{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T18:40:28Z","timestamp":1790361628585,"version":"4.1.0"},"reference-count":68,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2022,8,21]],"date-time":"2022-08-21T00:00:00Z","timestamp":1661040000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100010198","name":"Ministerio de Asuntos Econ\u00f3micos y Transformaci\u00f3n Digital, Gobierno de Espa\u00f1a","doi-asserted-by":"publisher","award":["BIO2013\u201049053\u2010R"],"award-info":[{"award-number":["BIO2013\u201049053\u2010R"]}],"id":[{"id":"10.13039\/501100010198","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Ministerio de Ciencia e Innovaci\u00f3n","doi-asserted-by":"publisher","award":["PID2019\u2010109380RBI00\/AEI\/10.13039\/501100011033"],"award-info":[{"award-number":["PID2019\u2010109380RBI00\/AEI\/10.13039\/501100011033"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bsppjournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molecular Plant Pathology"],"published-print":{"date-parts":[[2022,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Technology based on artificial small RNAs, including artificial microRNAs (amiRNAs), exploits natural RNA silencing mechanisms to achieve silencing of endogenous genes or pathogens. This technology has been successfully employed to generate resistance against different eukaryotic viruses. However, information about viral RNA molecules effectively targeted by these small RNAs is rather conflicting, and factors contributing to the selection of virus mutants escaping the antiviral activity of virus\u2010specific small RNAs have not been studied in detail. In this work, we transformed\n                    <jats:italic>Nicotiana benthamiana<\/jats:italic>\n                    plants with amiRNA constructs designed against the potyvirus plum pox virus (PPV), a positive\u2010sense RNA virus, and obtained lines highly resistant to PPV infection and others showing partial resistance. These lines have allowed us to verify that amiRNA directed against genomic RNA is more efficient than amiRNA targeting its complementary strand. However, we also provide evidence that the negative\u2010sense RNA strand is cleaved by the amiRNA\u2010guided RNA silencing machinery. Our results show that the selection pressure posed by the amiRNA action on both viral RNA strands causes an evolutionary explosion that results in the emergence of a broad range of virus variants, which can further expand in the presence, and even in the absence, of antiviral challenges.\n                  <\/jats:p>","DOI":"10.1111\/mpp.13258","type":"journal-article","created":{"date-parts":[[2022,8,21]],"date-time":"2022-08-21T22:09:54Z","timestamp":1661119794000},"page":"1640-1657","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Targeting of genomic and negative\u2010sense strands of viral\n                    <scp>RNA<\/scp>\n                    contributes to antiviral resistance mediated by artificial\n                    <scp>miRNAs<\/scp>\n                    and promotes the emergence of complex viral populations"],"prefix":"10.1111","volume":"23","author":[{"given":"Frida","family":"Mesel","sequence":"first","affiliation":[{"name":"Department of Plant Molecular Genetics, Centro Nacional de Biotecnolog\u00eda (CNB\u2010CSIC) Campus Universidad Aut\u00f3noma de Madrid  Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4816-0949","authenticated-orcid":false,"given":"Mingmin","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Plant Molecular Genetics, Centro Nacional de Biotecnolog\u00eda (CNB\u2010CSIC) Campus Universidad Aut\u00f3noma de Madrid  Madrid Spain"},{"name":"College of Horticulture and Plant Protection Inner Mongolia Agricultural University  Hohhot China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beatriz","family":"Garc\u00eda","sequence":"additional","affiliation":[{"name":"Department of Plant Molecular Genetics, Centro Nacional de Biotecnolog\u00eda (CNB\u2010CSIC) Campus Universidad Aut\u00f3noma de Madrid  Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4251-0821","authenticated-orcid":false,"given":"Carmen","family":"Sim\u00f3n\u2010Mateo","sequence":"additional","affiliation":[{"name":"Department of Plant Molecular Genetics, Centro Nacional de Biotecnolog\u00eda (CNB\u2010CSIC) Campus Universidad Aut\u00f3noma de Madrid  Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4861-9609","authenticated-orcid":false,"given":"Juan\u00a0Antonio","family":"Garc\u00eda","sequence":"additional","affiliation":[{"name":"Department of Plant Molecular Genetics, Centro Nacional de Biotecnolog\u00eda (CNB\u2010CSIC) Campus Universidad Aut\u00f3noma de Madrid  Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2022,8,21]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.279.5359.2113"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1105\/tpc.105.040725"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth0805-629"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-arplant-050312-120043"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2005.04.012"},{"key":"e_1_2_8_7_1","first-page":"e1130","article-title":"Fast\u2010forward generation of effective artificial small RNAs for enhanced antiviral defense in plants","volume":"3","author":"Carbonell A.","year":"2016","journal-title":"RNA & Disease"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/tpj.14466"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1094\/MPMI-05-18-0117-TA"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.3390\/plants9060669"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.103.027979"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature02697"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.111.172627"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2004.01.021"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.18.01224"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0955-0674(95)80096-4"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01377-08"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2011.00647.x"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btv534"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2021.08.027"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1006\/viro.2000.0762"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.phyto.45.062806.094434"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1106-1358"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10123-003-0142-z"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0011387"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.72.5.4072-4079.1998"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.286.5441.950"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009757"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.227.4691.1229"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1111\/mpp.12291"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1364-3703.2011.00747.x"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.05326-11"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00914-13"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.84.21.7542"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/0168-1702(89)90013-0"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1186\/1753-6561-8-S4-O24"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt068"},{"key":"e_1_2_8_38_1","volume-title":"Expresi\u00f3n de prote\u00ednas heter\u00f3logas en plantas por medio del virus de la sharka (PPV)","author":"Lucini C.","year":"2004"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2018100941"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7600340"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.112.207068"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/mss135"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature02873"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1255"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02383-06"},{"key":"e_1_2_8_46_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.abg2264"},{"key":"e_1_2_8_47_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2017.05.006"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02457-06"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.276.5318.1558"},{"key":"e_1_2_8_50_1","doi-asserted-by":"publisher","DOI":"10.1016\/bs.aivir.2014.11.006"},{"key":"e_1_2_8_51_1","doi-asserted-by":"publisher","DOI":"10.1094\/MPMI-07-19-0189-FI"},{"key":"e_1_2_8_52_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2006.10.005"},{"key":"e_1_2_8_53_1","doi-asserted-by":"publisher","DOI":"10.1105\/tpc.105.039834"},{"key":"e_1_2_8_54_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11240-013-0339-6"},{"key":"e_1_2_8_55_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tree.2008.06.005"},{"key":"e_1_2_8_56_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.80.5.2429-2436.2006"},{"key":"e_1_2_8_57_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbagrm.2011.05.011"},{"key":"e_1_2_8_58_1","doi-asserted-by":"publisher","DOI":"10.1038\/35030305"},{"key":"e_1_2_8_59_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-9861(69)90285-9"},{"key":"e_1_2_8_60_1","doi-asserted-by":"publisher","DOI":"10.1105\/tpc.010366"},{"key":"e_1_2_8_61_1","doi-asserted-by":"publisher","DOI":"10.1093\/pcp\/pcn043"},{"key":"e_1_2_8_62_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-9525(01)02367-8"},{"key":"e_1_2_8_63_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gki220"},{"key":"e_1_2_8_64_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.79.11.7050-7058.2005"},{"key":"e_1_2_8_65_1","doi-asserted-by":"publisher","DOI":"10.1111\/pbi.13278"},{"key":"e_1_2_8_66_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-phyto-020620-114550"},{"key":"e_1_2_8_67_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(85)90120-9"},{"key":"e_1_2_8_68_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2011.04.010"},{"key":"e_1_2_8_69_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002122"}],"container-title":["Molecular Plant Pathology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/mpp.13258","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/mpp.13258","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/bsppjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/mpp.13258","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,4]],"date-time":"2025-11-04T13:58:47Z","timestamp":1762264727000},"score":1,"resource":{"primary":{"URL":"https:\/\/bsppjournals.onlinelibrary.wiley.com\/doi\/10.1111\/mpp.13258"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,8,21]]},"references-count":68,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2022,11]]}},"alternative-id":["10.1111\/mpp.13258"],"URL":"https:\/\/doi.org\/10.1111\/mpp.13258","archive":["Portico"],"relation":{},"ISSN":["1464-6722","1364-3703"],"issn-type":[{"value":"1464-6722","type":"print"},{"value":"1364-3703","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,8,21]]},"assertion":[{"value":"2022-03-25","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-07-16","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-08-21","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}