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An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines

OBJECTIVE: The purpose of this study was to develop a method for the isolation, culture, and PEG-mediated protoplast transfection from leaves of in vitro-grown plants of Ricinus communis. RESULTS: Factors such as the enzymatic composition and the incubation time were evaluated. The enzymatic solutio...

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Autores principales: Figueroa-Varela, Paula, Susunaga-Gómez, Danna, Restrepo-Osorio, Catalina, Harms, Carsten, Villanueva-Mejía, Diego
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10327310/
https://www.ncbi.nlm.nih.gov/pubmed/37415245
http://dx.doi.org/10.1186/s13104-023-06414-y
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author Figueroa-Varela, Paula
Susunaga-Gómez, Danna
Restrepo-Osorio, Catalina
Harms, Carsten
Villanueva-Mejía, Diego
author_facet Figueroa-Varela, Paula
Susunaga-Gómez, Danna
Restrepo-Osorio, Catalina
Harms, Carsten
Villanueva-Mejía, Diego
author_sort Figueroa-Varela, Paula
collection PubMed
description OBJECTIVE: The purpose of this study was to develop a method for the isolation, culture, and PEG-mediated protoplast transfection from leaves of in vitro-grown plants of Ricinus communis. RESULTS: Factors such as the enzymatic composition and the incubation time were evaluated. The enzymatic solution, containing 1.6% Cellulase-R10 and 0.8% Macerozyme-R10, with 16 h of incubation, was the best condition to achieve a high protoplast yield (481.16 × 10(4) protoplasts/g FW) with a high percentage of viability (95%). The combination and concentration of enzymes have been shown to affect the protoplast isolation efficiency significantly. Furthermore, we found that a higher number of protoplasts (8.5 × 10(5) protoplast/g FW) was obtained at a longer incubation time, but their viability decreased. We obtained a simple and efficient protocol to isolate protoplast from Ricinus communis leaves and culture. A PEG-mediated protoplast transfection protocol was also established to introduce plasmid DNA into Ricinus communis genotypes cultivated in Colombia. Thus, strengthening advances in the genetic improvement processes for this crop are presented. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13104-023-06414-y.
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spelling pubmed-103273102023-07-08 An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines Figueroa-Varela, Paula Susunaga-Gómez, Danna Restrepo-Osorio, Catalina Harms, Carsten Villanueva-Mejía, Diego BMC Res Notes Research Note OBJECTIVE: The purpose of this study was to develop a method for the isolation, culture, and PEG-mediated protoplast transfection from leaves of in vitro-grown plants of Ricinus communis. RESULTS: Factors such as the enzymatic composition and the incubation time were evaluated. The enzymatic solution, containing 1.6% Cellulase-R10 and 0.8% Macerozyme-R10, with 16 h of incubation, was the best condition to achieve a high protoplast yield (481.16 × 10(4) protoplasts/g FW) with a high percentage of viability (95%). The combination and concentration of enzymes have been shown to affect the protoplast isolation efficiency significantly. Furthermore, we found that a higher number of protoplasts (8.5 × 10(5) protoplast/g FW) was obtained at a longer incubation time, but their viability decreased. We obtained a simple and efficient protocol to isolate protoplast from Ricinus communis leaves and culture. A PEG-mediated protoplast transfection protocol was also established to introduce plasmid DNA into Ricinus communis genotypes cultivated in Colombia. Thus, strengthening advances in the genetic improvement processes for this crop are presented. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13104-023-06414-y. BioMed Central 2023-07-06 /pmc/articles/PMC10327310/ /pubmed/37415245 http://dx.doi.org/10.1186/s13104-023-06414-y Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Note
Figueroa-Varela, Paula
Susunaga-Gómez, Danna
Restrepo-Osorio, Catalina
Harms, Carsten
Villanueva-Mejía, Diego
An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines
title An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines
title_full An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines
title_fullStr An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines
title_full_unstemmed An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines
title_short An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines
title_sort efficient method for protoplast-mediated production of transformed castor bean (ricinus communis) lines
topic Research Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10327310/
https://www.ncbi.nlm.nih.gov/pubmed/37415245
http://dx.doi.org/10.1186/s13104-023-06414-y
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