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Live imaging and quantitation of insect feeding-induced Ca2+ signal using GCaMP3-based system in Nicotiana benthamiana
- Source :
- STAR Protocols, Vol 3, Iss 1, Pp 101040- (2022)
- Publication Year :
- 2022
- Publisher :
- Elsevier, 2022.
-
Abstract
- Summary: 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).
Details
- Language :
- English
- ISSN :
- 26661667
- Volume :
- 3
- Issue :
- 1
- Database :
- Directory of Open Access Journals
- Journal :
- STAR Protocols
- Publication Type :
- Academic Journal
- Accession number :
- edsdoj.506431804c8141288bbd9635a113109e
- Document Type :
- article
- Full Text :
- https://doi.org/10.1016/j.xpro.2021.101040