| Background:Aedes albopictus is widely distributed in different temperature zones in China,and is the main transmission medium of dengue virus(DENV),which poses a huge threat to human health worldwide.Understand the population genetic structure of Aedes albopictus under different temperature zones of our country,observe the growth and development indicators of the post-hybrid Aedes albopictus populations with large genetic differentiation among different temperature zones,and the differences in the susceptibility of hybrid offspring to DENV at different temperatures,explore the effect of temperature on the infection of dengue virus by different temperature zones and hybrid strains of Aedes albopictus.Analysis the changes in protein levels of Aedes albopictus ’ midgut after infection with DENV-2,and search for proteins related to DENV replication in the midgut,to provide an effective basis for the prevention and control of mosquito-borne diseases in our country.Methods:The genetic structure of nine different populations of Aedes albopictus in China was analyzed using COI molecular markers,and two populations with large differences in genetic differentiation were selected for hybridization,observe the The growth and development indexes of each group and dissected the midgut tissue.RT-qPCR was used to measure the dengue virus load in the midgut tissue of different groups at three different rearing temperatures of 23℃,28℃ and 33℃,and SPSS software was used to analyze the effect of time and temperature on the dengue virus load in the midgut.Proteomic analysis of the midgut of Aedes albopictus was performed with iTRAQ technology to find out the differentially expressed proteins between the infected group and the mock group.Among the differentially expressed proteins,cuticle protein and ubiquitin carboxy-terminal hydrolase proteins were verified by western blot,immunofluorescence,co-immunoprecipitation,etc.Results:1.The population genetic structure of nine different geographic strains of Aedes albopictus in different temperature zones was analyzed by COI gene,and it was found that the Aedes albopictus in Sanya showed greater genetic differentiation compared with other geographic strains.2.The groups of Beijing and Guangzhou were higher than the the hybrid groups in terms of hatching rate,pupation rate and fledging rate,while the hybrid groups were influenced by both sires and dams from the parents in terms of egg production rate,body length and resistance level.3.On the 3 and 7 days after infection with dengue virus,the virus copies in the temperature of 23℃ was significantly lower than that of the other two temperatures,but this difference narrowed after the virus established stable proliferation in the midgut at 14th days.The viral copies of Aedes albopictus in Guangzhou on the third day after infection with dengue virus was lower than that of the other groups at 23℃.4.A total of 3419 proteins were detected by iTRQA analysis,including 162 differential proteins,which were mainly concentrated in the metabolism of cytochrome P450,the constitution of cuticle and modified proteins.Among them,the cuticle protein can inhibit the replication of dengue virus and interact with DENV-E protein.And dengue virus infection can reduce the expression level of UCH L5-like.Conclusion:After hybridization of different Aedes albopictus geographical strains,the reproductiveability,developmental indicators,drug resistance levels and susceptibility to dengue virusare affected by both the male and female parents.The innate immune response of Aedes albopictus plays an important role in the mosquito antiviral process,in which cuticle protein and deubiquitination-related protein in the midgut tissue can interact with the virus and exerting the natural antiviral mechanism of mosquito midgut tissue. |
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