Coloring Mechanism Research Of Blue Flowers In Gentiana Sino-ornata Based On Transcriptome And Proteome

The flower of Gentiana sino-ornata is gorgeous blue,which is a good material for mining blue flower genes.This thesis found the important genes of Gentiana sino-ornata in anthocyanin synthesis pathway through combined analysis of the transcriptomics and proteomics,to explore the mechanism of blue floweration.And research on the key genes through molecular cloning and bioinformatics.The main results are as follows:1.Transcriptome identified a total of 108912 genes,with 105302 genes get function annotations.16846 up-regulated genes and 20603 down-regulated genes are identified,which are mainly involved in cell physiology and secondary metabolism biological processes,and some genes are unknown.The number of DEGs annotated to flavonoid biosynthesis and anthocyanin synthesis are 72 and 13 in H1-VS-H2,and 54 and 17 in H1-VS-H5,respectively.q PCR verified that the transcriptome data was reliable.2.The proteome identified a total of 2939 proteins,and they are annotated into 43 GO terms,124 pathways,and 24 KOG clusters,some proteins are unknown.912 up-regulated and 1061down-regulated proteins were identified,which were mainly involved in the secondary metabolite biosynthesis and flavonoid metabolism pathways.10 and 14 DEPs were annotated to anthocyanin synthesis in H1-VS-H2 and H1-VS-H5,respectively.3.There are 825 and 1247 DEPs matched with DEGs in H1-VS-H2 and H1-VS-H5,respectively,through the combined analysis of transcriptomics and proteomics.The key genes F3’5’H,5GT,5AT and cluster gene groups(including 3GT,UF3 GT,DFR,ANS,b HLH,GST)are identified,and we considered that they are the key gene group involved in the anthocyanins synthesis,regulation,and transport of the 5-acylated delphinidin in Gentiana sino-ornata flowers.We inferred the synthetic pathway and modification process of delphinidin,and analyzed the coloration mechanism of blue flowers based on these data.4.Obtained full-length c DNAs of Gs5 GT and Gs5 AT with complete open reading frames(ORF).Gs5 GT has a total length of 1465 bp and an ORF of 1365 bp and encodes 455 amino acids.Gs5 AT has a total length of 1594 bp and an ORF of 1422 bp and encodes 474 amino acids.According to bioinformatics analysis,Gs5 GT belongs to UDP-glucosyltransferase;Gs5AT belongs to anthocyanin glycosyltransferase.Gs5 GT and Gs5 AT are highly expressed in the flower and are highest expressed in the fourth stage,followed by leaves and rarely expressed in root and stem.5.Obtained a 1756 bp Gs5 AT promoter sequence,including TATA-box,CAAT-box,and other cis-acting element,lke light regulatory elements,plant hormone response elements,stress response elements,and tissue-specific expression regulatory elements,and it contains MYB,MYC(b HLH)transcription factor binding sites.