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

Yunjing Wang; Qian Gong; Fan Huang; Linfang He; Yule Liu

doi:10.1016/j.xpro.2021.101040

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

STAR Protoc. 2021 Dec 15;3(1):101040. doi: 10.1016/j.xpro.2021.101040. eCollection 2022 Mar 18.

Authors

Yunjing Wang^{1

2}, Qian Gong^{1

2}, Fan Huang^{1

2}, Linfang He^{1

2}, Yule Liu^{1

2}

Affiliations

¹ MOE Key Laboratory of Bioinformatics and Center for Plant Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
² Tsinghua-Peking Center for Life Sciences, Beijing 100084, China.

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).

Keywords: Microscopy; Model Organisms; Plant sciences; Signal Transduction.

Publication types

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

MeSH terms

Animals
Arabidopsis* / metabolism
Calcium / metabolism
Calcium Signaling
Insecta / metabolism
Nicotiana* / metabolism

Substances

Calcium