An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines
Abstract 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...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-07-01
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| Series: | BMC Research Notes |
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| Online Access: | https://doi.org/10.1186/s13104-023-06414-y |
| _version_ | 1827905711796912128 |
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| author | Paula Figueroa-Varela Danna Susunaga-Gómez Catalina Restrepo-Osorio Carsten Harms Diego Villanueva-Mejía |
| author_facet | Paula Figueroa-Varela Danna Susunaga-Gómez Catalina Restrepo-Osorio Carsten Harms Diego Villanueva-Mejía |
| author_sort | Paula Figueroa-Varela |
| collection | DOAJ |
| description | Abstract 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 × 104 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 × 105 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. |
| first_indexed | 2024-03-13T00:45:05Z |
| format | Article |
| id | doaj.art-fa260c904efc4daa98cd1393ea7a3570 |
| institution | Directory Open Access Journal |
| issn | 1756-0500 |
| language | English |
| last_indexed | 2024-03-13T00:45:05Z |
| publishDate | 2023-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Research Notes |
| spelling | doaj.art-fa260c904efc4daa98cd1393ea7a35702023-07-09T11:05:16ZengBMCBMC Research Notes1756-05002023-07-011611810.1186/s13104-023-06414-yAn efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) linesPaula Figueroa-Varela0Danna Susunaga-Gómez1Catalina Restrepo-Osorio2Carsten Harms3Diego Villanueva-Mejía4CIBIOP Research Group, School of Applied Sciences and Engineering, EAFIT UniversityCIBIOP Research Group, School of Applied Sciences and Engineering, EAFIT UniversityCIBIOP Research Group, School of Applied Sciences and Engineering, EAFIT UniversityBremerhaven Institute for Applied Molecular Biology, University of Applied Sciences BremerhavenCIBIOP Research Group, School of Applied Sciences and Engineering, EAFIT UniversityAbstract 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 × 104 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 × 105 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.https://doi.org/10.1186/s13104-023-06414-yProtoplastsCellulaseMacerozymePEG-mediated transfection |
| spellingShingle | Paula Figueroa-Varela Danna Susunaga-Gómez Catalina Restrepo-Osorio Carsten Harms Diego Villanueva-Mejía An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines BMC Research Notes Protoplasts Cellulase Macerozyme PEG-mediated transfection |
| 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 | Protoplasts Cellulase Macerozyme PEG-mediated transfection |
| url | https://doi.org/10.1186/s13104-023-06414-y |
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