Transient gene expression system in zoysiagrass leaf mesophyll protoplasts

被引:0
作者
Jin Hee Kim
Phan Phuong Thao Doan
Hyo-Yeon Lee
Jeongsik Kim
机构
[1] Subtropical Horticulture Research Institute,Interdisciplinary Graduate Program in Advanced Convergence Technology and Science
[2] Jeju National University,Department of Biotechnology
[3] Jeju National University,Faculty of Science Education
[4] Jeju National University,undefined
[5] Jeju National University,undefined
来源
Plant Biotechnology Reports | 2022年 / 16卷
关键词
Protoplast; Transient expression; Polyethylene glycol; Zoysiagrass;
D O I
暂无
中图分类号
学科分类号
摘要
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.
引用
收藏
页码:113 / 121
页数:8
相关论文
共 50 条
  • [31] An efficient PEG-mediated transient gene expression system in grape protoplasts and its application in subcellular localization studies of flavonoids biosynthesis enzymes
    Wang, Huiling
    Wang, Wei
    Zhan, Jicheng
    Huang, Weidong
    Xu, Haiying
    [J]. SCIENTIA HORTICULTURAE, 2015, 191 : 82 - 89
  • [32] Establishment of protoplasts isolation and transient transformation system for kiwifruit
    Zhang, Xiaoli
    Peng, Rui
    Tian, Xinbo
    Guo, Yuqi
    Li, Xinyi
    Liu, Xinling
    Xie, Yue
    Li, Mingzhang
    Xia, Hui
    Liang, Dong
    [J]. SCIENTIA HORTICULTURAE, 2024, 329
  • [33] TRANSIENT FOREIGN GENE-EXPRESSION IN POLYETHYLENE-GLYCOL TREATED OR ELECTROPULSATED EUCALYPTUS-GUNNII PROTOPLASTS
    TEULIERES, C
    GRIMAPETTENATI, J
    CURIE, C
    TEISSIE, J
    BOUDET, AM
    [J]. PLANT CELL TISSUE AND ORGAN CULTURE, 1991, 25 (02) : 125 - 132
  • [34] LENTIL ROOT PROTOPLASTS - A TRANSIENT EXPRESSION SYSTEM SUITABLE FOR COELECTROPORATION OF MONOCLONAL-ANTIBODIES AND PLASMID MOLECULES
    MACCARRONE, M
    VELDINK, GA
    AGRO, AF
    VLIEGENTHART, JFG
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 1995, 1243 (01): : 136 - 142
  • [35] Establishment of protoplasts transient expression system in Pinellia ternata (Thunb.) Breit
    Yu-hang Tian
    Miao Liu
    Liu Tang
    Yu-jin Zhang
    Ye Hang
    Li-yang Shangguan
    Yin-qun Zhang
    Ming-sheng Zhang
    [J]. Biotechnology Letters, 2023, 45 : 1381 - 1391
  • [36] Establishment of protoplasts transient expression system in Pinellia ternata (Thunb.) Breit
    Tian, Yu-hang
    Liu, Miao
    Tang, Liu
    Zhang, Yu-jin
    Hang, Ye
    Shangguan, Li-yang
    Zhang, Yin-qun
    Zhang, Ming-sheng
    [J]. BIOTECHNOLOGY LETTERS, 2023, 45 (10) : 1381 - 1391
  • [37] An efficient mesophyll protoplast isolation, purification and PEG-mediated transient gene expression for subcellular localization in Chinese kale
    Sun, Bo
    Zhang, Fen
    Xiao, Nan
    Jiang, Min
    Yuan, Qiao
    Xue, Shengling
    Miao, Huiying
    Chen, Qing
    Li, Mengyao
    Wang, Xiaorong
    Wang, Qiaomei
    Tang, Haoru
    [J]. SCIENTIA HORTICULTURAE, 2018, 241 : 187 - 193
  • [38] Plant regeneration via somatic embryogenesis, and transient gene expression in sweet potato protoplasts
    Dhir, SK
    Oglesby, J
    Bhagsari, AS
    [J]. PLANT CELL REPORTS, 1998, 17 (09) : 665 - 669
  • [39] Plant regeneration via somatic embryogenesis, and transient gene expression in sweet potato protoplasts
    S. K. Dhir
    J. Oglesby
    A. S. Bhagsari
    [J]. Plant Cell Reports, 1998, 17 : 665 - 669
  • [40] Transient expression of UIDA gene in grapevine protoplasts after PEG-mediated transformation
    Jardak, R
    Mliki, A
    Ghorbel, A
    Reustle, GM
    [J]. JOURNAL INTERNATIONAL DES SCIENCES DE LA VIGNE ET DU VIN, 2002, 36 (02): : 93 - 95