Genome wide gene-expression analysis of facultative reproductive diapause in the two-spotted spider mite Tetranychus urticae
ABSTRACT: Investigating essential physiological processes in diapausing mites by analyzing genome wide gene expression changes using custom-built microarray. We investigated the molecular biology of facultative reproductive diapause in the chelicerate Tetranychus urticae (Acari: Tetranychidae) by analyzing genome-wide gene expression differences in diapausing and non diapausing T. urticae, using an Agilent custom-built two color gene expression microarray. Analysis of this dataset showed that a remarkable number, 11% of the total number of predicted T. urticae genes, were differentially expressed. Gene Ontology analysis revealed that many metabolic pathways were affected in diapausing females. Genes related to digestion and detoxification, cryo-protection, carotenoid synthesis and the organization of the cytoskeleton were profoundly influenced by the state of diapause. We also further confirmed the importance of horizontally transferred carotenoid synthesis genes in diapause and different color morphs of T. urticae. We made one comparison: diapausing mites (DIA) vs non-diapausing mites (NON-DIA), in 4 replicates. Both types of mites belonged to the T. urticae LS-VL strain. In this strain, approximately 30% of mites enter diapause under the experimental conditions applied. Hence, we were able to sample RNA of mites with similar genetic background that were reared under identical environmental conditions. The labeled cRNA samples were pooled and hybdrized to a custom Sureprint genome wide G3 Gene Expression 8x60K microarray. Data was normalized by Agilent Feature Extraction software (using protocol GE2_107_SEP09). Genespring software (Agilent technologies) was used for the statistical analysis of the data.
Project description:This SuperSeries is composed of the following subset Series: GSE31525: Spider mite preliminary feeding experiment with mites reared on bean and two Arabidopsis thaliana accessions GSE31527: Developmental stage-specific gene expression in the two-spotted spider mite (Tetranychus urticae) GSE32005: Developmental stage-specific small RNA composition in the two-spotted spider mite (Tetranychus urticae) GSE32009: Transcriptional responses of the two-spotted spider mite (Tetranychus urticae) after transfer to different plant hosts Refer to individual Series
Project description:Cyflumetofen is a novel acaricide on the international market with an unknown mode of action. Under laboratory conditions, we selected for high levels of cyflumetofen resistance in a susceptible mite strain and performed genome-wide gene-expression analysis. Differential expression between the resistant and susceptible strain was identified to study on the molecular level how mites develop resistance to this novel acaricide. Our results show that in T. urticae, the selection for cyflumetofen resistance resulted in a differential expression in only a limited number of genes. This transcriptomic study provides an unbiased look how cyflumetofen resistance triggers selection on the transcriptional level in T. urticae. 4 replicates for one comparison; per replicate 150 mites were pooled. Mites of the derived cyflumetofen resistant spider mite (cy5-labelled) were directly compared to mites of the ancestral susceptible strain (cy3-labelled).
Project description:We sequenced messenger RNA from mixed stages of the two-spotted spider mite (Tetranychus urticae) reared on bean (Phaseolus vulgaris cv California Red Kidney; the laboratory host plant for mites) and two Arabidopsis thaliana accessions which were considered to either be susceptible (Kondara) or resistant (Bla-2) to mite feeding. This pilot experiment was conducted to assess gene expression differences of mites grown on sensitive versus resistant Arabidopsis accessions, as well as differences in mites feeding on different host species. The expression data was used for gene model validation of genes predicted by EuGene in the spider mite genome and to assess gene expression levels. Examination of gene expression of spider mites reared on beans and two Arabidopsis accessions (Kondara and Bla-2).
