Project description:RNA-seq was employed to analyze the transcriptome changes in T. cinnabarinus treated with scopoletin or solvent. A total of 104, 783, 164 clean sequence reads were generated from the sequencing with more than 90% of the reads were successfully mapped to the reference sequence. The RNA-seq identified 70 and 102 differentially expressed genes between scopoletin and solvent treated mites at 24 h and 48 h post treatment, respectively. GO enrichment analysis showed that differentially expressed genes from 24 h post treatment were categorized into 31 GO functional groups and differentially expressed genes from 48 h post treatment were categorized into 29 GO functional groups. The cellular process under the category of biological process was dominant throughout the GO classification at both time points. Overall our results revealed the global transcriptional changes in T. cinnabarinus upon scopoletin treatment and will enable the further identification of the targets of scopoletin on mite.
Project description:RNA-seq was employed to analyze the transcriptome changes in T. cinnabarinus treated with curcumin or solvent. A total of 105, 706, 297 clean sequence reads were generated from the sequencing with more than 90% of the reads were successfully mapped to the reference sequence. The RNA-seq identified 111 and 96 differentially expressed genes between curcumin and solvent treated mites at 24 h and 48 h post treatment, respectively. GO enrichment analysis showed that differentially expressed genes from 24 h post treatment were categorized into 35 GO functional groups and differentially expressed genes from 48 h post treatment were categorized into 25 GO functional groups. The cellular process under the category of biological process was dominant throughout the GO classification at both time points. Finally, we screened 23 differentially expressed genes that are functionally identical or similar to the targets of common insecticide/acaricides or associated with mite detoxification and metabolism. Overall our results revealed the global transcriptional changes in T. cinnabarinus upon curcumin treatment and will enable the further identification of the targets of curcumin on mite.
Project description:We generated 38-bp Illumina reads from single messenger RNA libraries from three diverse developmental stages of the two-spotted spider mite to capture small RNA diversity across development. Adult, nymphal+larvae and embryonic stages were separated using sieves of various pore sizes, and mites of various developmental stages were carefully selected for small RNA library preparation. Samples were a mix of males and females to capture male and female patterns of small RNA composition and were reared on beans (Phaseolus vulgaris cv California Red Kidney). Small RNA reads were used for miRNA prediction, piRNA discovery, and for quantitation of small RNA-generating loci (i.e. expression across development). Examination of small RNA from spider mites of adult, embryonic and pooled larval/nymphal developmental stages.
Project description:We generated 38-bp Illumina reads from single messenger RNA libraries from three diverse developmental stages of the two-spotted spider mite to capture small RNA diversity across development. Adult, nymphal+larvae and embryonic stages were separated using sieves of various pore sizes, and mites of various developmental stages were carefully selected for small RNA library preparation. Samples were a mix of males and females to capture male and female patterns of small RNA composition and were reared on beans (Phaseolus vulgaris cv California Red Kidney). Small RNA reads were used for miRNA prediction, piRNA discovery, and for quantitation of small RNA-generating loci (i.e. expression across development).
Project description:We generated 77-bp Illumina reads from single messenger RNA libraries from four diverse developmental stages of the two-spotted spider mite to maximally capture the complement of transcribed sequences across development. Adult, nymphal, larvae and embryonic stages were separated using sieves of various pore sizes, and mites of various developmental stages were carefully selected for transcriptome library preparation. Samples were a mix of males and females to capture male and female patterns of transcription, and were reared on beans (Phaseolus vulgaris cv California Red Kidney). The RNA-Seq data was used for validation of gene models predicted by EuGene, and to study patterns of gene expression across development. Gene expression for spider mites from adult, nymph, larvae and embryonic developmental stages was examined (technical replicates were generated).
Project description: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.
Project description:We generated 38-bp Illumina reads from messenger RNA libraries from mites transferred from their preferred laboratory host, bean (Phaseolus vulgaris cv. California Red Kidney), to one of three hosts: bean, Arabidopsis thaliana (Bla-2 accession) and the tomato (Solanum lycopersicum; genotype Heinz 1706). Larvae were carefully collected from bean plants and transferred to the treatment plant. Mites were reared on these plants for ~24 hours, after which mites were collected for mRNA library preparation. Samples were a mix of males and females. The goal of the study was to identify genes that may underlie the ability of mites to be herbivores on different host plants.
Project description:Purpose: The present study provides the firstly large-scale characterization of miRNAs in Tetranychus cinnabarinus and the comparison between fenpropathrin resistant and susceptible strains gives a clue on study how miRNA involving in fenpropathrin resistance Methods: Using Illumina sequencing to identify the differentially expressed miRNAs between the fenpropathrin resistant and susceptible strains of Tetranychus cinnabarinus Results: 12 miRNAs that were expressed significantly differently were identified between thethe fenpropathrin resistant and susceptible strains of Tetranychus cinnabarinus Total RNA obtained from 500 female adults aged 3-5 days post emergence from fenpropathrin resistant and susceptible strains of Tetranychus cinnabarinus,two biological repeats for each strain Raw sequencing read data were submitted to SRA: SRP067789
Project description:The two-spotted spider mite Tetranychus urticae is an extreme polyphaguous crop pest. Next to an increased detoxification potential of plant secondary metabolites, it has recently been shown that spider mites manipulate plant defences. Salivary constituents are proposed to play an important role during the interaction with its many hosts. The proteomic composition of saliva delivered into artificial diet by spider mites adapted to various hosts - bean, soy, maize, tomato -was determined using Orbitrap mass spectrometry. Over 200 different proteins were identified, many of unknown function and in numerous cases belonging to multi-membered gene families. A selection of these putative salivary proteins was validated using whole-mount in situ hybridizations and expression was shown to be localized in the anterior and dorsal podocephalic glands of the spider mite. Host-plant dependent expression was evident from the proteomics dataset and was further studied in detail by micro-array based genome wide gene expression profiling of mites maintained on the host plants under study. Previously obtained gene-expression datasets were further used to get more insight in the expression profile over different life stages and physiological states. To conclude, for the first time the T. urticae salivary proteome repertoire was characterized using a custom feeding hemisphere-based enrichment technique. This knowledge will assist in unraveling the molecular interactions between phytophagous mites and their host plants. This may ultimately facilitate the development of mite-resistant crops.