Screening for Arabidopsis mutants with altered Ca2+ signal response using aequorin-based Ca2+ reporter system

Shujing Sun; Xiaoyan Zhang; Kong Chen; Xiaohong Zhu; Yang Zhao

doi:10.1016/j.xpro.2021.100558

Screening for Arabidopsis mutants with altered Ca²⁺ signal response using aequorin-based Ca²⁺ reporter system

STAR Protoc. 2021 May 23;2(2):100558. doi: 10.1016/j.xpro.2021.100558. eCollection 2021 Jun 18.

Authors

Shujing Sun¹, Xiaoyan Zhang^{2

3}, Kong Chen^{1

4}, Xiaohong Zhu^{2

3}, Yang Zhao^{1

2}

Affiliations

¹ Shanghai Center for Plant Stress Biology, and CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
² State Key Laboratory of Crop Stress Adaptation and Improvement, School of Life Sciences, Henan University, Kaifeng 475001, China.
³ State Key Laboratory of Cotton Biology, School of Life Sciences, Henan University, Kaifeng 475001, China.
⁴ University of Chinese Academy of Sciences, Beijing 100049, China.

Abstract

Environmental stimuli evoke transient increases of the cytosolic Ca²⁺ level. To identify upstream components of Ca²⁺ signaling, we have optimized two forward genetic screening systems based on Ca²⁺ reporter aequorin. AEQsig6 and AEQub plants were used for generating ethyl methanesulfonate (EMS)-mutagenized libraries. The AEQsig6 EMS-mutagenized library was preferably used to screen the mutants with reduced Ca²⁺ signal response due to its high effectiveness, while the AEQub EMS-mutagenized library was used for screening of the mutants with altered Ca²⁺ signal response. For complete details on the use and execution of this protocol, please refer to Chen et al. (2020) and Zhu et al. (2013).

Keywords: Genetics; High Throughput Screening; Model Organisms; Molecular/Chemical Probes; Plant sciences.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aequorin* / genetics
Aequorin* / metabolism
Arabidopsis / genetics*
Arabidopsis Proteins* / genetics
Arabidopsis Proteins* / metabolism
Calcium Signaling / genetics*
Gene Library
Luminescent Measurements
Mutation / genetics*
Whole Genome Sequencing

Substances

Arabidopsis Proteins
Aequorin