In JoVE (1)
Other Publications (1)
Articles by James Canham in JoVE
Real-time In Vivo Recording of Arabidopsis Calcium Signals During Insect Feeding Using a Fluorescent Biosensor Thomas R. Vincent1, James Canham1, Masatsugu Toyota2,3,4, Marieta Avramova1, Sam T. Mugford5, Simon Gilroy2, Anthony J. Miller1, Saskia Hogenhout5, Dale Sanders1 1Department of Metabolic Biology, John Innes Centre, Norwich Research Park, 2Department of Botany, University of Wisconsin, Madison, 3Department of Biochemistry and Molecular Biology, Saitama University, 4Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), 5Department of Cell and Developmental Biology, John Innes Centre, Norwich Research Park This protocol outlines a simple method for analyzing calcium signals in plants generated by feeding hemipteran insects, such as aphids. Arabidopsis thaliana transformed with the GFP calcium biosensor GCaMP3 allow for the real-time in vivo imaging of calcium dynamics with a high temporal and spatial resolution.
Other articles by James Canham on PubMed
Interplay of Plasma Membrane and Vacuolar Ion Channels, Together with BAK1, Elicits Rapid Cytosolic Calcium Elevations in Arabidopsis During Aphid Feeding The Plant Cell. Jun, 2017 | Pubmed ID: 28559475 A transient rise in cytosolic calcium ion concentration is one of the main signals used by plants in perception of their environment. The role of calcium in the detection of abiotic stress is well documented; however, its role during biotic interactions remains unclear. Here, we use a fluorescent calcium biosensor (GCaMP3) in combination with the green peach aphid (Myzus persicae) as a tool to study Arabidopsis thaliana calcium dynamics in vivo and in real time during a live biotic interaction. We demonstrate rapid and highly localized plant calcium elevations around the feeding sites of M. persicae, and by monitoring aphid feeding behavior electrophysiologically, we demonstrate that these elevations correlate with aphid probing of epidermal and mesophyll cells. Furthermore, we dissect the molecular mechanisms involved, showing that interplay between the plant defense coreceptor BRASSINOSTEROID INSENSITIVE-ASSOCIATED KINASE1 (BAK1), the plasma membrane ion channels GLUTAMATE RECEPTOR-LIKE 3.3 and 3.6 (GLR3.3 and GLR3.6), and the vacuolar ion channel TWO-PORE CHANNEL1 (TPC1) mediate these calcium elevations. Consequently, we identify a link between plant perception of biotic threats by BAK1, cellular calcium entry mediated by GLRs, and intracellular calcium release by TPC1 during a biologically relevant interaction.