Study On The Selection Of Blue Flower Germplasm And Their Flavonoid Related Compounds In Petals

Blue flowers give people a cool and comfortable feeling,because some of the valuable flowers on the market lack of blue flowers,the blue flower is the hot spot and difficulty in research.In this study,the author combines the phytochemistry and molecular biology to analyze the flavonoids content of blue flowers.And then screen a blue plant(Gentiana sino-ornata),sequence the transcriptome,the quality and the results of the transcriptome with the Illumina HiSeqTM 2000 platform.Cording to the functional notes,explore color formation of genes,differentially expressed genes,and the expression of a part of the genes to verify.1.Make a phenotypic determination of three blue flowers plants use the colorimeter.Measure the anthocyanin content and intermediate metabolites with HPLC-PAD;Measure the composition of anthocyanins with HPLC-ESI-MS.The results are as follows:(1)The linum pernne b*value is-23.9,The anthocyanin component delphinidin,cyanidin,malvidin.The three anthocyanins are all acylated.The delphinidin content accounted for 60%of total anthocyanin;(2)The G.veitchiorum Hemsl.b*value is-22.1,The anthocyanin component Pelargonidin,cyanidin and delphinidin.Only the delphinidin was acylated;(3)The Gentiana sino-ornata b*value is-36.82,The anthocyanin component delphinidin and pelargonidin.The two anthocyanins are all acylated,The delphinidin content accounted for 95%of total anthocyanin.Comprehensive analysis of three plants,the Gentiana sino-ornata contains 8 kinds of anthocyanin,and all acylated..So we selet it to further study.2.Take the three flowering stages of gorgeous gentian:Taking the first stage(H1-R)of petals without tinting,the second stage(H2-R)of petals with half tinting and the fully open stage(H5-R)of the fifth stage as plant materials.The total RNA was extracted using the base of the Illumina HiSeqTM2000 platform for transcriptome sequencing.Assemble the results of the transcriptome sequencing,gene function annotation,and so on,then construct the transcriptome database.After sequencing,the Unigene was 72770,the average length was 6 bP,and the length of Unigene sequence was within the range of 617.20 bp.Among them,39330 genes were annotated,accounting for 54%of the total.There are 39169 comments in the Nr database,in the SwissProt database to get 23418 notes Unigene and in the KEGG database to get 23418 notes Unigene,in the COG/KOG database to get 10649 notes Unigene.Through the functional annotation of Unigene,there were 127 genes involved in the anthocyanin synthesis,of which 6 were up-regulated and the other 13 were significantly down regulated.3.The study of Gentiana sino-ornata three flowering stage of gene expression was verified on 9 Unigene(Unigene0037993,Unigene0023341,Unigene0014078,Unigene0055940,Unigene0059995,Unigene0053146,Unigene0054587,Unigene0051274,Unigene0012123)were validated by quantitative PCR,the results found that changes in the rate of the same amount of fluorescence quantitative PCR results and transcriptome sequencing expression,transcriptome sequencing results proved reliable.4.The expression patterns of 9 genes were analyzed,The expression patterns of these 6 genes Unigene0037993,Unigene0014078,Unigene0055940,Unigene0054587,Unigene0051274 and Unigene0012123 increased firstly and then decreased,the expression patterns of these 3 genes Unigene0023341,Unigene0059995 and Unigene0053146 showed decreased and then increased.Among them,the annotation of FLS and DFR expression of the two genes in the opposite trend,sharing the same substrate,showing a competitive relationship.