Project description:Heortia vitessoides (Moore) is the most destructive defoliating pests in Aquilaria sinensis (Loureiro) Sprenger forests in Southern China. The complete sequences of mitochondria is reported: a circular molecule of 15,516?bp in size included 40.13% for A, 40.79% for T, 11.23% for C and 7.86% for G. There are 60 genes including 3 species with 12 protein-coding genes, 2 different species ribosomal RNA genes (S and L rRNA species), 46 transfer RNA genes (20 RNA species). H. vitessoides (Moore) and other 19 species belonging to lepidopteran were phylogenetic and analyzed by MEGA 6.06 with neighbor-joining methods. The mtDNA of H. vitessoides (Moore) were clustered in lepidopteran superfamilies.
Project description:Juvenile hormone diol kinase (JHDK) is a critical enzyme involved in juvenile hormone degradation in insects. In this study, HvJHDK in the Heortia vitessoides Moore (Lepidoptera: Crambidae) transcriptional library was cloned. Stage-specific expression patterns of HvJHDK, HvJHEH, and HvJHE as well as juvenile hormone titers were determined. The three tested enzymes participated in juvenile hormone degradation. Moreover, juvenile hormone titers peaked after larval-larval molts, consistent with a role for juvenile hormone in inhibition of metamorphosis. HvJHDK was subsequently suppressed using RNA interference (RNAi) to reveal its functions. Different concentrations of dsJHDK elicited the optimal interference efficiency at different life stages of H. vitessoides. Suppression of HvJHDK decreased HvJHDK content and increased the juvenile hormone titer, thereby resulting in reduced triglyceride content, sharply declined survival rate, clearly lethal phenotypes, and extended larval growth. Moreover, suppression of HvJHDK upregulated HvJHEH and HvJHE expression levels, suggesting that there is feedback regulation in the juvenile hormone metabolic pathway. Taken together, our findings provide molecular references for the selection of novel insecticidal targets.
Project description:To elucidate the role of glutathione S-transferases (GSTs) in Heortia vitessoides Moore (Lepidoptera: Crambidae), one of the most destructive defoliating pests in Aquilaria sinensis (Lour.) Gilg (Thymelaeaceae) forests, 16 GST cDNAs were identified in the transcriptome of adult H. vitessoides. All cDNAs included a complete open reading frame and were designated HvGSTd1-HvGSTu2. A phylogenetic analysis showed that the 16 HvGSTs were classified into seven different cytosolic classes; three in delta, two in epsilon, three in omega, three in sigma, one in theta, two in zeta, and two in unclassified. The expression patterns of these HvGSTs in various larval and adult tissues, following exposure to half the lethal concentrations (LC50s) of chlorantraniliprole and beta-cypermethrin, were determined using real-time quantitative polymerase chain reaction (RT-qPCR). The expression levels of the 16 HvGSTs were found to differ among various larval and adult tissues. Furthermore, the RT-qPCR confirmed that the transcription levels of nine (HvGSTd1, HvGSTd3, HvGSTe2, HvGSTe3, HvGSTo3, HvGSTs1, HvGSTs3, HvGSTu1, and HvGSTu2) and six (HvGSTd1, HvGSTd3, HvGSTe2, HvGSTo2, HvGSTs1, and HvGSTu1) HvGST genes were significantly higher in the fourth-instar larvae following exposure to the insecticides chlorantraniliprole and beta-cypermethrin, respectively. These genes are potential candidates involved in the detoxification of these two insecticides. Further studies utilizing the RNA interference approach are required to enhance our understanding of the functions of these genes in this forest pest.
Project description:Autophagy is a highly conserved and regulated process in eukaryotic cells and remodels cytoplasm, recovers essential nutrients, and disposes of unwanted cytoplasmic components. Autophagy-related gene (ATG) 8, identified in Heortia vitessoides Moore, which is an oligophagous pest of Aquilaria sinensis (Lour.), was characterized (HvATG8). Multiple sequence alignment showed that HvATG8 possesses highly conserved domain structures. Stage- and tissue-specific expressions indicated that HvATG8 is highly expressed in prepupal, pupal, and adult stages and in the midgut of larvae and abdomen of adults. Lack of function of HvATG8 by RNA interference resulted in a significant decrease in survival rate and an increase in abnormal or nonviable phenotypes in H. vitessoides. Transition rate from larval to pupal stages was 33.0% and from pupal to adult stages was 15.0% after injection. Reduction of ATG8 expression reduced survival of H. vitessoides. Therefore, HvATG8 possibly plays a key role in normal growth stage of H. vitessoides. HvATG8 suppression downregulates HvATG3 expression, suggesting that the two genes are interconnected. Further, HvATG8 expression increased by 20-hydroxyecdysone treatment, starvation, and extreme temperature exposure. Starvation also altered expression of other ATGs in H. vitessoide. This study may be used to guide research on molecular mechanisms of autophagy in insects.