Identification Of Sex Pheromone And Cloning And Tissue Expression Pattern Of Odorant Binding Protein Genes In Athetis Dissimilis Hampson

Athetis dissimilis Hampson belongs to lepidoptera noctuidae and becomes dominant in agricultural pests recently,endangers crops such as corn,wheat,cotton and so on.Insect sex pheromone has many advantagages,such as specificity,non-pollution and harmless of natural enemy and useful organisms,and thus has been applied in control of many insect pests.However,sex pheromone of Athetis dissimilis Hampson was barely studied.In the present study,a sex pheromone component of Z9-14:OH was firstly identified from the female pheromone gland of A.dissimilis,by GC-EAD and GC-MS.Then,the EAG response and dose effcet of male antenna to Z9-14:OH and a pheromone analogue Z9,E12-14:OH was tested.Thirdly,the baits were prepared based on the EAG results,and the field trapping experiment was conducted.Finally,to investigate olfaction mechanism,2 general odorant binding protein(GOBP)genes and 3 pheromone odorant binding protein(GOBP)genes were obtained by analyzing antenna transcription data and PCR varidation,and their tissue expression patterns were determined by RT-qPCR.The main results were as follows:1.Identification of sex pheromone in Athetis dissimilisGC-EAD results revealed that male antenna had a strong EAG response on the female pheromone gland extraction,and a sex pheromone component was identified as Z9-14:OH by GC-MS.Dose effect on EAG response of Z9-14:OH and the analogue Z9,E12-14:OH was tested.Results show that EAG responses increased with dose within the test range(0-1000 ng)for both compounds;EAG responses by Z9-14:OH at doses of 100 ng and above were significantly higher than that the solvent control,while the significant EAG response by Z9,E12-14:OH was only observed at the dose of 1000 ng.Field trapping experiments was conducted using rubber baits(total dose of sex pheromone components was 0.5 mg per bait)consisting of different ratios of Z9-14:OH and Z9,E12-14:OH(10:0,0:10,1:9,3:7,5:5,7:3,9:1).Results showed that the baits at ratios of 0:10 and 1:9 caught highest number of male moths.Considering the low A.dissimilis population in the fields,this result needs further verification.2.Identification and tissue expression pattern analysis of 2 GOBPs and 3 PBPs in A.dissimilisTwo GOBP genes and 3 PBP genes were identified by antennal transcriptomic anaylysis and RT-PCR,and the tissure expression profiles of these five OBP genes were determined by RT-qPCR.Results show that all 5 OBP genes expressed specifically in male and female antennae and did not express in other parts including head,chest,abdomen,feet and wings.In comparison between sexes,the expression levels of GOBP1,GOBP2 and PBP3 gene in the female antennae were 2.33,1.75 and 1.05 times higher than that of males,respectively;while PBP1 and PBP2 gene in male antennae were 1.84 and 1.14 times higher than that of females,respectively.These bases provide for further elucidation of the function of these genes and further the molecular mechanisms olfaction in A.dissimilis.