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Virus-induced gene silencing (VIGS) was employed to elucidate the function of AO in <jats:italic>N. benthamiana<\/jats:italic>. The tobacco rattle virus (TRV)-mediated VIGS resulted in an efficient silencing of the <jats:italic>NbAO<\/jats:italic> gene, i.e., 97.5% and 78.8% in relative quantification as compared to the control groups (TRV::eGFP- and the mock-inoculated plants), respectively. In addition, AO enzymatic activity decreased in the TRV::NtAO-silenced plants as compared to control. TRV::NtAO-mediated <jats:italic>NbAO<\/jats:italic> silencing induced a greater reduction in plant height by 15.2% upon CMV infection. CMV titer at 3 dpi was increased in the systemic leaves of <jats:italic>NbAO<\/jats:italic>-silenced plants (a 35-fold change difference as compared to the TRV::eGFP-treated group). Interestingly, CMV and TRV titers vary in different parts of systemically infected <jats:italic>N. benthamiana<\/jats:italic> leaves. In TRV::eGFP-treated plants, CMV accumulated only at the top half of the leaf, whereas the bottom half of the leaf was \u201coccupied\u201d by TRV. In contrast, in the <jats:italic>NbAO<\/jats:italic>-silenced plants, CMV accumulated in both the top and the bottom half of the leaf, suggesting that the silencing of the <jats:italic>NbAO<\/jats:italic> gene resulted in the expansion of the spreading area of CMV. Our data suggest that the <jats:italic>AO<\/jats:italic> gene might function as a resistant factor against CMV infection in <jats:italic>N. benthamiana<\/jats:italic>.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1007\/s00425-023-04313-x","type":"journal-article","created":{"date-parts":[[2024,1,16]],"date-time":"2024-01-16T13:02:08Z","timestamp":1705410128000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Silencing of a Nicotiana benthamiana ascorbate oxidase gene reveals its involvement in resistance against cucumber mosaic virus"],"prefix":"10.1007","volume":"259","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0800-6584","authenticated-orcid":false,"given":"Reshma","family":"Ahmed","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7762-5138","authenticated-orcid":false,"given":"Athanasios","family":"Kaldis","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0369-4904","authenticated-orcid":false,"given":"Andreas","family":"Voloudakis","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,1,16]]},"reference":[{"key":"4313_CR1","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1016\/S1369-5266(99)80022-3","volume":"2","author":"DC Baulcombe","year":"1999","unstructured":"Baulcombe DC (1999) Fast forward genetics based on virus-induced gene silencing. 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