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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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Detalles Bibliográficos
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
Descripción
Sumario: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.