{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T16:22:11Z","timestamp":1792167731143,"version":"4.4.0"},"reference-count":55,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2024,12,18]],"date-time":"2024-12-18T00:00:00Z","timestamp":1734480000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the Italian Ministry of Education, University and Research under Programme PON 2014\u20132020 and FSC"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Viola ucriana Erben &amp; Raimondo is a rare and endangered taxon, endemic to a limited area on Mount Pizzuta in northwestern Sicily, Italy. Its population is significantly threatened by anthropogenic activities, including fires, overgrazing, and habitat alterations. Temporary immersion systems (TISs) have proven effective for large-scale propagation in various protected species, offering potential for ex situ conservation and population reinforcement of V. ucriana. This study aimed to establish a bioreactor-based micropropagation protocol for shoot multiplication and compare the efficacy of a TIS with that of conventional solid culture medium (SCM). Three different plant growth regulators (PGRs) were also compared: 6-benzylaminopurine (BA), zeatin, and meta-topolin-9-riboside (mTR). The starting material originated from seeds collected from mother plants in their natural environment. The best growth outcomes (in terms of shoot multiplication, shoot length, and relative growth rate) were achieved using THE RITA\u00ae TIS, with BA (0.2 mg\/L) and mTR (0.5 or 0.8 mg\/L) outperforming SCM. Anomalous or hyperhydric shoots were observed with all zeatin treatments (especially with 0.8 mg\/L) in both the TIS and SCM, suggesting that this cytokinin is unsuitable for V. ucriana biomass production. The rooting phase was significantly improved by transferring propagules onto rockwool cubes fertilized with Hoagland solution. This approach yielded more robust roots in terms of number and length compared to the conventional agar-based medium supplemented with indole-3-butyric acid (IBA). Flow cytometry analysis confirmed the genetic fidelity of the regenerants from the optimal PGR treatments, showing that all plantlets maintained the diploid ploidy level of their maternal plants. Over 90% of the in vitro derived plantlets were successfully acclimatized to greenhouse conditions. This paper represents the first report of V. ucriana biomass multiplication using a RITA\u00ae bioreactor. The stability of the regenerants, confirmed by nuclei quantification via cytofluorimetry, provides guidance in establishing a true-to-type ex situ population, supporting conservation and future reinforcement efforts.<\/jats:p>","DOI":"10.3390\/plants13243530","type":"journal-article","created":{"date-parts":[[2024,12,18]],"date-time":"2024-12-18T08:56:32Z","timestamp":1734512192000},"page":"3530","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["RITA\u00ae Temporary Immersion System (TIS) for Biomass Growth Improvement and Ex Situ Conservation of Viola ucriana Erben &amp; Raimondo"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2521-928X","authenticated-orcid":false,"given":"Piergiorgio","family":"Capaci","sequence":"first","affiliation":[{"name":"Department of Biological and Environmental Sciences and Technologies (Di.S.Te.B.A.), University of Salento, Campus Ecotekne, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1409-2324","authenticated-orcid":false,"given":"Fabrizio","family":"Barozzi","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Sciences and Technologies (Di.S.Te.B.A.), University of Salento, Campus Ecotekne, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefania","family":"Forciniti","sequence":"additional","affiliation":[{"name":"Institute of Nanotechnology\u2014NANOTEC, Consiglio Nazionale delle Ricerche (CNR), Campus Ecotekne, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4218-2706","authenticated-orcid":false,"given":"Chiara","family":"Anglana","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Sciences and Technologies (Di.S.Te.B.A.), University of Salento, Campus Ecotekne, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9188-0511","authenticated-orcid":false,"given":"Helena","family":"Iuele","sequence":"additional","affiliation":[{"name":"Institute of Nanotechnology\u2014NANOTEC, Consiglio Nazionale delle Ricerche (CNR), Campus Ecotekne, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7432-9971","authenticated-orcid":false,"given":"Rita Annunziata","family":"Accogli","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Sciences and Technologies (Di.S.Te.B.A.), University of Salento, Campus Ecotekne, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5627-0120","authenticated-orcid":false,"given":"Angela","family":"Carra","sequence":"additional","affiliation":[{"name":"Institute of Biosciences and Bioresources, National Research Council (CNR-IBBR), Via Ugo La Malfa 153, 90146 Palermo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6493-9549","authenticated-orcid":false,"given":"Marcello Salvatore","family":"Lenucci","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Sciences and Technologies (Di.S.Te.B.A.), University of Salento, Campus Ecotekne, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9733-7088","authenticated-orcid":false,"given":"Loretta L.","family":"del Mercato","sequence":"additional","affiliation":[{"name":"Institute of Nanotechnology\u2014NANOTEC, Consiglio Nazionale delle Ricerche (CNR), Campus Ecotekne, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5388-0695","authenticated-orcid":false,"given":"Gian Pietro","family":"Di Sansebastiano","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Sciences and Technologies (Di.S.Te.B.A.), University of Salento, Campus Ecotekne, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1080\/00837792.2012.10670908","article-title":"Viola Section Melanium (Violaceae) in Italy. 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