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Development of an efficient protoplast isolation and transfection system for castor bean (Ricinus communis L.)

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Abstract

Castor bean is an oil crop plant (Euphorbiaceae) found across the tropical, subtropical, and temperate regions. Despite its important oil properties and cultivation in a wide range of environments, the molecular mechanisms of castor’s adaptation and metabolism have not been fully clarified due to difficulties in genetic modification approaches. The protoplasts of several other plant species have been used as versatile cell-based model systems to elucidate the biological functions of genes and proteins. Here, we report an optimized protocol for protoplast isolation from the leaves and cotyledons of castor bean. The main parameters evaluated to achieve the maximum protoplast yield were the application of a cell wall-degrading enzyme solution, the osmotic pressure of the enzymolysis solution, and the enzymolysis time. Transient expression and the main influencing factors were validated by fluorescence microscopy of castor protoplasts. Our results suggest that castor protoplasts can be used as a productive cell-based system to explore the mechanisms involved in the molecular, biochemical, and functional characterization of castor bean genes.

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Acknowledgements

This work was supported by the Natural Science Foundation of China (Grant Nos. 31460353), the Natural Science Foundation of Inner Mongolia (Grant Nos. 2017MS0340), and the Natural Scientific Foundation of Inner Mongolia University for Nationalities (Grant Nos. NMDGP17102). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. College of Life Sciences and Food Engineering, Inner Mongolia University for Nationalities, Tongliao, 028000, China

    Haiping Liu & Yongsheng Chen

  2. College of Medicine, Inner Mongolia University for Nationalities, Tongliao, 028000, China

    Liang Bai, Yan Cheng, Jikai She, Guowen Zhang & Ruizhen Cao

  3. Inner Mongolia Industrial Engineering Research Center of Universities for Castor, Tongliao, 028000, China

    Liang Bai, Yan Cheng, Zhibiao He & Yongsheng Chen

  4. Tongliao Academy of Agricultural Science, Tongliao, 028000, Inner Mongolia, China

    Zhibiao He

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  1. Liang Bai
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Contributions

LB and YC carried out the experimental design. LB, JS, GZ, RC, and WQ performed the experiments. KL contributed reagents and analytical tools. AM and YC analysed the data. BL wrote the manuscript. All authors read and approved the manuscript.

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Correspondence to Yongsheng Chen.

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Communicated by Maria Margarida Oliveira.

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Bai, L., Cheng, Y., She, J. et al. Development of an efficient protoplast isolation and transfection system for castor bean (Ricinus communis L.). Plant Cell Tiss Organ Cult 143, 457–464 (2020). https://doi.org/10.1007/s11240-020-01932-0

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