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

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).