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RITA® Temporary Immersion System (TIS) for Biomass Growth Improvement and Ex Situ Conservation of Viola ucriana Erben & Raimondo

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Title: RITA® Temporary Immersion System (TIS) for Biomass Growth Improvement and Ex Situ Conservation of Viola ucriana Erben & Raimondo
Authors: Piergiorgio Capaci, Fabrizio Barozzi, Stefania Forciniti, Chiara Anglana, Helena Iuele, Rita Annunziata Accogli, Angela Carra, Marcello Salvatore Lenucci, Loretta L. del Mercato, Gian Pietro Di Sansebastiano
Source: Plants, Vol 13, Iss 24, p 3530 (2024)
Publisher Information: MDPI AG, 2024.
Publication Year: 2024
Collection: LCC:Botany
Subject Terms: Viola ucriana, ex situ conservation, Temporary Immersion System (TIS), flow cytometry, Botany, QK1-989
More Details: Viola ucriana Erben & 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® 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® 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.
Document Type: article
File Description: electronic resource
Language: English
ISSN: 2223-7747
Relation: https://www.mdpi.com/2223-7747/13/24/3530; https://doaj.org/toc/2223-7747
DOI: 10.3390/plants13243530
Access URL: https://doaj.org/article/8dfac635fe7941c28915761160f3fac7
Accession Number: edsdoj.8dfac635fe7941c28915761160f3fac7
Database: Directory of Open Access Journals
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  Data: RITA® Temporary Immersion System (TIS) for Biomass Growth Improvement and Ex Situ Conservation of Viola ucriana Erben & Raimondo
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  Data: <searchLink fieldCode="AR" term="%22Piergiorgio+Capaci%22">Piergiorgio Capaci</searchLink><br /><searchLink fieldCode="AR" term="%22Fabrizio+Barozzi%22">Fabrizio Barozzi</searchLink><br /><searchLink fieldCode="AR" term="%22Stefania+Forciniti%22">Stefania Forciniti</searchLink><br /><searchLink fieldCode="AR" term="%22Chiara+Anglana%22">Chiara Anglana</searchLink><br /><searchLink fieldCode="AR" term="%22Helena+Iuele%22">Helena Iuele</searchLink><br /><searchLink fieldCode="AR" term="%22Rita+Annunziata+Accogli%22">Rita Annunziata Accogli</searchLink><br /><searchLink fieldCode="AR" term="%22Angela+Carra%22">Angela Carra</searchLink><br /><searchLink fieldCode="AR" term="%22Marcello+Salvatore+Lenucci%22">Marcello Salvatore Lenucci</searchLink><br /><searchLink fieldCode="AR" term="%22Loretta+L%2E+del+Mercato%22">Loretta L. del Mercato</searchLink><br /><searchLink fieldCode="AR" term="%22Gian+Pietro+Di+Sansebastiano%22">Gian Pietro Di Sansebastiano</searchLink>
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  Data: Plants, Vol 13, Iss 24, p 3530 (2024)
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  Data: MDPI AG, 2024.
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  Data: 2024
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  Data: LCC:Botany
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  Data: <searchLink fieldCode="DE" term="%22Viola+ucriana%22">Viola ucriana</searchLink><br /><searchLink fieldCode="DE" term="%22ex+situ+conservation%22">ex situ conservation</searchLink><br /><searchLink fieldCode="DE" term="%22Temporary+Immersion+System+%28TIS%29%22">Temporary Immersion System (TIS)</searchLink><br /><searchLink fieldCode="DE" term="%22flow+cytometry%22">flow cytometry</searchLink><br /><searchLink fieldCode="DE" term="%22Botany%22">Botany</searchLink><br /><searchLink fieldCode="DE" term="%22QK1-989%22">QK1-989</searchLink>
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  Label: Description
  Group: Ab
  Data: Viola ucriana Erben & 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® 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® 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.
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  Data: https://www.mdpi.com/2223-7747/13/24/3530; https://doaj.org/toc/2223-7747
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      – SubjectFull: Viola ucriana
        Type: general
      – SubjectFull: ex situ conservation
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      – SubjectFull: Temporary Immersion System (TIS)
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