Conopomorpha sinensis Bradley is the dominant borer pest of Litchi chinesis and Euphoria longan. Current management of C. sinensis relies upon insecticide application to adult moths. In addition to the direct mortality induced by insecticides, a sublethal dose of insecticides also affects growth, survival, and reproduction in the exposed insects. Vitellogenin (Vg) and vitellogenin receptor (VgR) are normally identified as essential reproduction-related proteins in insects. In this study, we characterized these two genes from C. sinensis, and investigated their differential responses to sublethal concentrations of insecticide. Cloned CsVg and CsVgR consist of 5391 and 5424-bp open reading frames, which encode proteins of 1796 and 1807 amino acid residues, respectively. The CsVg protein contains the typical vitellogenin, DUF1943 and VWFD domains as other reported lepidopteran Vgs. The CsVgR was characterized as a typical low density lipoprotein receptor with two highly conserved LBD and EGF precursor domains, one hydrophobic transmembrane domain, one cytoplasmic domain, and 13 putative N-glycosylation sites. We next assessed the sublethal effect of four major insecticides on egg-laying in C. sinensis. The toxicity against C. sinensis varied among the insecticides tested, with LC50 values ranging from 0.23 ppm for chlorpyrifos to 20.00 ppm for β-cypermethrin, among which emamectin benzoate (EB) showed a significant negative impact on egg-laying, survival rate, ovarian development, and mating rate of C. sinensis at LC30 doses. Further investigation showed that the transcriptional level of CsVg and CsVgR were impaired in different way at 24, 48, and 72 h after EB exposure, and this result was in agreement with the diminished egg-laying of C. sinensis in the sublethal concentration EB-treated group. A repressed transcription level of CsVgR was observed at 48 h after treatment, suggesting that EB elicits a delayed response in the abundance of CsVgR. These results established different roles of CsVg and CsVgR in response to the sublethal effect of insecticides. CsVg might be a better parameter than CsVgR for assessing the effect of sublethal insecticides on reproduction in C. sinensis.

BackgroundThe peach fruit moth, Carposina sasakii Matsumura (Lepidoptera: Carposinidae), poses a serious threat to a variety of fruits and causes significant economic loss owing to difficulties in its prevention and control. The olfactory sense is generally acknowledged to be a novel target for pest control. However, a systematic study of the olfactory genes expressed in C. sasakii has not been reported yet. Here, we reported the antennal transcriptome of C. sasakii using high-throughput sequencing and annotated the main chemosensory multi-gene families.ResultsIn the chemosensory gene families, 29 odorant-binding proteins, 13 chemosensory proteins, 1 sensory neuron membrane protein, 52 odorant receptors, 8 ionotropic receptors and 11 gustatory receptors were annotated in the C. sasakii antennal transcriptome. The number of olfactory genes obtained in our transcriptome was consistent with that identified in other lepidopteran insects, confirming that we basically accomplished the annotation of the chemosensory genes of C. sasakii in the adult antennal transcriptome. All sequences were annotated and analyzed by BLAST (basic local alignment search tool), and some chemosensory genes with specific functions were named according to the BLAST results and phylogenetic trees. Based on the expression profile in the transcriptome and phylogenetic analysis, differentially expressed genes (DEGs) were analyzed in both male and female adults. Finally, fluorescence quantitative real-time PCR was used to identify the male-specific or female-specific chemosensory genes that were putatively related to odor detection and recognition. Moreover, expression levels of OR33 and PBP2 were significantly higher in males than in females, indicating that these genes may interact with sex pheromones. We found some conserved antennal IRs and GRs involved in detecting sugar compounds (GR2, GR5, GR6, GR8) and carbon dioxide (GR1), which were also identified based on phylogenetic analysis.ConclusionsThere are 114 putative chemosensory proteins expressed in C. sasakii identified in this study. The identification of these proteins will make the molecular mechanism of odor recognition accessible.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-4900-x) contains supplementary material, which is available to authorized users.

The purpose of this study was to evaluate the sublethal effects of the beta-cypermethrin on calling behavior and fecundity of a major fruit-boring pest of apple, Carposina sasakii Matsumura. The mating rate, fertility (total number of eggs laid per female), and adult longevity of adults were remarkably decreased as compared with that in control when the adults were exposed to 10% lethal concentrations (LC10) of beta-cypermethrin (LC10♀ × LC10♂), and the age-specific survival rate (lx) was also negatively affected by sublethal beta-cypermethrin especially for the LC10♀× LC10♂ mating combination. However, the age-specific fecundity (mx) was stimulated particularly in the combination of CK♀× LC10♂. Furthermore, the oviposition period was prolonged and the number of eggs was significantly increased for combinations of CK♀× LC10♂ and LC10♀× CK♂. In the mating experiments, males in control or LC10-beta-cypermethrin treatments preferred to mate with females in control. It might be because of lower calling rate of female survivors treated with sublethal beta-cypermethrin. Our data indicate that treatment of beta-cypermethrin had a sublethal effect on the development and production of C. sasakii, and their mating behavior changes in surviving adults that may contribute to assortative mating.

Conopomorpha sinensis is the dominant borer pest of Litchi chinensis (litchi) and Euphoria longan (longan) in China. Control of C. sinensis is difficult because of its cryptic life habit; thus, an effective ovicide could be beneficial. The larvicidal effects of diflubenzuron (DFB) have been documented in many insect pest species. Therefore, DFB might be a useful ovicide to control C. sinensis. However, the detailed mode of action of DFB interference with insect molting and egg hatching is unclear. Thus, we studied alterations in expression of all genes potentially affected by DFB treatment using a transcriptome approach in 2-d-old C. sinensis eggs. Clean reads were assembled to generate 203 455 unigenes and 440 558 transcripts. A total of 4625 differently expressed genes, which included 2670 up-regulated and 1955 down-regulated unigenes, were identified. Chitin binding and chitin metabolic processes were among the most significant enriched pathways according to Gene Ontology analyses. Most of the genes that encode enzymes involved in the chitin biosynthesis pathway were unaffected, whereas genes that presumably encode cuticle proteins were up-regulated. Furthermore, altered expression patterns of 10 genes involved in the chitin biosynthesis pathway of C. sinensis embryos were observed in response to DFB treatment at different time points by quantitative reverse transcription polymerase chain reaction. We also observed abnormal development; there was reduced chitin content and modulated chitin distribution of newly hatched larvae, and altered egg hatching. Our findings illustrate an ovicidal effect of DFB on C. sinensis, and reveal more molecular consequences of DFB treatment on insects.

Conopomorpha sinensis Bradley (Lepidoptera: Gracilariidae) is the dominant insect pest of litchi (chinensis Sonn.) and longan (Euphoria longan Lour.) fruit trees. Management of this pest species is a challenging task due to its cryptic borer behavior. Controlling C. sinensis at the egg stage is the best alternative strategy to chemical control of C. sinensis adults. However, thorough studies regarding the indirect and sublethal effects of chemicals on the different developmental stages of C. sinensis are insufficient. In this study, the effect of some insecticides was evaluated on C. sinensis eggs. The ovicidal activity of chlorbenzuron, abamectin, chlorantraniliprole, and λ-cyhalothrin was confirmed by morphological observation of the defects in C. sinensis eggs. Moreover, we characterized four essential ecdysone receptor proteins in insects [i.e., two isoform ecdysone receptors (EcR: CsEcRA. CsEcRB) and two isoform ultraspiracle proteins (USP: CsUSP1, CsUSP2)] from C. sinensis eggs. The CsEcRA, CsEcRB, CsUSP1, and CsUSP2 genes consisted of 1521-, 1614-, 1410-, and 1236-bp open reading frames which encoded proteins of 506, 527, 469, and 413 amino acid residues, respectively. Furthermore, the embryonic differential responses of CsEcRs, CsUSPs, and vitellogenin receptor (VgR: CsVgR) to insecticides were evaluated by qRT-PCR. Among the five tested genes, CsVgR and CsUSP1 were the most sensitive to all the tested insecticides, with fold change of the expression diminished by 4.27-8.70 times compared with untreated control insects. The data suggests that these insecticidal compounds regulate the expression of these specific proteins, which might eventually lead to reduced viability of C. sinensis eggs. We present here the first data providing molecular elucidation of ecdysone receptor genes and their differential responses to insecticides in C. sinensis eggs. Together with our previous report of insecticide sublethal effects on two reproduction-related genes in C. sinensis adults, CsVgR and CsUSP1

Conopomorpha sinensis Bradley is the dominant borer pest of litchi and longan in the Asian-pacific area. Reduction or interference of reproduction and mating of adult moths is one of the most used strategies to control C. sinensis. Insect reproduction is a critical biological process closely related to endocrine control. Conopomorpha sinensis genome and transcriptome information is limited, hampering both our understanding of the molecular mechanisms underlying hormone activity and reproduction and the development of control strategies for this borer pest. To explore the sex differences in gene expression profiles influencing these biological processes, de novo transcriptomes were assembled from female and male adult C. sinensis specimens. This analysis yielded 184,422 unigenes with an average length of 903 bp and 405,961 transcripts after sequencing and assembly. About 45.06, 22.41, 19.53, 34.05, 35.82, 36.42, and 19.85% of the unigenes had significant matches in seven public databases. Subsequently, gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis revealed comprehensive information about the function of each gene and identified enriched categories and pathways that were associated with the 2,890 female-biased genes and 2,964 male-biased genes. In addition, we identified some important unigenes related to hormone activity and reproduction among the sex-differentially expressed genes (DEGs), including unigenes coding for ecdysone-induced protein 78C, juvenile hormone (JH)-regulated gene fatty acyl-CoA reductase, vitellogenin, etc. Our findings provide a more comprehensive portrait of the sex differences involved in the relationship of two important physiological features—hormone activity and reproduction in C. sinensis and members of the family Gracillariidae.

Whether and how insecticide exposure will affect the biological control efficacy of predatory arthropods is critical in insecticide toxicology research but largely unexplored. In the current study, reduced biocontrol efficacy was observed in a predatory stink bugEocanthecona furcellataafter insecticide application in the field. Thus, we constructed a comparative transcriptome analysis and identified a total of 4364 upregulated and 1043 down regulated differentially expressed genes following the sublethal exposure of λ-cyhalothrin. The reduced juvenile hormone (JH) titer and increased trehalose content were observed. The predation capacity and theoretical maximum predation of predators were decreased by 31.08 and 48.90% in response to λ-cyhalothrin, respectively. Furthermore, JH supplementation after λ-cyhalothrin treatment could significantly stimulate trehalase and detoxification enzyme activities, as well as restore the predatory ability of E. furcellata. Our results help to understand the toxicological mechanism of predatory stink bug species in responding to insecticides, benefit predators' ecological services, and optimize the insecticide selection.

BACKGROUND: Chemosensory proteins (CSPs) play a vital role in the response to environmental stimuli in insects. However, the involvement of insect CSPs in the stress response to night-time environmental changes has not been examined.RESULTS: In the current study, four TiCSP genes were first cloned from Thalassodes immissaria by transcriptome and RACE-PCR techniques. TiCSPs had typical characteristics of insect CSPs, including a highly conserved four-cysteine motif and olfactoryspecific protein D (OS-D) or OS-D superfamily domains. TiCSP1-4 were clustered classified within different clades in a phylogenetic analysis and were differentially expressed at all developmental stages. Under night-time artificial light stress, the expression levels of TiCSP1 in males were significantly decreased at 24 h, and those of TiCSP2 were decreased in both adult sexes at 48 h. In a molecular docking analysis, TiCSPs showed relatively higher binding affinities with sex pheromone components than with host plant volatile molecules.CONCLUSION: Taking the reduced expression levels of TiCSPs and binding affinities into account, TiCSP1 and TiCSP2 are involved in the stress response processes of T. immissaria under light treatment. Our study supplies basic data for the evaluation of the effects of light interference control technologyan emerging physical control measure on nontarget pests of lychee orchards.