Expressional And Functional Characteristics Of Chemosensory Proteins AipsCSP8,AipsCSP9 And AipsCSP10 In Agrotis Ipsilon(Lepidoptera:Noctuidae)

The black cutworm moth Agrotis ipsilon(Hufnagel)(Lepidoptera:Noctuidae)is a worldwide agricultural pest.For a long time,the prevention and control of Agrotis ipsilon has been mainly based on chemical control.However,due to its strong migration ability and rapid development of drug resistance,the control effect of traditional pesticides has been greatly reduced,and the degree of harm has become increasingly serious.And chemosensory proteins play a vital role in the olfactory mechanism of insects.They can combine chemical information substances or sex pheromones and transport them.They can participate in the process of insects looking for hosts and feeding.Therefore,the study of chemosensory protein function is greatly significant for the research and development of new efficient,environmentally friendly and green prevention methods and technologies.In order to clarify the functions of AipsCSP8,AipsCSP9 and AipsCSP10 and their new functional characteristics that may evolve in the process of long-term control of Agrotis ipsilon,the research mainly used a series of molecular biology technologies and methods to systematically express expression and functional characteristics of 3 chemosensory proteins in Agrotis ipsilon at different levels respectively.It is hoped that these will provide environmental new target approaches for effective prevention and control of Agrotis ipsilon in the future.The main results of this study are as follows:1.Tissue expression profiles of AipsCSP8,AipsCSP9 and AipsCSP10 genes in male andfemale adultsUsing qRT-PCR experiments to determine the tissue expression profiles of AipsCSP8,AipsCSP9 and AipsCSP10 genes in Agrotis ipsilon,the results show the expression level of AipsCSP8 was the highest in the leg and labial palp of male adults,followed by in the accessory gland and the antenna of male adults.While in female adults,the expression level of AipsCSP8 was the highest in the antenna,followed by in the leg,pheromone gland and labial palp.AipsCSP9 and AipsCSP10 genes showed specific expression in male and female adults,and the expression levels of those genes in antennae were significantly higher than in other tissues.2.Spatio-temporal expression profiles of AipsCSP8,AipsCSP9 and AipsCSP10 genes in male and female adultsUsing qRT-PCR experiments to determine the spatio-temporal expression profiles of AipsCSP8,AipsCSP9 and AipsCSP10 genes in male and female Agrotis ipsilon antennae,the expression levels of AipsCSP8 gene in the antennae of males and females reached the peak on the 1st day before emergence and on the day of emergence,respectively.The expression levels of AipsCSP9 and AipsCSP10 genes gradually increased from before emergence,and gradually reached the peak after emergence.The expression levels of AipsCSP9 gene from 1 day after emergence to 6 days after emergence were no significant difference.The expression levels of AipsCSP10 gene from 0 day after emergence to 6 days after emergence were no significant difference.3.The expression patterns of AipsCSP9 and AipsCSP10 in different antennal sensillaIn situ hybridization experiments and immunohistochemical experiments showed that AipsCSP9 and AipsCSP10 were expressed in sensilla trichodea of both female and male antennae,and no co-expression of the two CSPs was found.4.Binding characteristics of AipsCSP8、AipsCSP9 and AipsCSP10The results of fluorescence competitive binding assay showed that AipsCSP8 had strong binding capabilities with dibutyl phthalate,phenylacetaldehyde and avermectin,AipsCSP9 had strong binding capabilities with dibutyl phthalate,avermectin,tebufenozide,acetamiprid,thiamethoxam,AipsCSP10 had strong binding capabilities with(Z)-5-Decen-1-ol,(E)-11-Tetradecen-1-ol-acetate,tebufenozide,acetamiprid.The results indicate that AipsCSPs not only have functions in identification of sex pheromones and host plant volatiles,but with the use of chemical pesticides,they may also play important roles in the identification of chemical pesticides.To explore the expression and functional characteristics of 3 chemosensory proteins AipsCSP8,AipsCSP9 and AipsCSP10 in Agrotis ipsilon,the study determined their tissue expression profiles and spatio-temporal expression profiles,and clarified AipsCSP9,AipsCSP10 expression pattern in antennae of male and female adults.At the same time,the experiment was performed,by adding chemical pesticides as test ligands on the basis of using sex pheromone and plant volatiles as test ligands,to find the possible functions of AipsCSP8,AipsCSP9 and AipsCSP10 in the recognition of external chemical stimuli.The study suggests AipsCSP8,AipsCSP9 and AipsCSP10 may be involved in the olfactory behavior of Agrotis ipsilon and play roles in the perception of chemical pesticides.And among them,AipsCSP8 may also participate in the taste recognition process of Agrotis ipsilon.