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In insects, the gustatory system has a critical function not only in selecting food and feeding behaviours but also in growth and metabolism. Gustatory receptors play an irreplaceable role in insect gustatory signalling. Trichogramma chilonis is an effective biocontrol agent against agricultural insect pests. However, the molecular mechanism of gustation in T. chilonis remains elusive. In this study, we found that T. chilonis adults had a preference for D-fructose and that D-fructose contributed to prolong longevity and improve fecundity. Then, We also isolated the full-length cDNA encoding candidate gustatory receptor (TchiGR43a) based on the transcriptome data of T. chilonis, and observed that the candidate gustatory receptor gene was expressed from the larval to adult stages. The expression levels of TchiGR43a were similar between female and male. A Xenopus oocyte expression system and two-electrode voltage-clamp recording further verified the function analysis of TchiGR43a. Electrophysiological results showed that TchiGR43a was exclusively tuned to D-fructose. By the studies of behaviour, molecular biology and electrophysiology in T. chilonis, our results lay a basic fundation of further study on the molecular mechanisms of gustatory reception and provide theoretical basis for the nutritional requirement of T. chilonis in biocontrol.
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???displayArticle.pmcLink???PMC6583964 ???displayArticle.link???PLoS One
Fig 1. Behavioral preference to D-fructose of T. chilonis.A: a, T. chilonis fed nothing; b, T. chilonis fed both fructose and water; c, T. chilonis fed water; d, T. chilonis fed sugar. B: Relative sensitivity of D-fructose was determined by two-choice preference tests. PI values for D-fructose are shown at the following concentrations: 0.010, 0.025, 0.050, 0.100 and 0.300 M every concentration was tested with 50â60 adults. Error bars indicate SEMs from the analysis of three replications (P < 0.05).
Fig 2. Longevity and fecundity of female T. chilonis when supplied with D-fructose, water and nothing.(A) Longevity, error bars indicate SEMs from the analysis of 30 replications (P < 0.05).; (B) Fecundity, error bars indicate SEMs from the analysis of 60 replications (P < 0.05).
Fig 3. Phylogenetic analysis of putative gustatory receptors of T. chilonis.The tree was constructed in MEGA6.0 using the neighbor-joining method. TchiGR43a from T. chilonis are labelled with red, GRs from D. melanogaster (Diptera) are labelled with blue, GRs from B. mori (Lepidoptera) are labelled with purple, and GRs from other Hymenoptera insects (T. pretiosum, N. vitripennis, Apis mellifera, C. floridanum, Cephus cinctus, Orussus abietinus, Pseudomyrmex gracilis and Athalia rosae) are labelled with green.
Fig 4. Expression patterns of TchiGR43a in T. chilonis.(A) Relative expression levels of TchiGR43a in different developmental stages of T. chilonis by qRT-PCR analysis. Larval stage: d2, prepupal stage: d3-d4, pupal stage: d5-d8. (B) Relative expression levels of TchiGR43a between male and female adult T. chilonis by qRT-PCR analysis. Error bars indicate SEMs from the analysis of three replications (P < 0.05).
Fig 5. Two-electrode voltage-clamp recordings of Xenopus oocytes expressing TchiGR43a isolated in the present study.(A) Inward current responses of the oocytes expressing TchiGR43a in response to 0.100 M solution of the 11 sugars. (B) Xenopus oocytes with no injection. (C) Xenopus oocytes injected with ddH2O. (D) Inward current responses of the oocytes expressing TchiGR43a in response to 0.100 M solution of D-fructose and myo-inositol (mean ± SEM (n = 5)). (E) The oocytes expressing TchiGR43a stimulated with a range of D-fructose concentrations. (F) Dose-response curve of the oocytes expressing TchiGR43a to D-fructose. EC50 = 0.023 M. Bars indicate SEM (n = 6).
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