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The Ca2+ Channel CYCLIC NUCLEOTIDE GATED CHANNEL13 (CNGC13) regulates systemic wound signalling and immunity against Spodoptera herbivory

Prajapati, R.; Kumari, M.; Singh, Y.; Pawar, M.; Marchesano, B. M. O.; Kalia, J.; Costa, A.; Vadassery, J.

2025-06-17 plant biology
10.1101/2025.06.16.660006 bioRxiv
Show abstract

Insect herbivory elicits rapid and complex signalling responses in plants, including membrane depolarization, cytosolic Ca2+ elevation and activation of jasmonate-mediated defense pathways in local and systemic leaves. These early events are orchestrated by calcium-permeable channels and regulatory components that generate stimulus-specific cytosolic Ca2+ signatures. However, the molecular mechanisms that generate long-distance Ca2+ propagation and systemic immune activation on herbivory remain poorly defined. Here, we identify CYCLIC NUCLEOTIDE GATED CHANNEL13 (CNGC13) as a novel plasma membrane-localized, phloem-expressed Ca2+ channel that is essential for systemic wound signalling and defense against lepidopteran pest, Spodoptera litura in Arabidopsis thaliana. CNGC13 is rapidly induced by wounding and herbivory, mediates cytosolic Ca2+ influx, and is required for efficient propagation of Ca2+ signals from wounded to distal leaves. Loss of CNGC13 compromises the thioglucosidase-dependent breakdown of aliphatic glucosinolates into aglucones/isothiocyanates within the vascular tissue, which are essential for wounding-induced systemic Ca2+ wave propagation. Consequently, cngc13 mutants display impaired systemic jasmonate accumulation in distal leaves, reduced glucosinolate levels and enhanced herbivore susceptibility. CNGC13 is also required for AtPep-induced Ca2+ elevation and Reactive Oxygen Species (ROS) signalling and interacts with the receptor kinase, PEPR2, implicating it in DAMP-mediated immune responses. Although CNGC13 and CNGC19 function cooperatively in herbivore defense, they do not physically interact, suggesting independent but converging roles. Our study identifies CNGC13 as a vasculature-localized Ca2+-permeable channel that functions downstream of PEPR2 receptor, to sustain systemic signalling, operating in a distinct yet converging pathway alongside known glutamate receptor-like channels during long-distance wound responses. SignificancePlants must detect and respond to insect herbivory rapidly by activating systemic immunity to trigger defense responses. Calcium (Ca2+) signalling plays a central role in this defense network, yet the identities and specific functions of Ca2+ channels involved in long-distance herbivory signalling remain poorly understood. Here, we identify the plasma membrane-localized channel CYCLIC NUCLEOTIDE GATED CHANNEL13 (CNGC13) as a key integrator of mechanical damage perception, ricca factor-mediated systemic signalling, and jasmonate-dependent defense against Spodoptera herbivory in Arabidopsis. CNGC13 facilitates systemic Ca2+ wave propagation, activates jasmonate and glucosinolate biosynthesis, and promotes the perception of damage-associated molecular pattern (DAMP) molecules to confer herbivore resistance. This work uncovers a previously uncharacterized calcium channel that plays a crucial dual role in systemic wound signalling.

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