Project description:The striped flea beetle, Phyllotreta striolata, is one of the most destructive pests of Cruciferae crops worldwide. RNA interference (RNAi) is a promising alternative strategy for pest biological control, which overcomes the weakness of synthetic insecticides, such as pest resistance, food safety problems and toxicity to non-target insects. The homolog of Spt16/FACT, dre4 plays a critical role in the process of gene transcription, DNA repair, and DNA replication; however, the effects of dre4 silencing in P. striolata remain elusive. In this study, we cloned and characterized the full-length dre4 from P. striolata and silenced Psdre4 through microinjection and oral delivery; it was found that the silencing of dre4 contributed to the high mortality of P. striolata in both bioassays. Moreover, 1166 differentially regulated genes were identified after Psdre4 interference by RNA-seq analysis, which might have been responsible for the lethality. The GO analysis indicated that the differentially regulated genes were classified into three GO functional categories, including biological process, cellular component, and molecular function. The KEGG analysis revealed that these differentially regulated genes are related to apoptosis, autophagy, steroid hormone biosynthesis, cytochrome P450 and other signaling pathways. Our results suggest that Psdre4 is a fatal RNAi target and has significant potential for the development of RNA pesticides for P. striolata management.
Project description:The ability of a specialized herbivore to overcome the chemical defense of a particular plant taxon not only makes it accessible as a food source but may also provide metabolites to be exploited for communication or chemical defense. Phyllotreta flea beetles are adapted to crucifer plants (Brassicales) that are defended by the glucosinolate-myrosinase system, the so-called "mustard-oil bomb." Tissue damage caused by insect feeding brings glucosinolates into contact with the plant enzyme myrosinase, which hydrolyzes them to form toxic compounds, such as isothiocyanates. However, we previously observed that Phyllotreta striolata beetles themselves produce volatile glucosinolate hydrolysis products. Here, we show that P. striolata adults selectively accumulate glucosinolates from their food plants to up to 1.75% of their body weight and express their own myrosinase. By combining proteomics and transcriptomics, a gene responsible for myrosinase activity in P. striolata was identified. The major substrates of the heterologously expressed myrosinase were aliphatic glucosinolates, which were hydrolyzed with at least fourfold higher efficiency than aromatic and indolic glucosinolates, and β-O-glucosides. The identified beetle myrosinase belongs to the glycoside hydrolase family 1 and has up to 76% sequence similarity to other β-glucosidases. Phylogenetic analyses suggest species-specific diversification of this gene family in insects and an independent evolution of the beetle myrosinase from other insect β-glucosidases.
Project description:Olfactory transduction is a process by which olfactory sensory neurons (OSNs) transform odor information into neuronal electrical signals. This process begins with the binding of odor molecules to receptor proteins on olfactory receptor neuron (ORN) dendrites. The major molecular components involved in olfaction include odorant-binding proteins (OBPs), chemosensory proteins (CSPs), odorant receptors (ORs), gustatory receptors (GRs), ionotropic receptors (IRs), sensory neuron membrane proteins (SNMPs) and odorant-degrading enzymes (ODEs). More importantly, as potential molecular targets, chemosensory proteins are used to identify novel attractants or repellants for environmental-friendly pest management. In this study we analyzed the transcriptome of the flea beetle, Phyllotreta striolata (Coleoptera, Chrysomelidae), a serious pest of Brassicaceae crops, to better understand the molecular mechanisms of olfactory recognition in this pest. The analysis of transcriptomes from the antennae and terminal abdomens of specimens of both sexes identified transcripts from several key molecular components of chemoreception including 73 ORs, 36 GRs, 49 IRs, 2 SNMPs, 32 OBPs, 8 CSPs, and four candidate odorant degrading enzymes (ODEs): 143 cytochrome P450s (CYPs), 68 esterases (ESTs), 27 glutathione S-transferases (GSTs) and 8 UDP-glycosyltransferases (UGTs). Bioinformatic analyses indicated that a large number of chemosensory genes were up-regulated in the antennae. This was consistent with a potential role in olfaction. To validate the differential abundance analyses, the expression of 19 genes encoding various ORs, CSPs, and OBPs was assessed via qRT-PCR between non-chemosensory tissue and antennae. Consistent with the bioinformatic analyses, transcripts for all of the genes in the qRT-PCR subset were elevated in antennae. These findings provide the first insights into the molecular basis of chemoreception in the striped flea beetle.