Development of an efficient protoplast isolation and transfection system for castor bean ( Ricinus communis L.)
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ORIGINAL ARTICLE
Development of an efficient protoplast isolation and transfection system for castor bean (Ricinus communis L.) Liang Bai2,3 · Yan Cheng2,3 · Jikai She2 · Zhibiao He3,4 · Haiping Liu1 · Guowen Zhang2 · Ruizhen Cao2 · Yongsheng Chen1,3 Received: 5 March 2020 / Accepted: 12 September 2020 © Springer Nature B.V. 2020
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. Keywords Ricinus communis L. · PEG-mediated transfection · Protoplast · Gene expression
Introduction Protoplasts are plant cells that have had the cell wall removed, but they retain the main cell features (Davey et al. 2005). Like eukaryotic cells, they are a versatile model system for observing various cellular events. Protoplasts are Communicated by Maria Margarida Oliveira. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11240-020-01932-0) contains supplementary material, which is available to authorized users. * Yongsheng Chen [email protected] 1
College of Life Sciences and Food Engineering, Inner Mongolia University for Nationalities, Tongliao 028000, China
2
College of Medicine, Inner Mongolia University for Nationalities, Tongliao 028000, China
3
Inner Mongolia Industrial Engineering Research Center of Universities for Castor, Tongliao 028000, China
4
Tongliao Academy of Agricultural Science, Tongliao 028000, Inner Mongolia, China
an effective tool for demonstrating plant cell hybridization and regeneration (Wenck and Marton 1995), especially when used as a versatile cell-based system for genetic research (Krens et al. 1982). In addition, protoplasts can maintain the physiological and cellular processes of whole plants (Sheen 2001). Hence, protoplasts offer fundamental value for contributing to protein subcellular localization (Yoo et al. 2007), protein–protein interaction (Ehlert et al. 2006), signal transduction (Kanofsky et al. 2019), promoter screening (Sultana et a
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