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MicroRNAs (miRNAs), a class of non-coding single-strand RNA molecules encoded by endogenous genes, are about 21-24 nucleotides long and are involved in the post-transcriptional regulation of gene expression in plants and animals. Generally, the types and quantities of miRNAs in the different tissues of an organism are diverse, and these divergences may be related to their specific functions. Here we have identified 296 known miRNAs and 46 novel miRNAs in the antennae of the parasitoid wasp Microplitis mediator by high-throughput sequencing. Thirty-three miRNAs were predicted to target olfactory-associated genes, including odorant binding proteins (OBPs), chemosensory proteins, odorant receptors (ORs), ionotropic receptors (IRs) and gustatory receptors. Among these, 17 miRNAs were significantly highly expressed in the antennae, four miRNAs were highly expressed both in the antennae and head or wings, while the remaining 12 miRNAs were mainly expressed in the head, thorax, abdomen, legs and wings. Notably, miR-9a-5p and miR-2525-3p were highly expressed in male antennae, whereas miR-1000-5p and novel-miR-13 were enriched in female antennae. The 17 miRNAs highly expressed in antennae are likely to be associated with olfaction, and were predicted to target one OBP (targeted by miR-3751-3p), one IR (targeted by miR-7-5p) and 14 ORs (targeted by 15 miRNAs including miR-6-3p, miR-9a-5p, miR-9b-5p, miR-29-5p, miR-71-5p, miR-275-3p, miR-1000-5p, miR-1000-3p, miR-2525-3p, miR-6012-3p, miR-9719-3p, novel-miR-10, novel-miR-13, novel-miR-14 and novel-miR-28). These candidate olfactory-associated miRNAs are all likely to be involved in chemoreception through the regulation of chemosensory gene expression in the antennae of M. mediator.

BACKGROUND: The Apolygus lucorum is one of the most destructive insect pests in China with a wide range of host plants. Interaction of A. lucorum with surrounding environment heavily relies on chemical communication. Deorphanization of receptors involved in odors detection elevates our understanding of the olfactory system of this pest and may help to develop a chemical ecology‐based control strategy. RESULTS: AlucOR80, an odorant receptor (OR) in A. lucorum was newly cloned. Gene expression analysis showed that this receptor was mainly expressed in the antennae and head of both sexes but with a male bias. The Xenopus oocytes heterologous expression system coupled with the two‐electrode voltage‐clamp (TEVC) recording revealed that AlucOR80 was tuned to 21 selected compounds. Furthermore, electroantennogram (EAG) tests confirmed that all 21 ligands of AlucOR80 were electrophysiologically active in antennae of both sexes. Behavioral trials in a three‐cage olfactometer indicated that 16 compounds were behaviorally active, amongst which, 12 components were attractants and four components were repellents for adults of both sexes. Butyl butyrate and Dimethyl disulfide (DMDS) were the strongest attractive and repellant compounds, respectively. Importantly, we found the repellency of 1, 8‐Cineole, S‐(−)‐cis‐Verbenol and (1S)–(1)‐beta‐Pinene against adults of A. lucorum. CONCLUSION: Although AlucOR80 is a general OR, may play important role in the olfactory perception of A. lucorum. Screening of AlucOR80 ligands by behavioral assay provided valuable insights by which olfactory‐based management approaches could be developed by utilizing the behaviorally active components as attractants or repellents. © 2019 Society of Chemical Industry

Sensory neuron membrane proteins (SNMPs), homologs of the human fatty acid transport protein CD36 family, are observed to play a significant role in chemoreception, especially in detecting sex pheromone in Drosophila and some lepidopteran species. In the current study, two full‐length SNMP transcripts, MmedSNMP1 and MmedSNMP2, were identified in the parasitoid Microplitis mediator (Hymenoptera: Braconidae). Quantitative real‐time polymerase chain reaction analysis showed that the expression of MmedSNMP1 was significantly higher in antennae than in other tissues of both sexes. In addition, the MmedSNMP1 transcript was increased dramatically in newly emerged adults and there were no significant differences between adults with or without mating and parasitic experiences. However, compared with MmedSNMP1, the expression of MmedSNMP2 was widely found in various tissues, significantly increased at half‐pigmented pupae stage and remained at a relatively constant level during the following developmental stages. It was found that MmedSNMP1 contained eight exons and seven introns, which was highly conserved compared with other insect species. In situ hybridization assay demonstrated that MmedSNMP1 transcript was distributed widely in antennal flagella. Among selected chemosensory genes (odorant binding protein, odorant receptor, and ionotropic receptor genes), MmedSNMP1 only partially overlapped with MmedORco in olfactory sensory neurons of antennae. Subsequent immunolocalization results further indicated that MmedSNMP1 was mainly expressed in sensilla placodea of antennae and possibly involved in perceiving plant volatiles and sex pheromones. These findings lay a foundation for further investigating the roles of SNMPs in the chemosensation of parasitoids.

Laboratory bioassays were conducted to determine the effect of food source on the survival of Tribolium castaneum Herbst, Oryzaephilus surinamensis L. and Rhyzopertha dominica F., after exposure to five diatomaceous earth (DE) formulations: Protect-It ® , Insecto ® , Perma-Guard TM , Dryacide ® and SilicoSec ® . Adults of these species were exposed to DEs at the rate of 0.5 mg/cm 2 for 1 day on filter paper inside plastic Petri dishes. After exposure, the initial mortality was counted and live individuals of the three species were held for a week in glass vials containing 50 mg wheat flour, rice and whole wheat, respectively. In the second experiment, after 1 day exposure to DEs, beetles were transferred to Petri dishes without food and held for a week to determine if the presence of food source would decrease the mortality of beetles. Experiments were carried out at 27 o C and 55% RH in the dark. The initial mortality in both of the experiments reached 100% for the three species exposed to Protect-It ® and in the case of R. dominica and O. surinamensis exposed to Dryacide ® . In contrast, low level of mortality (< 10%) was observed for T. castaneum exposed to Perma-Guard TM and Insecto ® . The mortality after the post-treatment period on food was decreased for the three species exposed to Perma-Guard TM and in the case of T. castaneum and R. dominica exposed to Insecto ® and SilicoSec ® . Adults of O. surinamensis were the most susceptible followed by R. dominica and 100% adult mortality was obtained, whereas T. castaneum were the least susceptible beetles to DEs. Protect-It ® and Dryacide ® were the most efficient DE formulations and can be used effectively in a stored grain integrated pest management program.