Study On The Life History Of Common Sarcosaphagous Flies In Guiyang And Transcriptome In Pupal Stage

Objective: To observe the relation between temperature accumulated and life history of Hemipyrellia Ligurriens,Muscina Stabulans and Boettcherisca Formosensis under the indoor with natural conditions in summer in Guiyang,and to discuss the differential expression of Boettcherisca Peregrine pupa’s transcriptome in three different development times,so as to provide experimental data for further studies on the differential genes in the transcriptome of Boettcherisca Peregrine pupa in various stages.Methods: 1.Put the corpses of rabbit in Guizhou Medical University and Guizhou Police Officer Vocational College respectively to trap three common sarcosaphagous flies,namely Boettcherisca Formosensis,Muscina Stabulans and Hemipyrellia Ligurriens,feed them until they spawn,and then observe the durations of the three sarcosaphagous flies’ developmental stages under the natural conditions of summer in Guiyang while record the temperature and humidity.2.Put the corpses of rabbit for experimental use in the outdoor area of Guizhou Medical University,and gather the Boettcherisca Peregrine on the corpse for artificial feeding.After they gave birth to larva and pupated,collect the fly pupae every 24 hours and two grains of fly pupae for one time until they emerged to adults.Choose the fly pupae collected on the first,the fifth and the tenth day to extract the total RNA,and then carry out sequencing and library construction.Analyze the gene differential expression by using the software of edgeR,screen the differential gene for real-time fluorescence quantification.Apply Kyoto Encyclopedia of Genes and Genomes(KEGG)metabolic pathway analysis to the known genes(Unigene),and implement simple sequence repeat(SSR)detection.Results: 1.The developmental duration of Boettcherisca Formosensis in June,July and August is 17.00 d,15.19 d and 15.7d,respectively;the developmental duration of Muscina Stabulans in June and July is 16.13 d and 13.9d,severally;and the developmental duration of Hemipyrellia Ligurriens in July is 13.06 d.The developmental durations of Muscina Stabulans and Hemipyrellia Ligurriens in July are similar;while the developmental durations of Muscina Stabulans in June and July are somewhat different from those of Boettcherisca Formosensis in June,July and August.The essential data about the total developmental durations and accumulated temperature of the three kinds of sarcosaphagous flies are calculated.2.Acquire the expression level of sample genomes in different groups,and capture 38,727 Unigenes totally.The transcriptome results of fly pupae in different stages were used to construct database and compared in pairs.The total number of differential genes was 5,283.Construct visual scatter diagram and volcano plot for differential expressed genes,apply gene ontology(GO)classified statistics and enrichment analysis to differential genes and screen tyrosine hydroxylase(TH)gene and CBM-14 gene with distinctive differentiation for real-time fluorescence quantification,the results of which were in consistent with those of sequencing.In the transcriptome of boettcherisca peregrine,the total number of Unigenes and metabolic pathways that were annotated was 8,856 and 336,severally.Through SSR,it can be found that SSR loci among Unigene account for 22.23%.The repetition of mononucleotide occupied the highest proportion,followed by the repetition of trinucleotide.Among different repetitive primitive types in SSR,A/T appeared with the highest frequency,followed by AAC/GTT and AC/GT.Conclusion: 1.Information about the life history of three kinds of sarcosaphagous flies in Guiyang,including Hemipyrellia Ligurriens,Muscina Stabulans and Boettcherisca Formosensis is geted.2.The transcriptome database about the three different developmental stages of boettcherisca peregrine in Guiyang is constructed.3.The expressions of TH gene and CBM-14 gene are significantly differential in three developmental stages of boettcherisca peregrine.