Abstract
Decoding wound signalling in plants is critical for understanding various aspects of plant sciences, from pest resistance to secondary metabolite and phytohormone biosynthesis. The plant defence responses are known to primarily involve NADPH-oxidase-mediated H2O2 and Ca2+ signalling pathways, which propagate across long distances through the plant vasculature and tissues. Using non-destructive optical nanosensors, we find that the H2O2 concentration profile post-wounding follows a logistic waveform for six plant species: lettuce (Lactuca sativa), arugula (Eruca sativa), spinach (Spinacia oleracea), strawberry blite (Blitum capitatum), sorrel (Rumex acetosa) and Arabidopsis thaliana, ranked in order of wave speed from 0.44 to 3.10 cm min−1. The H2O2 wave tracks the concomitant surface potential wave measured electrochemically. We show that the plant RbohD glutamate-receptor-like channels (GLR3.3 and GLR3.6) are all critical to the propagation of the wound-induced H2O2 wave. Our findings highlight the utility of a new type of nanosensor probe that is species-independent and capable of real-time, spatial and temporal biochemical measurements in plants.
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The authors declare that all data supporting the findings of this study are available within the paper and any raw data can be obtained from the corresponding author on request.
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Acknowledgements
We thank E. Farmer for atglr3.3 atglr3.6 seeds. This research was supported by the National Research Foundation (NRF), Prime Minister’s Office, Singapore under its Campus for Research Excellence and Technological Enterprise (CREATE) program. The Disruptive & Sustainable Technology for Agricultural Precision (DiSTAP) is an interdisciplinary research group of the Singapore MIT Alliance for Research and Technology (SMART) Centre. T.T.S.L. was supported on a graduate fellowship by the Agency of Science, Research and Technology, Singapore. K.S.S. was supported by the Department of Energy Computational Science Graduate Fellowship program under grant DE-FG02-97ER25308. M.P. is grateful for the support of the Samsung scholarship.
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T.T.S.L., N.-H.C. and M.S.S. conceived the project, designed the study and wrote the manuscript. T.T.S.L. performed the majority of experiments and data analysis. J.S.S. provided A. thaliana mutants and helped with result discussions. V.B.K. performed the electrical signal measurements. K.S.S. assisted with the numerical simulation. P.G. performed AFM analysis. S.-Y.K., M.P., M.C.-Y.A., K.D.T., M.A.L. and M.B.C.-P. assisted with the experimental design and discussions. All authors have revised the manuscript and given their approval of the final version.
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Supplementary Figures 1–18, Supplementary Discussion, Supplementary Table 1 and Supplementary Video legends.
Supplementary Video 1
False-coloured video of nanosensors’ response towards wounding in a spinach leaf.
Supplementary Video 2
Real-time response of nanosensors upon insect feeding on a spinach leaf.
Supplementary Video 3
Spatiotemporal profile of G- SWNT sensor response due to wounding.
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Lew, T.T.S., Koman, V.B., Silmore, K.S. et al. Real-time detection of wound-induced H2O2 signalling waves in plants with optical nanosensors. Nat. Plants 6, 404–415 (2020). https://doi.org/10.1038/s41477-020-0632-4
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DOI: https://doi.org/10.1038/s41477-020-0632-4
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