Dataset Information

Functional Analysis of Esterase TCE2 Gene from Tetranychus cinnabarinus (Boisduval) involved in Acaricide Resistance.

ABSTRACT: The carmine spider mite, Tetranychus cinnabarinus is an important pest of crops and vegetables worldwide, and it has the ability to develop resistance against acaricides rapidly. Our previous study identified an esterase gene (designated TCE2) over-expressed in resistant mites. To investigate this gene's function in resistance, the expression levels of TCE2 in susceptible, abamectin-, fenpropathrin-, and cyflumetofen-resistant strains were knocked down (65.02%, 63.14%, 57.82%, and 63.99%, respectively) via RNA interference. The bioassay data showed that the resistant levels to three acaricides were significantly decreased after the down-regulation of TCE2, indicating a correlation between the expression of TCE2 and the acaricide-resistance in T. cinnabarinus. TCE2 gene was then re-engineered for heterologous expression in Escherichia coli. The recombinant TCE2 exhibited ?-naphthyl acetate activity (483.3?±?71.8?nmol/mg pro. min(-1)), and the activity of this enzyme could be inhibited by abamectin, fenpropathrin, and cyflumetofen, respectively. HPLC and GC results showed that 10??g of the recombinant TCE2 could effectively decompose 21.23% fenpropathrin and 49.70% cyflumetofen within 2?hours. This is the first report of a successful heterologous expression of an esterase gene from mites. This study provides direct evidence that TCE2 is a functional gene involved in acaricide resistance in T. cinnabarinus.

SUBMITTER: Shi L

PROVIDER: S-EPMC4698665 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature

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Functional Analysis of Esterase TCE2 Gene from Tetranychus cinnabarinus (Boisduval) involved in Acaricide Resistance.

Shi Li L Wei Peng P Wang Xiangzun X Shen Guangmao G Zhang Jiao J Xiao Wei W Xu Zhifeng Z Xu Qiang Q He Lin L

Scientific reports 20160104

The carmine spider mite, Tetranychus cinnabarinus is an important pest of crops and vegetables worldwide, and it has the ability to develop resistance against acaricides rapidly. Our previous study identified an esterase gene (designated TCE2) over-expressed in resistant mites. To investigate this gene's function in resistance, the expression levels of TCE2 in susceptible, abamectin-, fenpropathrin-, and cyflumetofen-resistant strains were knocked down (65.02%, 63.14%, 57.82%, and 63.99%, respec ...[more]

PMID: 26725309

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Project description:ObjectiveTetranychus cinnabarinus (Boisduval) is an agricultural mite pest threatens crops throughout the world, causing serious economic loses. Exploring the effects of acaricides on predatory mites is crucial for the combination of biological and chemical control of T. cinnabarinus. Neoseiulus californicus (McGregor) is one of the principal natural enemies of T. cinnabarinus, which can be applied in protected agriculture. In this study, the effects of sublethal concentrations of a new acaricide, SYP-9625 on two mite species, and the effects of the application concentration on predatory mite, N. californicus were assessed. The aim of the present study was to evaluate the effect of SYP-9625 on life parameters and predation capacity of N. californicus based on the concentration-response bioassay of T. cinnabarinus to explor the application of the new acaricide with natural enemy N. californicus.MethodAll of the experiments were conducted under laboratory conditions [25 ± 1°C, 16: 8 h (L: D) and 75 ± 5% RH]. The sublethal concentrations LC10 (0.375μg/mL) and the LC30 (0.841μg/mL) against T. cinnabarinus and the application concentration (100μg/mL) against N. californicus were used to evaluate the effects of SYP-9625 on population parameters of N. californicus based on an age-stage, two-sex life table and its predation capacity by functional response.Resultcinnabarinus females treated with LC30 exhibited significantly reduced net reproductive rates (R0 = 11.02) in their offspring compared with females treated with LC10 (R0 = 14.96) and untreated females (R0 = 32.74). However, the intrinsic rate of increase (rm) and the finite rate of increase (λ) of N. californicus indicated that the application concentration of SYP-9625 had no significant negative effect on N. californicus eggs (rm = 0.277, λ = 1.319) compared to the control (rm = 0.292, λ = 1.338). Additionally, most population parameters of N. californicus showed a dose-dependent manner with the increase of the concentration of SYP-9625 against T. cinnabarinus. SYP-9625 also stimulated the control efficiency of N. californicus against immobile stages including eggs and larvae.ConclusionThis study demonstrated that sublethal concentrations of SYP-9625 can inhibit the population growth of T. cinnabarinus. In addition, the sublethal concentrations and the application concentration showed no effect on the population growth of N. californicus. These two advantages described above showed great commercial potential of this new acaricide based on population parameters of the two mite species and predation capacity of the predatory mite under laboratory conditions.

Project description:The carmine spider mite, Tetranychus cinnabarinus (Boisduval), is one of the most important acarine pest species. At present, its control remains primarily dependent on using various chemical insecticides/acaricides in agricultural crops worldwide. To clarify the mechanism whereby T. cinnabarinus responds to insecticide exposure, we identified the chitin synthase 1 gene (TcCHS1) and then explored the gene expression levels of TcCHS1 at different developmental stages of T. cinnabarinus. We also investigated the effects of sublethal concentrations of diflubenzuron on the toxicities and survivals of T. cinnabarinus eggs and larvae as well as TcCHS1 expression levels. The full-length cDNA sequence contains an open reading frame (ORF) of 4881 nucleotides that encoded for a 1474 amino acid residues protein. The predicted TcCHS1 protein had a molecular mass of 168.35 kDa and an isoelectric point of 6.26, and its amino acid sequence contained all the signature motifs (EDR, QRRRW and TWGTR) of chitin synthases. The results of phylogenetic analyses demonstrated that the putative CHS1 amino acid sequence of T. cinnabarinus revealed high similarities with chitin synthases in other insects and mites. Additionally, at the molecular level, transcriptional analysis by real-time quantitative PCR in different developmental stages of T. cinnabarinus revealed that TcCHS1 mRNA was expressed in all stages, and highest in eggs and female adults, but lowest in deutonymphs. Furthermore, the results of toxicity bioassays indicated that diflubenzuron treatment resulted in high mortality rates in eggs and larvae of T. cinnabarinus. The mRNA expression levels of TcCHS1 from the eggs and larvae of T. cinnabarinus were up-regulated in response to sublethal concentrations of diflubenzuron exposures. Together, all these results demonstrate that diflubenzuron has ovicidal and larvicidal effects and TcCHS1 may play an important role in the growth and development of T. cinnabarinus and may disrupt the chitin biosynthesis, thereby controlling T. cinnabarinus populations.

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.

Dataset Information

Functional Analysis of Esterase TCE2 Gene from Tetranychus cinnabarinus (Boisduval) involved in Acaricide Resistance.

Publications

Functional Analysis of Esterase TCE2 Gene from Tetranychus cinnabarinus (Boisduval) involved in Acaricide Resistance.

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