| Olfaction plays a crucial role in the reproduction and survival of insect populations,so the identification of olfactory-related genes will facilitate the study of insect chemical communication systems.Mythmna separate is an important agricultural pest in China,which can feed 16 families and 104 plants.In this study,male and female antennae transcriptome of Mythimna separata?Walker?were sequenced using RNA-seq technology.A total of 71.71,66.42,63.37,60.56,57.64 and 67.21 M Clean reads were obtained,with each data volume greater than 8 G.Values of Q30 are greater than 90%,and GC content is around 45%.Three replicates of the same sample were assembled to obtain 326,538?average length 608 bp?transcripts and 132,454?average length 1021 bp?Unigenes,of which 200-500 bp genes were the most.The obtained genes were aligned in 7 large databases,among which the number of genes annotated in the NR database was the largest,accounting for 28.40% of the total number of genes,followed by the GO database?24.03%?and the Pfam database?23.9%?.GO function annotation found that binding?17487/54.92%?and catalytic activity?12190/38.29%?annotated the most numerous genes in the molecular function,which is consistent with the functional characteristics of insect antennae.We identified 57 ORs genes,22 IRs genes and 28 GRs genes through the annotation and keyword search from the male and female antennae transcriptome data of M.separata,and analyzed the spatiotemporal expression of 5 PRs genes.The results showed that the 5 PRs genes were specifically expressed in male and female antennae,and the expressions of Msep PR1,Msep PR2,Msep PR4 and Msep PR5 in the antennae of female were significantly higher than male,while the expressions of Msep PR3 was significantly higher expressed in the antennae of male than female.The expression level of Msep PR2 and Msep PR4 before mating was significantly higher than that after mating.The expression levels of PRs gene in male were significantly different before and after mating,among which Msep PR1,Msep PR3 and Msep PR5 genes were significantly higher before mating than after mating,while Msep PR2 and Msep PR4 genes were significantly higher after mating than before mating.The reason for the significantly increased Msep PRs genes in male after mating is unclear.A total of 21 OBP genes?including 2 common odor binding proteins,3 sex pheromone binding proteins,4 plus-c OBP genes,1 minus C OBP genes and 11 typical odor binding proteins?and 19 CSP genes were identified from the antennae transcriptome of male and female adults of M.separata.Quantitative analysis showed that 13 OBPs?Msep GOBP1,Msep GOBP2,Msep PBP1,Msep PBP2,Msep OBP1,Msep OBP3,Msep OBP6,Msep OBP6,Msep OBP7,Msep OBP8,Msep OBP10,Msep OBP11,Msep OBP14 and Msep OBP15?genes were specifically expressed in male and female antennae,among which 8 OBPs genes?Msep GOBP1,Msep PBP2,Msep OBP1,Msep OBP3,Msep OBP10,Msep OBP11,Msep OBP14 and Msep OBP15?were higher expressed in the antennae of female than that of male.Another four OBPs genes?Msep OBP2,Msep OBP4,Msep OBP9 and Msep OBP13?were highly expressed in testis,and the expression level was equal to or higher than male and female antennae.Msep OBP12 had a wide expression in different tissues.Msep OBP17 was highest expression in abdomen of female than the other tissues.We cloned an Msep OR13 gene of M.separata through transcriptome results,and further research were studied on this gene.The results showed that Msep OR13 was dominantly expressed in antennae of both sexs.Msep OR13 responded to 32 of 67 compounds.In addition,Msep OR13 also responded to eugenol at a low concentration of 10-9 M with an EC50 of 3.91×10-6 M.This high sensitivity of Msep OR13 suggests that Msep OR13 plays an important role in the insect's olfactory system.In this study,a large number of genes related to the olfactory sense of M.separata were obtained by sequencing the antennae of M.separata,which laid a foundation for the study of the mechanism of insect chemical communication system,and further analyzed its expression,providing a basis for future research on gene function. |
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