Mediated Transformation of Tamarillo (<i>Solanum betaceum</i>) Callus Cell Suspension Cultures: A Novel Platform for Biotechnological Applications
<i>Solanum betaceum</i> Cav. (tamarillo) has a strong biotechnological potential given the ease of obtaining cell lines from it that can be genetically transformed. However, genetic transformation of tamarillo cell suspension cultures has not yet been described. This study presents a sim...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-03-01
|
| Series: | Plants |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2223-7747/14/7/1028 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850212851927482368 |
|---|---|
| author | Ricardo Ferraz Bruno Casimiro Daniela Cordeiro Jorge Canhoto Sandra Correia |
| author_facet | Ricardo Ferraz Bruno Casimiro Daniela Cordeiro Jorge Canhoto Sandra Correia |
| author_sort | Ricardo Ferraz |
| collection | DOAJ |
| description | <i>Solanum betaceum</i> Cav. (tamarillo) has a strong biotechnological potential given the ease of obtaining cell lines from it that can be genetically transformed. However, genetic transformation of tamarillo cell suspension cultures has not yet been described. This study presents a simple method for <i>Agrobacterium</i>-mediated transformation of these cells and demonstrates the successful insertion of the β-glucuronidase gene (<i>gusA</i>) and the yellow fluorescent protein gene (<i>eyfp</i>) in their genome. For the success of this protocol, the selection of actively growing sub-cultured callus as explant and isolation of bacterial colonies with a cell density OD<sub>600</sub> of 0.6–0.8 were key steps. Also, the inoculation of the callus in a bacteria liquid culture, the use of sonication, and the addition of antioxidants were essential. The transient expression of the <i>gusA</i> gene in tamarillo callus was confirmed and quantified, and no significant differences were observed between using LBA4404 or EHA105 strains. Finally, the insertion of the <i>eyfp</i> gene in the tamarillo genome enabled the in vivo confirmation of the transformation success. The present study showed that tamarillo cell suspension cultures can be genetically modified, opening the way for metabolite production in transformed cells and future scaling-up in bioreactors. |
| format | Article |
| id | doaj-art-6167483432ee4524b6545c80fa51119f |
| institution | OA Journals |
| issn | 2223-7747 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj-art-6167483432ee4524b6545c80fa51119f2025-08-20T02:09:14ZengMDPI AGPlants2223-77472025-03-01147102810.3390/plants14071028Mediated Transformation of Tamarillo (<i>Solanum betaceum</i>) Callus Cell Suspension Cultures: A Novel Platform for Biotechnological ApplicationsRicardo Ferraz0Bruno Casimiro1Daniela Cordeiro2Jorge Canhoto3Sandra Correia4Centre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, PortugalCentre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, PortugalCentre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, PortugalCentre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, PortugalCentre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, Portugal<i>Solanum betaceum</i> Cav. (tamarillo) has a strong biotechnological potential given the ease of obtaining cell lines from it that can be genetically transformed. However, genetic transformation of tamarillo cell suspension cultures has not yet been described. This study presents a simple method for <i>Agrobacterium</i>-mediated transformation of these cells and demonstrates the successful insertion of the β-glucuronidase gene (<i>gusA</i>) and the yellow fluorescent protein gene (<i>eyfp</i>) in their genome. For the success of this protocol, the selection of actively growing sub-cultured callus as explant and isolation of bacterial colonies with a cell density OD<sub>600</sub> of 0.6–0.8 were key steps. Also, the inoculation of the callus in a bacteria liquid culture, the use of sonication, and the addition of antioxidants were essential. The transient expression of the <i>gusA</i> gene in tamarillo callus was confirmed and quantified, and no significant differences were observed between using LBA4404 or EHA105 strains. Finally, the insertion of the <i>eyfp</i> gene in the tamarillo genome enabled the in vivo confirmation of the transformation success. The present study showed that tamarillo cell suspension cultures can be genetically modified, opening the way for metabolite production in transformed cells and future scaling-up in bioreactors.https://www.mdpi.com/2223-7747/14/7/1028cell suspension culturesGUS activity assayin vivo monitoringrelative qPCRSolanaceae |
| spellingShingle | Ricardo Ferraz Bruno Casimiro Daniela Cordeiro Jorge Canhoto Sandra Correia Mediated Transformation of Tamarillo (<i>Solanum betaceum</i>) Callus Cell Suspension Cultures: A Novel Platform for Biotechnological Applications Plants cell suspension cultures GUS activity assay in vivo monitoring relative qPCR Solanaceae |
| title | Mediated Transformation of Tamarillo (<i>Solanum betaceum</i>) Callus Cell Suspension Cultures: A Novel Platform for Biotechnological Applications |
| title_full | Mediated Transformation of Tamarillo (<i>Solanum betaceum</i>) Callus Cell Suspension Cultures: A Novel Platform for Biotechnological Applications |
| title_fullStr | Mediated Transformation of Tamarillo (<i>Solanum betaceum</i>) Callus Cell Suspension Cultures: A Novel Platform for Biotechnological Applications |
| title_full_unstemmed | Mediated Transformation of Tamarillo (<i>Solanum betaceum</i>) Callus Cell Suspension Cultures: A Novel Platform for Biotechnological Applications |
| title_short | Mediated Transformation of Tamarillo (<i>Solanum betaceum</i>) Callus Cell Suspension Cultures: A Novel Platform for Biotechnological Applications |
| title_sort | mediated transformation of tamarillo i solanum betaceum i callus cell suspension cultures a novel platform for biotechnological applications |
| topic | cell suspension cultures GUS activity assay in vivo monitoring relative qPCR Solanaceae |
| url | https://www.mdpi.com/2223-7747/14/7/1028 |
| work_keys_str_mv | AT ricardoferraz mediatedtransformationoftamarilloisolanumbetaceumicalluscellsuspensionculturesanovelplatformforbiotechnologicalapplications AT brunocasimiro mediatedtransformationoftamarilloisolanumbetaceumicalluscellsuspensionculturesanovelplatformforbiotechnologicalapplications AT danielacordeiro mediatedtransformationoftamarilloisolanumbetaceumicalluscellsuspensionculturesanovelplatformforbiotechnologicalapplications AT jorgecanhoto mediatedtransformationoftamarilloisolanumbetaceumicalluscellsuspensionculturesanovelplatformforbiotechnologicalapplications AT sandracorreia mediatedtransformationoftamarilloisolanumbetaceumicalluscellsuspensionculturesanovelplatformforbiotechnologicalapplications |