Preparation of epidermal peels and guard cell protoplasts for cellular, Electrophysiological, and -omics assays of guard cell function

Mengmeng Zhu, Byeong Wook Jeon, Sisi Geng, Yunqing Yu, Kelly Balmant, Sixue Chen, Sarah Mary Assmann

Research output: Chapter in Book/Report/Conference proceedingChapter

12 Scopus citations

Abstract

Bioassays are commonly used to study stomatal phenotypes. There are multiple options in the choice of plant materials and species used for observation of stomatal and guard cell responses in vivo. Here, detailed procedures for bioassays of stomatal responses to abscisic acid (ABA) in Arabidopsis thaliana are described, including ABA promotion of stomatal closure, ABA inhibition of stomatal opening, and ABA promotion of reaction oxygen species (ROS) production in guard cells. We also include an example of a stomatal bioassay for the guard cell CO2 response using guard cell-enriched epidermal peels from Brassica napus. Highly pure preparations of guard cell protoplasts can be produced, which are also suitable for studies on guard cell signaling, as well as for studies on guard cell ion transport. Small-scale and large-scale guard cell protoplast preparations are commonly used for electrophysiological and -omics studies, respectively. We provide a procedure for small-scale guard cell protoplasting from A. thaliana. Additionally, a general protocol for large-scale preparation of guard cell protoplasts, with specifications for three different species, A. thaliana, B. napus, and Vicia faba is also provided.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages89-121
Number of pages33
DOIs
StatePublished - Jan 1 2016

Publication series

NameMethods in Molecular Biology
Volume1363
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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    Zhu, M., Jeon, B. W., Geng, S., Yu, Y., Balmant, K., Chen, S., & Assmann, S. M. (2016). Preparation of epidermal peels and guard cell protoplasts for cellular, Electrophysiological, and -omics assays of guard cell function. In Methods in Molecular Biology (pp. 89-121). (Methods in Molecular Biology; Vol. 1363). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3115-6_9