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Acaricidal Mechanism of Scopoletin Against Tetranychus cinnabarinus.

ABSTRACT: Scopoletin is a promising acaricidal botanical natural compound against Tetranychus cinnabarinus, and its acaricidal mechanism maybe involve calcium overload according to our previous study. To seek potential candidate target genes of calcium overload induced by scopoletin in T. cinnabarinus, RNA-seq was utilized to detect changes in transcription levels. 24 and 48 h after treatment, 70 and 102 differentially expressed genes were obtained, respectively. Target genes included 3 signal transduction genes, 4 cell apoptosis genes, 4 energy metabolism genes, and 2 transcription factor genes. The role of 3 calcium signaling pathway-related genes, namely, G-protein-coupled neuropeptide receptor, Bcl-2 protein and guanylate kinase (designated TcGPCR, TcBAG, and TcGUK, respectively) in the calcium overload were investigated in this study. RT-qPCR detection showed that scopoletin treatment upregulated the expression level of TcGPCR and downregulated the expression level of TcBAG and TcGUK. The result of RNAi indicated that downregulation of TcGPCR decreased susceptibility to scopoletin, and downregulation of TcBAG and TcGUK enhanced susceptibility to scopoletin. Functional expression in Chinese hamster ovary cells showed that scopoletin induced a significant increase in intracellular free calcium [Ca2+]i levels by activating TcGPCR. These results demonstrated that the acaricidal mechanism of scopoletin was via disrupting intracellular Ca2+ homeostasis and calcium signaling pathway mediated by GPCR, BAG, and GUK.

SUBMITTER: Zhou H

PROVIDER: S-EPMC6414448 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Acaricidal Mechanism of Scopoletin Against Tetranychus cinnabarinus.

Zhou Hong H Zhang Yong-Qiang YQ Lai Ting T Liu Xue-Jiao XJ Guo Fu-You FY Guo Tao T Ding Wei W

Frontiers in physiology 20190306

Scopoletin is a promising acaricidal botanical natural compound against Tetranychus cinnabarinus, and its acaricidal mechanism maybe involve calcium overload according to our previous study. To seek potential candidate target genes of calcium overload induced by scopoletin in T. cinnabarinus, RNA-seq was utilized to detect changes in transcription levels. 24 and 48 h after treatment, 70 and 102 differentially expressed genes were obtained, respectively. Target genes included 3 sign ...[more]

PMID: 30894818

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Project description:IntroductionInvestigation into the action mechanisms of plant secondary metabolites against pests is a vital strategy for the development of novel promising biopesticides. Scoparone (isolated from Artemisia capillaris), a renewable plant-derived bioresource, displays potent acaricidal activities against mites, but its targets of action remain unclear.ObjectivesThis study aimed to systematically explore the potential molecular targets of scoparone against Tetranychus cinnabarinus and provide insights to guide the future application of scoparone as an agent for the management of agricultural mite pests worldwide.MethodsThe mechanism and potential targets of scoparone against mites were investigated using RNA-seq analysis; RNA interference (RNAi) assays; bioassays; and [Ca2+]i, pull-down and electrophysiological recording assays.ResultsRNA-seq analysis identified Ca2+ signalling pathway genes, specifically 5 calmodulin (CaM1-5) genes and 1 each of L-, T-, N-type voltage-gated Ca2+ channel (VGCC) genes, as candidate target genes for scoparone against mites. Furthermore, RNAi and electrophysiological data showed that the CaM1- and L-VGCC-mediated Ca2+ signalling pathways were activated by scoparone. Interestingly, by promoting the interaction between CaM1 and the IQ motif (a consensus CaM-binding domain of L-VGCC), CaM1 markedly enhanced the activating effect of scoparone on L-VGCC. Pull-down assays further demonstrated that CaM interacted with the IQ motif, triggering L-VGCC opening. Importantly, mutation of the IQ motif significantly weakened CaM1 binding and eliminated the CaM1-mediated enhancement of scoparone-induced L-VGCC activation, indicating that the effect of scoparone was dependent on the CaM1-IQ interaction.ConclusionThis study demonstrates, for the first time, that the acaricidal compound scoparone targets the interface between CaM1 and L-VGCC and activates the CaM-binding site, located in the IQ motif at the L-VGCC C-terminus. This work may contribute to the development of target-specific green acaricidal compounds based on L-VGCC.

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Acaricidal Mechanism of Scopoletin Against Tetranychus cinnabarinus.

Publications

Acaricidal Mechanism of Scopoletin Against <i>Tetranychus cinnabarinus</i>.

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