Live imaging and quantitation of insect feeding-induced Ca²⁺ signal using GCaMP3-based system in Nicotiana benthamiana.

ABSTRACT: Wounding evokes transient increases in cytosolic calcium (Ca²⁺) concentration. Visualizing real-time Ca²⁺ flux provides new insights into Ca²⁺-signaling pathways. Here, we outline a protocol to detect insect feeding-induced Ca²⁺ flux elevation in Nicotiana benthamiana leaves based on the GCaMP3 reporter system by Leica fluorescence stereo microscopes (LFSM). LFSM combines super-fast manual screening with high-end imaging capabilities. Through this protocol, we can clearly observe the calcium flow after aphid's piercing-sucking. Additionally, we describe a protocol to quantify Ca²⁺ level using LFSM. For complete details on the use and execution of this protocol, please refer to Wang et al. (2021).

SUBMITTER: Wang Y 

PROVIDER: S-EPMC8689350 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature