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The olfactory system of insects is crucial in modulating behaviors such as host seeking, mating, and oviposition. Odorant-binding proteins (OBPs) are involved in semiochemical recognition. OBPs recognize and bind odorants and transport them to odorant receptors located in olfactory neurons. Harmonia axyridis (Coleoptera: Coccinellidae) is a widely used predacious biological control agent for many agricultural and forestry pests. This study identified 19 OBPs in H. axyridis based on the antennal and whole-body transcriptomes of adults and obtained all the full-length open reading frames, including 11 ‘Classic’ OBPs, 7 ‘Minus-C’ OBPs and 1 ‘Plus-C’ OBP. They encoded 125 to 241 amino acid proteins with molecular weights ranging from 13.75 to 27.75 kDa and isoelectric points ranging from 4.15 to 8.80. Phylogenetic analyses were used to study the relationships between H. axyridis OBPs and OBPs from other species of Coleoptera. Quantitative real-time PCR (qPCR) analysis showed that HaxyOBP2, 3, 5, 8, 10, 12, 13, 14, and 15 were highly expressed in antennae of both adult females and males. Moreover, HaxyOBP2, 3, 5, 12, and 15 were more abundantly expressed in antennae than other body parts, while HaxyOBP13 and HaxyOBP14 were expressed predominantly, and at similar levels, in the head and antennae. The other OBP genes were highly expressed in non-olfactory tissues including the thorax, abdomen, legs, and wings. These results provide valuable information for further study of H. axyridis olfaction, which may ultimately enhance its use as a biocontrol agent.

Cyantraniliprole is a novel anthranilic diamide insecticide with significant efficacy against Bemisia tabaci, an important pest insect worldwide. In this study, we conducted reversion and selection work and genetic analysis, and determined cross-resistance spectrum and synergism of cyantraniliprole resistance based on the reported population, SX population, of B. tabaci collected from Shanxi Province, China. Compared with a susceptible strain (MED-S), SX population, the field-evolved cyantraniliprole-resistant population exhibited 26.4-fold higher resistance to cyantraniliprole. In SX, a sharp decline of cyantraniliprole resistance was shown in the absence of selection. Another tested strain, SX-R, was established from SX population after successive selection with cyantraniliprole and recently developed 138.4-fold high resistance to cyantraniliprole. SX-R had no cross-resistance to abamectin, imidacloprid, thiamethoxam, sulfoxaflor, or bifenthrin. Genetic analysis illustrated that cyantraniliprole resistance in SX-R was autosomally inherited and incompletely dominant. Additionally, piperonyl butoxide (PBO) significantly inhibited cyantraniliprole resistance in the SX-R strain. In conclusion, the selection of SX with cyantraniliprole led to high resistance to cyantraniliprole which is incompletely dominant and no cross-resistance to several common types of insecticides. Enhanced oxidative metabolism is possibly involved in the resistance of SX-R, yet target-site resistance could not be excluded.

Broflanilide, a novel meta-diamide insecticide, possesses moderate acute toxicity to zebrafish, with a 96-h median lethal concentration (96-LC

Bemisia tabaci is a threat to agriculture worldwide because of its potential to cause devastating damage to various crops. β-asarone is a bioactive pesticidal chemical originating from Acorus calamus (or “Sweet Flag”) plants, and it displays significant lethal effects against insect pests. In this study, we established a baseline of susceptibility to β-asarone from China and patterns of cross-resistance to other popular insecticides. We found that all the 12 field-collected B. tabaci populations exhibited high susceptibility to β-asarone, and there was no cross-resistance detected for other tested insecticides. We subsequently evaluated the sublethal effects of β-asarone on physiology and biochemistry via LC25 treatment (4.7 mg/L). LC25 of β-asarone resulted in prolonged developmental duration and decreased survival rates in B. tabaci nymphs, pseudopupae, and adults. Significant reductions in oviposition duration, fecundity, and hatchability were also observed. Additionally, the metabolic enzyme activity and expression profiles of selected cytochrome P450 monooxygenase (P450) genes following the LC25 treatment of β-asarone suggest that enhanced detoxification via P450s could be involved in the observed sublethal effects. These findings demonstrate the strong toxicity and significant sublethal effects of β-asarone on B. tabaci and suggest that the induced overexpression of P450 genes could be associated with the response to β-asarone.

The Bemisia tabaci (Gennadius) cryptic species complex comprises important insect pests that cause devastating damage to agricultural crops worldwide. In China, the B. tabaci Mediterranean (MED) (or biotype Q) species is threatening agricultural production all over the country as resistance to commonly used insecticides has increased. This situation highlights the need for alternative pest control measures. Cyantraniliprole, a novel anthranilic diamide insecticide, has been widely employed to control Hemipteran pests. To monitor the levels of resistance to cyantraniliprole in B. tabaci field populations in China, bioassays were conducted for 18 field samples from nine provinces over two years. Compared with median lethal concentration (LC50) for the MED susceptible strain, all field samples had significantly higher resistance to cyantraniliprole. Furthermore, resistance factors (RFs) increased significantly in samples from Shanxi (from 5.62 in 2015 to 25.81 in 2016), Hunan (3.30 in 2015 to 20.97 in 2016) and Hubei (from 9.81 in 2015 to 23.91 in 2016) provinces. This study indicates a considerable decrease in the efficacy of cyantraniliprole against B. tabaci and establishes a baseline of susceptibility that could serve as a reference for future monitoring and management of B. tabaci resistance to cyantraniliprole.

Harmonia axyridis (Coleoptera: Coccinellidae) is a polyphagous insect that is an important biological agent used to control agricultural and forestry pests. The role of functional genes in H. axyridis based on quantitative real-time PCR (qRT-PCR) is increasingly well understood to investigate biology, physiology, feeding behavior and the role of important genes in physiological processes. Quantitative real-time PCR (qRT-PCR) is a powerful and reliable technique to quantify gene expression. Using qRT-PCR, expression levels of target genes are determined based on the levels of internal reference genes; therefore, reference genes need to be stably expressed under specific experimental conditions. However, there have been no studies on the stability of reference genes used in H. axyridis. In this study, we systematically investigated expression profiles of nine candidate reference genes from H. axyridis, including β-actin (ACTIN); elongation factor 1 α (EF1A); ribosomal proteins L10, L18, L28, S13, and S15 (RPL10, RPL18, RPL28, RPS13 and RPS15); glyceraldehyde-3-phosphate dehydrogenase (GAPDH); and superoxide dismutase (SOD). Four analytical methods (geNorm, BestKeeper, NormFinder, and the ΔCt method) were used to evaluate the suitability of these genes as internal reference genes for three biotic factors (developmental stage, tissue, and sex) and two abiotic treatments (temperature and photoperiod). RefFinder, a comprehensive evaluation platform integrating the four analytical methods, was used to rank the overall stability of these reference genes. Among the nine candidate genes, different reference genes were identified as having the most stable expression across biotic and abiotic factors. Genes encoding ribosomal proteins typically had the most stable expression, though EF1A was the most stable across developmental stages and photoperiods. To validate the suitability of these reference genes, heat shock protein 90 (HSP90) was chosen as a target. Significant up-regulation in HSP90 expression level in response to both low and high temperature was observed when using the most suitable reference genes but not when using an arbitrarily selected reference gene. The reference genes identified in this study will provide the basis for future functional genomics research in H. axyridis and will also facilitate the establishment of a standardized qRT-PCR program for other related insects.