Project description:The molecular mechanism involved in BmNPV resistance was investigated using a genome wide microarray in the midgut tissues of BmNPV infected resistant (Sarupat) and susceptible (CSR-2) Indian silkworm races. In the resistant race, 735 genes were upregulated and 589 genes were down regulated at 12 hours of BmNPV post infection. Similarly in case of susceptible race, 2183 genes were up regulated and 2115 genes were down regulated. Among the differentially expressed genes, nine upregulated and eight down regulated genes were validated using real time qPCR analysis. In Sarupat, the significantly upregulated genes are vacuolar protein sorting associated gene, X fin like protein and Carboxy peptidase E like protein in BmNPV infected midguts, whereas the prominent down regulated genes are glutamate receptor ionotropic kainite 2-like, BTB/POZ domain and transferrin. In the case of CSR-2, the considerably upregulated genes are Peptidoglycan recognition protein S6 precursor and rapamycin while the conspicuous down regulated genes are facilitated trehalose transporter and zinc transporter ZIP1-like gene. Our results provided a vital insights of Bombyx mori in reference with its molecular mechanism in immune response against the BmNPV invasion. Organism : Bombyx mori , Agilent Custom Silkworm Gene Expression 4x44k Array (AMADID: 066047) designed by Genotypic Technology Private Limited.
Project description:We identified genes regulated by parasitization of the silkworm Bombyx mori by three tachinid parasitoid species, Exorista japonica, Drino inconspicuoides and Pales pavida, using oligonucleotide microarrays. The numbers of genes and their intensity of expression varied with the species of parasitoid, within silkworm hemocytes and fat body. Bombyx mori hemocyte, silkgland and fat body samples parasitizated by Exorista japonica, Drino inconspicuoides and Pales pavida were prepared. Gene expression was compared in these two groups: control and parasitized.
Project description:Uric acid (UA) is the final product of purine metabolism and plays an important role as a physiological antioxidant. In recent years, several different groups have reported a correlation between decreased UA in Parkinson’s disease (PD) and clinical progression and stage of PD. However, little is known about the molecular mechanisms of decreased UA under oxidative stress. We used our systematic functional annotation pipeline for silkworm genes to identify a novel UA metabolic pathway regulator under oxidative stress in a UA metabolism mutant silkworm Bombyx mori model. Gene expression was measured in 3day of fifth instar larvae of abnormal uric acid synthesis Bombyx mori mutant of op.
Project description:Background: MicroRNAs (miRNAs) are expressed by a wide range of eukaryotic organisms, and function in diverse biological processes. Numerous miRNAs have been identified in Bombyx mori, but the temporal expression profiles of miRNAs corresponding to each stage transition over the entire life cycle of the silkworm remain to be established. To obtain a comprehensive overview of the correlation between miRNA expression and stage transitions, we performed a whole-life test and subsequent stage-by-stage examinations on nearly one hundred miRNAs in the silkworm. Results: Our results show that miRNAs display a wide variety of expression profiles over the whole life of the silkworm, including continuous expression from embryo to adult (bmo-miR-184), up-regulation over the entire life cycle (bmo-let-7 and miR-100), down-regulation over the entire life cycle (miR-124), expression associated with embryogenesis (miR-29 and miR-92), up-regulation from early 3rd instar to pupa (miR-275), and complementary pulses in expression between miR-34b and miR-275. Stage-by-stage examinations revealed further expression patterns, such as emergence at specific time-points during embryogenesis and up-regulation of miRNA groups in late embryos (miR-1 and bantam), expression associated with stage transition between instar and molt larval stages (miR-34b), expression associated with silk gland growth and spinning activity (miR-274), continuous high expression from the spinning larval to pupal and adult stages (miR-252 and miR-31a), a coordinate expression trough in day 3 pupae of both sexes (miR-10b and miR-281), up-regulation in pupal metamorphosis of both sexes (miR-29b), and down-regulation in pupal metamorphosis of both sexes (miR-275). Conclusions: We present the full-scale expression profiles of miRNAs throughout the life cycle of Bombyx mori. The whole-life expression profile was further investigated via stage-by-stage analysis. Our data provide an important resource for more detailed functional analysis of miRNAs in this animal. In the study presented here, we performed a whole-life test and subsequent stage-by-stage examinations on nearly one hundred miRNAs in the silkworm, leading to a comprehensive overview of the correlation between miRNA expression and stage transitions. In all, 59 unique developmental timepoints were inspected in this study.
