Project description:Juvenile hormone (JH) is produced and secreted by corpora allata (CA) during larval stages. JH biosynthesis from acetyl-CoA includes 13 enzymatic steps. Genes for 10 of those enzymes have been identified from Bombyx mori. It was also reported that they are basically expressed selectively in CA. Genes for the other 3 enzymes were recently reported from other insects, i.e., farnesyl diphosphate pyrophosphatase (FPPP) from Drosophila and farnesol dehydrogenase (FolD) and farnesal dehydrogenase (FalD) from Aedes. In order to reveal the expression profiles of these genes in Bombyx CA and to find other candidate genes involved in JH biosynthesis, we performed a transcriptomic analysis in corpora cardiaca-CA (CC-CA) complexes using a custom-made DNA microarray on which sequences from CC-CA EST libraries and genome-widely predicted genes are loaded. CC-CA complexes vigorously expressed JH-biosynthetic genes identified in Bombyx as well as genes involved in methyl-group metabolism and ribosomal protein genes; this is suitable for JH production. Bombyx counterparts of Drosophila FPPP and Aedes FolD genes, however, showed quite low level of expression. On the other hand, some other genes probably encoding oxidoreductases were expressed highly and selectively in CC-CA complex. Their products might function as FolD and/or FalD in Bombyx.
Project description:Differentially expressed microRNAs (miRNAs) were detected to explore the molecular mechanisms of diapause termination. The total small RNA (sRNA) of diapause-destined silkworm eggs and HCl-treated eggs was extracted and then sequenced using HiSeq/MiSeq high-throughput method. 44 novel miRNAs were discovered. Compared to those in the diapause-destined eggs, 61 miRNAs showed significant changes in the acid-treated eggs, with 23 being up-regulated and 38 being down-regulated. The potential target genes of differentially expressed miRNAs were predicted by miRanda. Gene Ontology and KEGG pathway enrichment analysis of these potential target genes revealed that they were mainly located within cells and organelles, involved in cellular and metabolic processes, and participated in protein production, processing and transportation. Two differentially expressed genes, Bombyx mori SDH (BmSDH) and Bmo-miR-2761-3p, were further analyzed with qRT-PCR. BmSDH was significantly up-regulated in the HCl-treated eggs, while Bmo-miR-2761-3p was down-regulated. These results suggested that these two genes were well coordinated in silkworm eggs. Dual luciferase reporter (DLR) assay demonstrated that Bmo-miR-2761-3p inhibited the expression of BmSDH.