摘要: Lepidopteran insects produce cocoons with unique properties. The cocoons are made of silk produced in the larval tissue silk gland and our understanding of the silk genes is still very limited. Here, we investigated silk genes in the bagworm mothEumeta variegata, a species that has recently been found to produce extraordinarily strong and tough silk. Using short-read transcriptomic analysis, we identified a partial sequence of thefibroin heavy chaingene and its product was found to have a C-terminal structure that is conserved within nonsaturniid species. This is in accordance with the presence offibroin light chain/fibrohexameringenes and it is suggested that the bagworm moth is producing silk composed of fibroin ternary complex. This indicates that the fibroin structure has been evolutionarily conserved longer than previously thought. Other thanfibroins we identified candidates forsericingenes, expressed strongly in the middle region of the silk gland and encoding serine-rich proteins, and other silk genes, that are structurally conserved with other lepidopteran homologues. The bagworm moth is thus considered to be producing conventional lepidopteran type of silk. We further found a number of genes expressed in a specific region of the silk gland and some genes showed conserved expression withBombyx moricounterparts. This is the first study allowing comprehensive silk gene identification and expression analysis in the lepidopteran Psychidae family and should contribute to the understanding of silk gene evolution as well as to the development of novel types of silk.

摘要: Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), transmits the causal pathogen of huanglongbing and is a global pest of citrus. D. citri populations exhibit resistance to multiple insecticide modes of action in areas where these chemicals have been overused. We performed genome-wide transcriptional analysis for two field populations of D. citri (Wauchula and Lake Alfred, Florida, USA) that exhibit 1300-fold resistance to the neonicotinoid insecticide, thiamethoxam, and compared it to that of susceptible psyllids collected from the same area and without imposed selection. The Lake Alfred population responded to insecticide resistance by up-regulation of 240 genes and down-regulation of 148 others. The Wauchula population exhibited similar patterns to the Lake Alfred population with up-regulation of 253 genes and down-regulation of 115 others. Gene Ontology annotation associated with cellular processes, cell, and catalytic activity were assigned to differentially expressed genes (DEGs). The DEGs from Lake Alfred and Wauchula populations were mapped to Kyoto Encyclopedia of Gene and Genomes pathways and implicated enrichment of metabolic pathways, oxidative phosphorylation, extracellular matrix-receptor interaction, terpenoid backbone biosynthesis, and insect hormone biosynthesis in the resistant populations. Up-regulation of 60s ribosomal proteins, UDP-gluscoyltransferases, cytochrome c oxidases, and CYP and ABC transporters among thiamethoxam-resistant D. citri implicates a broad array of novel and conventionally understood resistance mechanisms.

摘要: RNA interference (RNAi) has emerged as a powerful tool for developing novel management strategies for controlling insect pests. The 28-spotted ladybeetle, Henosepilachna vigintioctopunctata is one of the most important pests attacking solanaceous plants in Asia. In this study, the potential of dietary RNAi to manage H. vigintioctopunctata was investigated using both in vitro synthesized and bacterially expressed double-stranded RNAs (dsRNAs) of HvvATPase A and HvvATPase E. The expression levels of HvvATPase A and HvvATPase E were higher in Malpighian tubules than in other tissue types. The silencing of HvvATPase A and HvvATPase E led to significant mortality in H. vigintioctopunctata larvae. In addition, the ingestion of HvvATPase A and HvvATPase E significantly deterred feeding behavior and subsequently arrested the development of H. vigintioctopunctata. Notably, the bacterially expressed dsRNAs consistently caused higher mortality in larvae and adults. Finally, the nontarget effects of the dsRNAs of H. vigintioctopunctata on the predatory ladybeetle Propylaea japonica were evaluated. P. japonica 1st instar larvae were administered vATPase A and vATPase E dsRNAs from H. vigintioctopunctata and P. japonica under the worst-case scenario, in which dsGFP served as negative control. There were significant effects of dsHvvATPase A on P. japonica at the transcriptional level but not at the organismal level, whereas dsHvvATPase E did not effect P. japonica at either the transcriptional or the organismal level. Collectively, the results of the study suggest that HvvATPase A and HvvATPase E can act as novel molecular targets for the control of H. vigintioctopunctata.

摘要: Granulocytes and plasmatocytes play important roles in clearing foreign objects in insects, but it is difficult to distinguish between them in immune reactions. Based on the hemocyte cell line SYSU-OfHe-C established at our lab, two cell sublines, SYSU-OfHe-C Granulocyte (Gr cells) and SYSU-OfHe-C Plasmatocyte (Pl cells), which possess the morphological characteristics of granulocytes and plasmatocytes, respectively, were established. Gr and Pl cells showed different behaviors in immune reactions, such as spreading, phagocytosis and encapsulation. Pl cells were easier to spread, but Gr cells tended to undergo aggregation, indicating that they may take different strategies to clear foreign objects. These results also suggested that granulocytes and plasmatocytes may express some different proteins. By comparing the gene expression in cells from the two sublines, 1662 differentially expressed genes were identified, and 13 out of 30 transmembrane proteins highly expressed in Pl cells (six) or Gr cells (seven) were further screened and confirmed by reverse-transcription polymerase chain reaction (PCR). Finally, three transmembrane genes specifically expressed in Pl cells and two transmembrane genes specifically expressed in Gr cells were screened out based on their expressions in immune reactions by quantitative PCR analysis. These genes may potentially be used as molecular markers to distinguish between granulocytes and plasmatocytes inOstrinia furnacalis, and further to clarify the functions of immune hemocytes in cellular immune reaction such as encapsulation and so on.

摘要: Validamycin has been widely used as a specific competitive inhibitor of trehalase. In our previous research, validamycin significantly inhibited trehalase activity and chitin synthesis in Diaphorina citri, resulting in abnormal phenotypes. However, the mechanism of validamycin's action on D. citri remains unclear. Here, using a comparative transcriptome analysis, 464 differentially expressed genes (DEGs) in D. citri were identified after validamycin treatment. A Gene Ontology enrichment analysis revealed that these DEGs were mainly involved in small molecule process, structural molecule activity and transition metal ion binding. DEGs involved in chitin metabolism, cuticle synthesis and insecticide detoxification were validated by reverse transcription quantitative polymerase chain reaction. The RNA interference of D. citri chitinase-like protein ENO3 and D. citri cuticle protein 7 genes significantly affected D. citri molting. Moreover, the recombinant chitinase-like protein ENO3 exhibited a chitin-binding property, and an antimicrobial activity against Bacillus subtilis. This study provides a first insight into the molecular changes in D. citri after exposure to validamycin and identifies two effective RNA interference targets for D. citri control.