| Wohlfahrtia magnifica are the main pathogen that invades the vagina of bactrian camels and causes vaginal myiasis,which has a great harm on the reproduction and health of camels.On the basis of previous studies,we used the ultra depth imager to identify the terminalia of male and female Wohlfahrtia magnifica,and then observe the morphology of male and female antennae by scanning electron microscope.We used the male and female antennae as samples for transcriptome analysis and quantitative real time PCR(q RT-PCR)analysis of olfactory related genes.The results showed that: 1.The structure of terminalia is difference between male and female,the female flies has serrated end edges in the caudal end of abdominal segment and its terminalia is concave and the anal papilla is grayish white.To the male flies,its end edges of the caudal end of abdominal segment is smooth and its terminalia is convex,the dark yellow saccular process is shaped like a spoon swollen.2.The antennae of Wohlfahrtia magnifica are typical awn-like tentacles,and the sensilla types of male and female adults are the same.The antennal sensilla can be divided into 4 categories.They are divided into: Sensilla trichomes,Sensilla basiconica,Sensilla chaetica,and Sensilla coeloconica.Different sensilla have different functions.3.The experiment performed transcriptome sequencing on the antennae of the Wohlfahrtia magnifica.From the sequencing results,it can be seen that the GC content is more than 29%,the Q20 content is more than 97%,and 50387 unigenes were assembled,and the assembly results were stored in 5 databases.For comparison,after the transcriptome assembly of Unigene sequence,21018 annotations in NCBI’s Nr database.After GO function annotation,it is found that 14935 cells account for 79.20% and cell part14934 accounts for 79.19%.The ingredients account for the largest proportion,which is compatible with the antenna function.KEGG analysis found that signal transduction accounted for a large proportion in genetic information processing,indicating that signal transduction and pathways play an important role in the antennae of Wohlfahrtia magnifica.According to the analysis of female and male differential gene expression,6008 unigenes were significantly different between the two groups,2211 unigenes were up-regulated and 3797 unigenes were down-regulated.Highly expressed olfactory-related genes Obp2,Obp71,Obp24,Or10 a,Or30a,SNMP1 were screened out for verification experiments.4.Fluorescence quantitative PCR results show that SNMP1 gene is expressed in various tissues,and the expression level in female and male antennae is significantly higher than that in other tissues,and there is no significant difference in the expression level in female and male antennae(P>0.05).The expression of Obp24 gene in the antennae,head and leg is significantly higher than that of the thorax and abdomen,and there are significant differences in female and male expression.The expression in female antennae is extremely significant than that in male(P<0.01),the expression of Obp71 gene in antennae and male fly head.It is extremely significantly higher than the head,thorax,abdomen and feet of female and male flies(P<0.01),the expression levels of Or10 a and Or30 a genes in male and female antennae are significantly different and significantly higher than other tissues(P<0.05),the difference in expression levels of Obp2 gene in male and female antennae is significant(P<0.05).In this study,we obtained the morphological structure of male and female antennae and sequenced the transcriptome of antennae,and identified the expression of olfactory related genes,which laid a molecular foundation for further study of gene function and olfactory sensing mechanism. |
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