article-title":"Genetic analysis of Israel acute paralysis virus: Distinct clusters are circulating in the United States","volume":"13","author":"Palacios","year":"2008","journal-title":"J. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1016\/j.virol.2006.11.038","article-title":"Reciprocal sequence exchange between non-retro viruses and hosts leading to the appearance of new host phenotypes","volume":"2","author":"Maori","year":"2007","journal-title":"Virology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"8150","DOI":"10.1128\/JVI.00854-16","article-title":"Virion structure of Israeli acute bee paralysis virus","volume":"18","author":"Mullapudi","year":"2016","journal-title":"J. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"423493","DOI":"10.1155\/2015\/423493","article-title":"RNA 1 and RNA 2 genomic segments of chronic bee paralysis virus are infectious and induce chronic bee paralysis disease","volume":"2015","author":"Youssef","year":"2015","journal-title":"J. Immunol. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.virusres.2007.10.014","article-title":"Molecular characterization and phylogenetic analysis of chronic bee paralysis virus, a honey bee virus","volume":"132","author":"Olivier","year":"2008","journal-title":"Virus Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"S73","DOI":"10.1016\/j.jip.2009.06.017","article-title":"Nosema ceranae in European honey bees (Apis mellifera)","volume":"103","author":"Fries","year":"2010","journal-title":"J. Invertebr. Pathol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.pestbp.2017.10.009","article-title":"Metabolisation of thiamethoxam (a neonicotinoid pesticide) and interaction with the Chronic bee paralysis virus in honeybees","volume":"144","author":"Coulon","year":"2018","journal-title":"Pestic. Biochem. Physiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2282","DOI":"10.3390\/v5092282","article-title":"Chronic bee paralysis virus and Nosema ceranae experimental co-infection of winter honey bee workers (Apis mellifera L.)","volume":"9","author":"Toplak","year":"2013","journal-title":"Viruses"},{"key":"ref_24","first-page":"800","article-title":"In vitro culture of cells of the bee","volume":"11","author":"Giauffret","year":"1967","journal-title":"C. R. Acad. Sci. Hebd. Seances Acad. Sci. D"},{"key":"ref_25","first-page":"17","article-title":"Long-term maintenance of in vitro cultured honeybee (Apis mellifera) embryonic cells","volume":"1","author":"Monica","year":"2006","journal-title":"BMC Dev. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1111\/j.1365-2583.2010.01022.x","article-title":"Proteome profile and lentiviral transduction of cultured honey bee (Apis mellifera L.) cells","volume":"5","author":"Chan","year":"2010","journal-title":"Insect Mol. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1016\/0040-8166(94)90007-8","article-title":"Morphological analysis of honeybee antennal cells growing in primary cultures","volume":"4","author":"Gascuel","year":"1994","journal-title":"Tissue Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1007\/BF01187116","article-title":"Dissociated neurons of the pupal honeybee brain in cell culture","volume":"8","author":"Kreissl","year":"1992","journal-title":"J. Neurocytol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Hunter, W., Ellis, J., Vanengelsdorp, D., Hayes, J., Westervelt, D., Glick, E., Williams, M., Sela, I., Maori, E., and Pettis, J. (2010). Large-scale field application of RNAi technology reducing Israeli acute paralysis virus disease in honey bees (Apis mellifera, Hymenoptera: Apidae). PLoS Pathog., 12.","DOI":"10.1371\/journal.ppat.1001160"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3435","DOI":"10.1007\/s00705-014-2183-3","article-title":"Replication of Chinese sacbrood virus in primary cell cultures of Asian honeybee (Apis cerana)","volume":"12","author":"Xia","year":"2014","journal-title":"Arch. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Goblirsch, M.J., Spivak, M.S., and Kurtti, T.J. (2013). A cell line resource derived from honey bee (Apis mellifera) embryonic tissues. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0069831"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"22265","DOI":"10.1038\/srep22265","article-title":"In vivo and in vitro infection dynamics of honey bee viruses","volume":"6","author":"Dolezal","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3139","DOI":"10.1099\/0022-1317-83-12-3139","article-title":"Development of infectious transcripts and genome manipulation of black queen-cell virus of honey bees","volume":"12","author":"Benjeddou","year":"2002","journal-title":"J. Gen. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Lamp, B., Url, A., Seitz, K., Eichhorn, J., Riedel, C., Sinn, L.J., Indik, S., Ko\u00e8, H., and R\u00fcmenapf, T. (2016). Construction and rescue of a molecular clone of deformed wing virus (DWV). PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0164639"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"16274","DOI":"10.1038\/s41598-019-52822-1","article-title":"A molecular clone of chronic bee paralysis virus (CBPV) causes mortality in honey bee pupae (Apis mellifera)","volume":"1","author":"Seitz","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1016\/j.jip.2011.10.006","article-title":"Occurrence and prevalence of seven bee viruses in Apis mellifera and Apis cerana apiaries in China","volume":"1","author":"Ai","year":"2012","journal-title":"J. Invertebr. Pathol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1111\/j.1365-2583.2009.00847.x","article-title":"IAPV, a bee-affecting virus associated with Colony Collapse Disorder can be silenced by dsRNA ingestion","volume":"1","author":"Maori","year":"2009","journal-title":"Insect Mol. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Boncristiani, H.F., Evans, J.D., Chen, Y.P., Pettis, J., Murphy, C., Lopez, D.L., Simone-Finstrom, M., Strand, M., Tarpy, D.R., and Rueppell, O. (2013). In vitro infection of pupae with Israeli acute paralysis virus suggests disturbance of transcriptional homeostasis in honey bees (Apis mellifera). PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0073429"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1007\/s11626-009-9246-x","article-title":"Medium for development of bee cell cultures (Apis mellifera: Hymenoptera: Apidae)","volume":"2","author":"Hunter","year":"2010","journal-title":"In Vitro Cell. Dev. Biol. Anim."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.virusres.2008.06.006","article-title":"Functional analysis of structural motifs in dicistroviruses","volume":"139","author":"Nakashima","year":"2009","journal-title":"Virus Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1002\/wrna.52","article-title":"Cap and cap-binding proteins in the control of gene expression","volume":"2","author":"Topisirovic","year":"2011","journal-title":"Wiley Interdiscip. Rev. RNA"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"S120","DOI":"10.1016\/j.jip.2009.06.013","article-title":"Chronic bee paralysis: A disease and a virus like no other?","volume":"103","author":"Magali","year":"2010","journal-title":"J. Invertebr. Pathol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/j.virusres.2012.04.012","article-title":"Experimental infection of the honeybee (Apis mellifera L.) with the chronic bee paralysis virus (CBPV): Infectivity of naked CBPV RNAs","volume":"2","author":"Chevin","year":"2012","journal-title":"Virus Res."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Yang, S.A., Gayral, P., Zhao, H.X., Wu, Y.J., Jiang, X.J., Wu, Y.Y., Bigot, D., Wang, X.L., Yang, D.H., and Herniou, E.A. (2020). Occurrence and molecular phylogeny of honey bee viruses in Vespids. Viruses, 12.","DOI":"10.3390\/v12010006"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1139\/cjz-2014-0181","article-title":"Response of the honey bee (Apis mellifera) proteome to Israeli acute paralysis virus","volume":"9","author":"Michaud","year":"2015","journal-title":"Can. J. Zool."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/5\/1742\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:03:53Z","timestamp":1760173433000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/5\/1742"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,4]]},"references-count":45,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2020,3]]}},"alternative-id":["ijms21051742"],"URL":"https:\/\/doi.org\/10.3390\/ijms21051742","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,3,4]]}}}