| The study is the research contents of National Forestry Administration for ’948’project and the Central University Innovation Project.In this experiment, Iris sanguorsinea and Iris sanguinea f. albiflora are the research materials.The study is based on the transcriptomics date analyzing of two flower design and flowering period plant material, as well as the controller and analyzing existed in physio-logical indicators different growing period, aims to study two color Iris sanguinea differential expression of anthocyanin synthesis and color variation related enzymes associated structural genes, the results is that:(1)The results of Total Clean Reads of Iris sanguinea and Iris sanguinea f. albiflora was 53,661,436,55,020,892, number of assembled Unigene is 69,690 and 71,943, and the average length is 496 nt and 521nt. After the combination, there are totally 67,399 Unigene and every length is more than 200nt. Therefore, transcriptome sequencing and assembly quality is good.(2) There are a total of 44,423 Unigene compared to public database, where Nr (99.34%), Nt (73.03%), GO (68.78%), KEGG (58.58%) and COG (37.11%). Then the highest rate of model species is grapes (21.1%), followed by rice (9.2%).(3) The significant genes expression in Iris sanguinea and Iris sanguinea f. albiflora are totally 10,493, of which the Iris sanguinea f. albiflora has 6749 up-regulated expression, 3744 down-regulated expression genes.(4) These categories have the most significant different gene expression Unigene in the GO database:cell, cell part, organell, cellular process and metabolic process; These categories have the most significant different gene expression Unigene in the COG database:general function prediction only, transcription; there are significant different genes contrasted to 121 pathways KEGG, in which the most deferentially expressed five paths genes are metabolic pathways, Biosynthesis of secondary metabolites, Endocytosis, Glycerophospholipid metabolismpathway and Ether lipid metabolism.(5) Through the classic screening of KEGG pathway, Anthocyanin biosynthesis (pathway ID for ko00942) has found, and 13 Unigene where in the NCBI accession of 11. Significantly different genes in both color Iris sanguinea anthocyanin biosynthetic pathway is a commen t to the number of genes for five. Among those, ID close related to Iris sanguinea and Iris sanguinea f. albiflora are 5GT and 3GT, compared with Iris hollandica, there are down-regulated expression genes in Iris sanguinea f. albiflora.(6) Based on the anthocyanin biosynthesis pathway, transcription differences between the two sets of data are used to suit sanguinea, and 17 anthocyanin biosynthesis gene structure are filtered, compared with Iris are CHS, ANS, F3H,5GT and 3GT. compared with Iris germanica is CHS, compared with Iris hollandica are ANS, F3H,5GT,3GT.(7) There are 7 gene of anthocyanin biosynthesis with the highest rate of gene transcription in the screening group selected, take XM 003602497.1 as reference gene for real-time pcr. The results of seven selected genes:check CHS, F3H, ANS, F3’H, F3’5’H, DFR and Real-time quantitative expression of the gene results transcriptome sequencing.6 genes are consistent with a phase change. Among which genes Iris sanguinea f. albiflora unregulated compared with sanguinea is:F3H, F3’H, F3’5’H, DFR, reduced expression is:ANS, CHS; quantitative results of CHI Real-time and transcriptome sequencing are opposite.(8) The results related to physiological indicators of color are:content of Iris sariguinea f. albiflora is close to zero, anthocyanin content Iris sanguinea changes with the developmental stages showing a downward trend after the first rise in the early opening of the highest value; two kinds of Iris sanguinea color change in the activity of the four developmental stages of PAL enzyme varies developmental stages and weakened; Iris sanguinea f. albiflora CHI activity increases gradually reduce developmental stages, and the CHI activity of various developmental periods are significantly higher in Iris sanguinea, Iris sanguinea CHI activity changes at first and then increase decreasing; two color Iris sanguinea soluble sugar content in four developmental stages show a downward trend, and the trend is opposite to soluble protein content; PAL activity, CHI activity, soluble sugar content and anthocyanin synthesis are of positive correlation in a certain degree, soluble protein content and anthocyanin content are negatively correlated.(9) Iris sanguinea f. albiflora in CHS gene showed significantly reduced expression,5GT, 3GT, ANS F3H gene expression are significantly up-regulated genes which can be used as the main structural variation in the kinds of colors Iris sanguinea a research. |
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