| This study used high-throughput sequencing,gene screening,quantitative reverse transcription(qRT-PCR)to analysis Harmonia axyridis ovary of the 2 day,3 days,5 days,7 days after emergence respectively,and make a further comment of the gene function prediction.The main conclusion of this paper is that:1.After Illumina Solexa deep sequencing,21.5 Gb of clean data were obtained from the transcriptome sequencing of the H.axyridis adult ovary.To obtain an overview of the ovary expression profile at the different developmental stages in H.axyridis adults,four expressed gene libraries(S1:22,364,281;S2:18,460,991;S 3:20,636,344;and S4:24,531,820)containing clean reads were obtained for analysis.12,985 unigenes were annotated by the GO classification system.The pathway involvement of 8144 unigenes identified in this study was predicted based on a comparison with the KEGG database.A total of 17,235,16,822,16,8635 and 28,186 unigenes were analysed with the KOG,Pfam,Swiss-Prot,and nr databases,respectively.Overall,28,558 unigenes were annotated.2.Comparing S2 with S1,962 genes were differentially expressed,including 134 up-regulated genes and 828 down-regulated genes.In S3 compared with S1,the expression of 524 genes was up-regulated and the expression of 810 genes was down-regulated.When the gene expression levels were compared between S1 and S4,4994 genes were up-regulated and 4646 genes were down-regulated,for a total number of 9640 genes3.26 genes related to ovarian development were screened out,among which the number of genes involved in the formation of polar granules was 1,belonging to 22 of the signal transduction mechanisms,with 3 genes related to the formation of vitellogenin.4.In order to verify the result of sequencing,20 differentially expressed genes were randomly selected for qRT-PCR.The results showed that the sequencing results were consistent with the results of fluorescence quantitative analysis.5.From the 26 genes of ovary development,we selected more important yolk protein genes to analyze the structure,and compared with other insects to analyze the status of the gene in the biological evolution. |
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