Project description:Background: The growth and development of the posterior silk gland and the biosynthesis of the silk core protein at the fifth larval instar stage of Bombyx mori are of paramount importance for silk production. Results: Here, aided by next-generation sequencing and microarry assay, we profile 1,229 microRNAs (miRNAs), including 728 novel miRNAs and 110 miRNA/miRNA* duplexes, from the posterior silk gland at the fifth larval instar. Target gene prediction yields 14,222 unique target genes from 1,195 miRNAs. Functional categorization classifies the genes into complex pathways that include both cellular and metabolic processes, especially protein synthesis and processing. Conclusion: The enrichment of target genes in the ribosome-related pathway indicates that miRNAs may directly regulate translation. Our findings pave a way for further functional elucidation of these miRNAs in silk production. Sequencing 10 total RNA samples from the posterior silk gland of different strains and developmental stage using Illumina Solexa technology. Four strains of silkworm (Q, B, QB and BQ) with different two development stages (stage 1: fourth instar molting to day 2 of fifth instar; stage 2: fifth instar day 3 to day 8 before spinning, according to our previous genes expression cluster analysis), and two strains (R1 and J1) from entire period (stage 1 + stage 2).
Project description:Microsporidia have attracted much attention because they infect a variety of species ranging from protists to mammals, including immunocompromised patients with AIDS or cancer. Aside from the study on Nosema ceranae, few works have focused on elucidating the mechanism in host response to microsporidia infection. Nosema bombycis is a pathogen of silkworm pébrine that causes great economic losses to the silkworm industry. Detailed understanding of the host (Bombyx mori) response to infection by N. bombycis is helpful for prevention of this disease. The 23 K silkworm genome array was used to investigate host responses (i.e., Bombyx mori) occurring at 2, 4, 6 and 8 d post-infection by Nosema bombycis.We focused on elucidating the mechanism of the host response to microsporidia infection, especially for the investigation of host immune response . The third instar molted silkworm larvae were in oral infected by Nosema bombycis. In order to known the silkworm host response to Nosema bombycis infection at different time points, samples of infected larvae (i.e., the treatment set) and uninfected larvae (i.e., the control set) were collected at 2, 4, 6 and 8 dpi for RNA extraction and array hybridization. The obtained data were usd to investigate on the interplay of the genome-wide expression profile of hosts.
Project description:To reveal the variations in transcriptome responses of hemocytes of B. mori after the parasitic infection. Overall design: Organism : Bombyx mori , Agilent Custom Silkworm Gene Expression 4x44k Array (AMADID: 066047) designed by Genotypic Technology Private Limited.
Project description:Using 4 replicate males and 4 replicate females this experiment examined dosage compensation and sex-biased gene expression. Briefly we performed a de novo assembly of the Manduca sexta transcriptome using all sequenced libraries, quantified genes expression, identified the physical locations of genes through orthology to the moth Bombyx mori and examine expression differences between autosomal and Z-linked genes between males and females. Further, we examined sex-biased gene expression using the replicated data.
Project description:Molecular genetic studies of Bombyx mori have led to profound advances in understanding the regulation of development. Bombyx mori brain, as a main endocrine organ, plays important regulatory roles in various biological processes. The microarray technology will allow the genome-wide analysis of gene expression patterns in silkworm brains. We reported microarray-based gene expression profiles in silkworm brains at four stages including V7, P1, P2 and P3. A total of 4,550 genes were transcribed in at least one selected stage. Of these, clustering algorithms separated the expressed genes into stable expressed genes and variable expressed genes. The results of the gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) analysis of stable expressed genes showed that the ribosomal and oxidative phosphorylation pathways were principal pathways. Secondly, four clusters of genes with significantly different expression patterns were observed in the 1,175 variable expressed genes. Thirdly, thirty-two neuropeptide hormones genes, nine neuropeptide-like precursor genes, and 117 cuticular protein genes were expressed in selected developmental stages. The present study deﬁned major characteristics of the transcriptional profiles in the brains of Bombyx mori at the specific development stages. Our data will provide abundant information that will be useful in future research. Transcription profiling experiments, 4 developmental stages (samples) were analyzed. Dual-channel experiments, with test samples labeled by Cy5 and common reference samples labeled by Cy3. Common reference sample was used for data normalization. One Biological replicate. No dye-swaps.
Project description:In the silkworm, Bombyx mori, juvenile hormone (JH) and 20-hydroxyecdysone (20E) levels are high during the final larval molt (4M) but both absent during the feeding stage of 5th instar (5F), while JH level is low and 20E level is high during the prepupal stage (PP). Fat body is the important organs in insect, we want to find out differentially expressed genes which are respectively regulated by the two hormones. Total RNA from 4th molting,5th feeding and prepupa stages Bombyx fat body were used to generate target cDNA, and then hybridized to 48k Bombyx genome Array Genechips, representing about 23000 characterized genes