Project description:The early embryo of the spider Parasteatoda tepidariorum initially shows spherical symmetry in morphology. Following a symmetry breaking event, it forms a radially symmetric germ disc in the hemisphere of the egg. In this work, we conducted RNA-sequencing of cells isolated from small different regions of radially symmetric forming/formed germ discs, aming to identify candidate genes whose transcripts are locally expressed in early spider embryos.
Project description:Spider silk proteins are synthesized in the silk-producing glands, where the spidroins are produced, stored and processed into a solid fiber from a crystalline liquid solution. Despite great interest in the spider silk properties, that make this material suitable for biomedical and biotechnological applications, the mechanism of formation and spinning of the silk fibers has not been fully elucidated; and no combination of proteomic and transcriptomic study has been carried out so far in the spider silk-producing glands. Nephila clavipes is an attractive orb-web spider to investigate the spinning process of silk production, given the properties of strength, elasticity and biocompatibility of their silk fibers. Thus, considering that the combination of proteomic and transcriptomic analysis may reveal an extensive repertoire of novel proteins involved in the silk spinning process, and in order to facilitate and enable proteomics in this non-model organism, the current study aims to construct a high quality reference mRNA-derived protein database that could be used to identify tissue specific expression patterns in spider silk glands. Next-generation sequencing has offered a powerful and cost-efficient technique for the generation of transcriptomic datasets in non-model species using diverse platforms such as the Illumina HiSeq, Roche 454, Pacific Biosystems, and Applied Biosystems SOLiD; In the current study, the Illumina HiSeq 2000 platform will be used to generate a N. clavipes spider silk glands transcriptome-based protein database. The transcriptome data generated in this study will provide a comprehensive and valuable genomic resource for future research of the group of spider silk-producing glands, in order to improve our understanding of the overall mechanism of action involved in production, secretion, storage, transport, protection and conformational changes of spidroins during the spinning process, and prey capture; and the results may be relevant for scientists in material Science, biology, biochemistry, and environmental scientists.
Project description:The common house spider Parasteatoda tepidariorum is a chelicerate model organism for studying developmental mechanisms and their evolution in arthropods. In contrast to the well-studied model insect, Drosophila melanogaster, embryos of the spider undergo patterning in a cellular environment from early stages (at least after the number of the nuclei increase to 16). Use of spider embryos provide new opportunities to understand the evolution of developmental mechanisms underlying arthropod body plans. This analysis aims to generate genome-scale, developmental profiles of gene expression in embryos of the spider P. tepidariorum, which facilitate a wide range of studies using this spider species.
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:Restriction site Associated DNA (RAD) tags are a genome-wide representation of every site of a particular restriction enzyme by short DNA tags. Most organisms segregate large numbers of DNA sequence polymorphisms that disrupt restriction sites, which allow RAD tags to serve as genetic markers spread at a high-density throughout the genome. Here, we demonstrate the applicability of RAD markers for both individual and bulk-segregant genotyping. First, we show that these markers can be identified and typed on pre-existing microarray formats. Second, we present a method that uses RAD marker DNA to rapidly produce a low-cost microarray genotyping resource that can be used to efficiently identify and type thousands of RAD markers. We demonstrate the utility of the former approach by using a tiling path array for the fruit fly to map a recombination breakpoint, and the latter approach by creating and utilizing an enriched RAD marker array for the threespine stickleback. The high number of RAD markers enabled localization of a previously identified region, as well as a second novel region also associated with the lateral plate phenotype. Taken together, our results demonstrate that RAD markers, and the method to develop a RAD marker microarray resource, allow high-throughput, high-resolution genotyping in both model and non-model systems. Keywords: microarray genotyping
Project description:In a cellular field of the early spider embryo, Hedgehog signaling operates to specify a “fuzzy” French-flag-like pattern along the primary axis. We applied single-cell and single-nucleus RNA sequencing to the early spider embryo. We confirmed that these techniques successfully detected three cell population corresponding to germ layers and some known cell types. We showed that the data had sufficient information for reconstruction of a correct global polarity of the presumptive ectoderm.
Project description:Callobius koreanus (C.koreanus) is a wandering spider and a member of the Amaurobiidae family, infraorder Araneae. RNA-sequencing was performend for venom gland tissue and whole body except venom gland.
Project description:Agelena koreana is indigenous spider in South Korea that lives on piles of trees building webs. RNA-sequencing was performed for venom gland tissue and whole body except venom gland.
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