Olfactory Gene Expression Of Culex Pipiens Pallens And Culex Pipiens Molestus Based On RNA-Seq

Culexpipienspallens and Culexpipiensmolestus are two subspecies of the same species distributed in the Culexpipiens complex.Culexpipienspallens is a non-self-fertilizing insect.The first reproductive nutrient cycle needs to suck the blood of the host animal before it can be spawning.Culexpipiensmolestus is a self-fertile insect.In the first reproductive nutrient cycle it can spawn to progeny without bloodsucking.Culexpipienspallens and Culexpipiensmolestus exhibit evolutionary differentiation.The olfactory system is an information transmission system in which insects perceive the surrounding environment,find spawning sites,and search for host animals.Therefore,the evolutionary differentiation of Culexpipienspallens and Culexpipiensmolestus also has evolutionary differentiation in the gene expression level of the olfactory system in theory.By studying the differential expression of transcriptome and olfactory genes,this study not only provides a basis for revealing the molecular biological mechanism of evolutionary differentiation,but also provides a reference for the development of new anti-mosquito agents with targeted olfactory blocking.In this study,before the blood-sucking of Culexpipienspallenscoquillett,after the blood-sucking of Culexpipienspallenscoquillett,before the blood-sucking of Culexpipiensmolestus,and after the blood-sucking of Culexpipiensmolestus,the RNA-Seq technique and bioinformatics method were used to study the antennal transcriptome and olfactory genes before and during blood-sucking.The difference in expression between Culexpipienspallenscoquillett and Culexpipiensmolestus,on the one hand,clarifies the evolutionary differentiation of the transcriptome and olfactory genes of Culexpipienspallenscoquillett and the olfactory organs of Culexpipiensmolestus.On the other hand,screening may be related to blood-sucking and searching for spawning water.Search for the olfactory genes associated with the host animal.Transcriptome analysis showed that after the blood-sucking of Culexpipienspallenscoquillett,there were 4,162 gene expression differences in the transcriptome,of which 2390 were significantly up-regulated,and 1772 were significantly down-regulated.After Culexpipiensmolestus,a total of 1755 genes were significantly different in the transcriptome,with 493 significantly up-regulated and 1262 significantly down-regulated.The difference in the number of genes differentially expressed between Culexpipienspallens and the transcriptome of Culexpipienssinensis was extremely significant.The results of GO enrichment showed that the genes differentially expressed by Culexpipienspallenscoquillett and Culexpipiensmolestus were only the same functional group of RNA binding,ncRNA and ion transmembrane transporter activity,and the other 54 functional groups were different.The results of KEGG enrichment showed that Culexpipienspallenscoquillett and Culexpipiensmolestus were not significantly enriched in the same pathway after blood-sucking.Before blood-sucking,the expression level in the antennae of Culexpipienspallenscoquillett was significantly higher than that of the strains of Culexpipiensmolestus.It was mainly related to enzyme activity,cell stress response,signal transduction and other related functions;It is related to functions related to NADH activity,peptide metabolism,and organic nitrogen synthesis.After blood-sucking,the expression level of Culexpipienspallenscoquillett was significantly higher than that of Culexpipiensmolestus.The genes were mainly enriched in ribosome structure,serine activity,peptide metabolism and other related functions;the gene significantly lower than that of Culexpipiensmolestus was only enriched in ion transport of related features.After the blood-sucking of Culexpipienspallenscoquillett,there were 12 significantly elevated olfactory genes OBP,and there was no significant down-regulation of OBP.After Culexpipiensmolestus,there were three significant up-regulations of OBP,of which OBP23 and OBP44 were the same as Culexpipienspallenscoquillett.five of the significantly down-regulated olfactory OBP.Before the blood-sucking,the OBP expression of Culexpipienspallenscoquillett was significantly higher than that of the Culexpipiensmolestus,which was significantly lower than that of the Culexpipiensmolestus.After blood-sucking,OBP of Culexpipienspallenscoquillett was significantly higher than 13 of Culexpipiensmolestus,and significantly less than 3 of Culexpipiensmolestus.There are 5 significant OR-related genes down-regulated by Culexpipienspallenscoquillett,the most notable being CPIJ013183.Only one of the significant upward adjustments is CPIJ003158.After murdering Culexpipiensmolestus,three genes were down-regulated,most notably CPIJ001069;there was no significant up-regulation of genes,of which CPIJ003158 was expressed in contrast to Culexpipienspallenscoquillett,and Culexpipiensmolestus was significantly down-regulated.Before blood-sucking,the expression of OR-related genes in Culexpipienspallenscoquillett was significantly higher than that of four Culexpipiensmolestus,among which CPIJ005662 was the highest;5 was significantly lower than that of Culexpipiensmolestus,and the lowest was CPIJ006217.After blood-sucking,Culexpipienspallenscoquillett was significantly lower than the three genes that Culexpipiensmolestus,the lowest being CPIJ006984.The expression level of Culexpipienspallenscoquillett CPIJ003158 before bloodsucking was significantly lower than that of Culexpipiensmolestus.Differential analysis of transcriptome and olfactory gene expression showed that the gene expression of Culexpipienspallenscoquillett and Culexpipiensmolestus showed significant evolutionary differentiation.OBP23 and OBP24 may be the same target genes for Culexpipienspallenscoquillett and Culexpipiensmolestus after blood-sucking to find water for spawning.CPIJ013183 may be an important olfactory gene for Culexpipienspallenscoquillett to find a host animal to suck blood.CPIJ001069 may be the main functional gene for Culexpipiensmolestus to search for host animals.CPIJ003158 may be an important gene for Culexpipiensmolestus to search for host animals.The above bioinformatics analysis prediction results need further verification.RT-qPCR verification showed that expression of OBP31、OBP44、OR65 and GPCR4 of Culexpipiensmolestus were significant up regulated post blood feeding,and significant higher than Culexpipienspallens.They maybe regulate Culexpipiensmolestusbehavior specialized.