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Transient gene expression system in zoysiagrass leaf mesophyll protoplasts

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Abstract

Transient expression of genes in protoplasts is a versatile technique for rapid functional characterization of genes by assessing protein localization or effector–reporter responses. In addition, protoplasts have been widely used for generating transgenic or gene-edited plants in model or crop plants. Zoysiagrass (Zoysia japonica Steud.) is one of the most economically important turf plants used in many living places or natural fields, but its management is labor-intensive. Therefore, genetic manipulation using valuable genes is highly demanded in zoysiagrass. Although transient expression systems in zoysiagrass facilitates the identification of valuable zoysiagrass genes, transient expression use in the zoysiagrass protoplasts is yet to be reported. Here we describe the methodology and feasibility for transient expression of genes in the zoysiagrass protoplasts isolated from green leaves. We obtained more than 70% of transfection efficiency in the zoysiagrass protoplasts using polyethylene glycol-mediated transfection. Additionally, we showed the feasibility of cellular, biochemical, and molecular approaches using the zoysiagrass protoplasts transiently expressing genes of interest through fluorescent microscopic observation, immunoblot analysis, and luciferase-based promoter activity assay. Along with the genome draft of the Zoysia family, transient expression will be a valuable tool to rapidly investigate gene functions. An initial assessment of gene function using protoplasts facilitate the identification of valuable genes that can improve the economic value of the Z. japonica or its closely related species by gene manipulation.

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Acknowledgements

This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education (2019R1A6A1A11052070 and 2020R1I1A1A01057891) and Science & ICT (2018R1C1B5086056 and 2021R1A2C1012991). The grantors had not played any role in writing this report or the decision to submit this article for publication.

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Authors and Affiliations

  1. Subtropical Horticulture Research Institute, Jeju National University, Jeju, 63243, Republic of Korea

    Jin Hee Kim, Hyo-Yeon Lee & Jeongsik Kim

  2. Interdisciplinary Graduate Program in Advanced Convergence Technology and Science, Jeju National University, Jeju, 63243, Republic of Korea

    Phan Phuong Thao Doan & Jeongsik Kim

  3. Department of Biotechnology, Jeju National University, Jeju, 63243, Republic of Korea

    Hyo-Yeon Lee

  4. Faculty of Science Education, Jeju National University, Jeju, 63243, Republic of Korea

    Jeongsik Kim

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  1. Jin Hee Kim
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Kim, J.H., Doan, P.P.T., Lee, HY. et al. Transient gene expression system in zoysiagrass leaf mesophyll protoplasts. Plant Biotechnol Rep 16, 113–121 (2022). https://doi.org/10.1007/s11816-021-00726-w

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  • DOI: https://doi.org/10.1007/s11816-021-00726-w

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