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

BMC Research Notes

Research Note

Carsten Harms¹  Danna Susunaga-Gómez²  Paula Figueroa-Varela²  Catalina Restrepo-Osorio²  Diego Villanueva-Mejía² 

[1] Bremerhaven Institute for Applied Molecular Biology, University of Applied Sciences Bremerhaven, Bremerhaven, Germany;CIBIOP Research Group, School of Applied Sciences and Engineering, EAFIT University, Medellín, Colombia;

关键词: Protoplasts;    Cellulase;    Macerozyme;    PEG-mediated transfection;   

DOI  :  10.1186/s13104-023-06414-y

 received in 2022-09-25, accepted in 2023-06-20,  发布年份 2023

来源: Springer

【 摘 要 】

ObjectiveThe 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.ResultsFactors 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.

【 授权许可】

CC BY   
© The Author(s) 2023

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