Project description:While pathogen-induced immunity is comparatively well characterized, far less is known about plant defense responses to arthropod herbivores. To date, most molecular-genetic studies of plant-arthropod interactions have focused on insects. However, plant-feeding (phytophagous) mites are also pests of diverse plants, and mites induce different patterns of damage to plant tissues than do well-studied insects (e.g., Lepidopteran larvae or aphids). The two-spotted spider mite, Tetranychus urticae, is among the most significant mite pests in agriculture. T. urticae is an extreme generalist that has been documented on a staggering number of plant hosts (more than 1,100), and is renowned for the rapid evolution of pesticide resistance. To understand reciprocal interactions between T. urticae and a plant host at the molecular level, we examined mite herbivory using Arabidopsis thaliana. Despite differences in feeding guilds, we found that transcriptional responses of A. thaliana to mite herbivory generally resembled those observed for insect herbivores. In particular, defense to mites was mediated by jasmonic acid (JA) biosynthesis and signaling. Further, indole glucosinolates dramatically increased mite mortality and development times. Variation in both basal and activated levels of these defense pathways might also explain differences in mite damage and feeding success between A. thaliana accessions. On the herbivore side, a diverse set of genes associated with detoxification of xenobiotics was induced upon exposure to increasing levels of in planta indole glucosinolates. Our findings provide molecular insights into the nature of, and response to, herbivory for a representative of a major class of arthropod herbivores. We used microarray to assess global gene expresion in Arabidopsis thaliana upon Tetranychus urticae attack in two A. thaliana accessions: Bla-2, resistant to spider mite herbivory and Kon, susceptible to spider mite herbivory. 3 week old Arabidopsis thaliana plants were subjected to Tetranychus urticae attack through application of 10 mites for various periods of time (timecourse scenario) or hundreds of mites for 1 hour (feeding site scenario).
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.
Project description:Our goal was to investigate the transcriptional response of T. urticae to different Arabidopsis plants with varying levels of indole glucosinolates. Bean is the ancestral host plant for the T. urticae strain used in this study. Adult female spider mites were transferred to the three Arabidopsis lines (Col-0, qKo, atr1D) or bean, and after 24 hours, the gene expression levels were analyzed. 12 samples were investigated; 4 biological replicates for three comparisons: QKO (qKo vs. Col-0), ATR1D (atr1D vs. Col-0), and COL-0 (Col-0 vs. bean).
Project description:The two-spotted spider mite, Tetranychus urticae, is one of the most significant mite pests in agriculture that can feed on more than 1,100 plant hosts, including model plants Arabidopsis thaliana and tomato, Solanum lycopersicum. Here, we described tomato transcriptional responses to spider mite feeding and compared them to Arabidopsis in order to determine conserved and divergent responses to this pest. 2,133 differentially expressed genes (DEGs) were detected at 1, 3, 6, 12 or 24 hours post spider mite infestation (hpi) relative to non-infested control plants. Based on Biological Process Gene Ontology annotations, improved in the course of our analysis, DEGs were grouped in 60 significantly enriched gene sets that highlighted perception of the spider mite attack (1 hpi), metabolic reprogramming (3-6 hpi), and establishment and maintenance of the defense responses (6-24 hpi). We used microarray to assess global gene expression in Solanum lycopersicum cv. Heinz 1706 upon Tetranychus urticae attack. 1 month old tomato plants were subjected to Tetranychus urticae attack through application of 100 adult mites on a terminal leaflet of leaf 3 for various periods of time (timecourse scenario) or hundreds of mites for 1 hour (feeding site scenario).
Project description:The most common ladybird beetle, Coccinella septempunctata L., is an excellent predator of crop pests such as aphids and white flies, and it shows a wide range of adaptability, a large appetite and a high reproductive ability. In this study, we collected female adults in three different states, i.e., non-diapause, diapause and diapause termination, for transcriptome sequencing. The experimental insects consisted of three different states as follows: Non-diapause female insects were reared at 24±1°C, with a RH of 70±10% and a 16:8 h light: dark (L: D) photoperiod and collected after their first oviposition. Female adults in diapause were reared at 18±1°C at an RH of 70±10% and a 10:14-h (L:D) photoperiod. The experimental diapause insects were collected after 30 days. Diapause-terminated adults were transferred to another climatic cabinet with the 30-day diapause insects and reared under the same conditions as the non-diapause insects. After their first oviposition, the female insects were collected and stored at -80°C. Three biological replicates per treatment (non-diapause, diapause, diapause-terminated) were sequenced using Illumina HiSeq 2500.
Project description:250 adult T. urticae females from the London strain (grown on acyanogenic P. vulgaris cv. Prelude bean plants) were transferred to cyanogenic P. lunatus cv. 8078 bean plants. Thirty-five generations after the host transfer, total RNA was extracted from mites growing on both bean species (London and London-CYANO strain) and used in in a genome-wide gene expression microarray (Sureprint G3 microarray, Agilent) experiment to assess significantly differentially expressed genes (FC ≥ 2 and FDR-corrected p-value < 0.05) between mites grown on P. vulgaris (cv. Prelude) bean plants (London strain) and mites grown for 35 generations on P. lunatus (cv. 8078) bean plants (London-CYANO strain). 4 replicates for one comparison: mites of the London strain grown on P. lunatus for 35 generations (London-CYANO) compared to mites of the London strain grown on P. vulgaris bean plants (London)