Unit

UNIT 2.8 Isolation, Culture, and Transient Transformation of Plant Protoplasts

  1. Jinbo Shen1,
  2. Jiaxin Fu3,
  3. Jin Ma4,
  4. Xiangfeng Wang1,
  5. Caiji Gao1,
  6. ChuXiong Zhuang3,
  7. Jianmin Wan4,
  8. Liwen Jiang1,2

Published Online: 3 JUN 2014

DOI: 10.1002/0471143030.cb0208s63

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Shen, J., Fu, J., Ma, J., Wang, X., Gao, C., Zhuang, C., Wan, J. and Jiang, L. 2014. Isolation, Culture, and Transient Transformation of Plant Protoplasts. Current Protocols in Cell Biology. 63:2.8:2.8.1–2.8.17.

Author Information

  1. 1

    School of Life Sciences, Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Hong Kong, China

  2. 2

    Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, China

  3. 3

    State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Sciences, South China Agricultural University, Guangzhou, China

  4. 4

    National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, China

Publication History

  1. Published Online: 3 JUN 2014

Abstract

Transient gene expression in protoplasts, which has been used in several plant species, is an important and versatile tool for rapid functional gene analysis, protein subcellular localization, and biochemical manipulations. This unit describes transient gene expression by electroporation of DNA into protoplasts of Arabidopsis or tobacco suspension-cultured cells and by polyethylene glycol (PEG)-mediated DNA transformation into protoplasts derived from rice leaf sheaths. PEG-mediated DNA transformation for transient gene expression in rice protoplasts in suspension culture is also described as an alternative technique. Methods for collecting intracellular and secreted proteins are also provided. Curr. Protoc. Cell Biol. 63:2.8.1-2.8.17. © 2014 by John Wiley & Sons, Inc.

Keywords:

  • Arabidopsis;
  • tobacco;
  • rice;
  • protoplast isolation;
  • transient expression;
  • electroporation;
  • PEG