Bioactive Substance Accumulation And Transcriptome Analysis Of Gardenia Jasminoides Fruit

Gardenia jasminoides,perennial evergreen shrub of genus Gardenia of Rubiaceae family,Its'fruit is multifunctional as traditional Chinese medicine and widely used in the pharmaceutical,food,cosmetic and dye.In this study,we observed the changes of chlorogenic,geniposide and crocin-1 contents in G.jasminoides(Linhai No.1)at different development stages.To figure out the accumulate regulation of the three bioactive substance.In addition,By transcriptome sequencing the fresh fruits at three vital development stages of G.jasminoides(60,120 and 180 DAF),Comprehensively analysed expression patterns key genes of bioactive substance synthesis and their pathway,We have find out the molecular mechanism of three bioactive substance' synthesis,chlorogenic acid,geniposide and crocin-1 synthesis-relate genes are confirmed meanwhile.This study enriches the transcriptome database of G.jasminoides,and provides important theoretical basis for the development and utilization of chlorogenic acid,geniposide and crocin-1 in G.jasminoides',as well as further research on high quality fruit of G.jasminoides resources with molecular markers and genetic breeding.The major conclusions are as follows:(1)G.jasminoides development characteristics.The growth trends of the G.jasminoides's horizontal diameter,vertical diameter,dry weight and fresh weight were almost the same,and they all showed a change from fast to slow.The color of the G.jasminoides gradually changes from blue-green to orange-red.With the expansion and maturity of G.jasminoides,there are different dynamic rules for the accumulation of three biologically active substances in the fruit:chlorogenic acid,geniposide and crocin-1:chlorogenic acid Accumulation showed a "first rise and then fall" pattern,the content reached a maximum of 0.25%at 120 DAF,and decreased to 0.17%at 180 DAF;crocin-1 showed a continuous growth accumulation pattern of"fast first and then slow",that is,from flowering Afterwards,the highest content reached 1.11%when it continued to accumulate to 180 DAF;the accumulation of geniposide showed a "first fall and then rising" pattern,with the highest content at 60DAF heing 2.04%,and at 120 DAF it fell to a minimum of 1.85%.and then slowly increased To 1.87%.(2)The correlation between the time and the main bioactive substances of the G.jasminoides found that 120 days after flowering(approximately mid-September),the content of chlorogenic acid was highest in G.jasminoides;As the fruit matures,,the content of crocin-1 in G.jasminoides is the highest in 180 days after flowering(approximately mid-November);the content of geniposide is highest in G.jasminoides in 60 days after flowering approximatelyt mid-July).Based on the appearance,shape and intrinsic quality of the fruit,the best harvesting time of chlorogenic acid,crocin-1 and geniposide in G.jasminoides were determined as mid-September,mid-November and mid-July,respectively.(3)Transcriptome sequencing of G.jasminoides at three different developmental stages,and bioinformatics methods such as sequencing evaluation and gene annotation were used to analyze the biosynthesis of chlorogenic acid,crocin,gardenia in G.jasminoides.After assembly,a total of 384421 Transcript and 298869 Unigene were obtained,with an average length of 836 bp and 510 bp,respectively.The functional comparison of Unigene in the Nr protein database,Swiss prot protein database,Pfam database,GO protein database and KEGG database were 152647(51%),92529(31%),91287(31%),23358(8%)and 46940(16%)Unigene sequences were successfully annotated.Based on the BLAST alignment of the public protein database and the functional group annotation of KEGG Pathway,the Unigene sequence and annotation information of key enzymes in metabolic pathways such as chlorogenic acid,crocin,geniposide,etc.Based on the valuable information of these key enzymes,schematic diagrams of the pathways related to effective components accumulation were established,and the effective components accumulation and metabolism mechanism of G.jasminoides was revealed.The biosynthesis process of gardenia fruit chlorogenic acid,crocin,and geniposide was systematically described.(4)The DE-Seq technology was used to analyze the differential genes between different developmental stages of the Unigene produced by the transcriptome analysis of Gardenia fruit.661 and 229 were found between 60 and 120 DAF,60 and 180 DAF and 120 and 180 DAF,respectively.Up-regulated with 55 differential genes,down-regulated with 1548,446 and 19 differential genes,and annotated through the KEGG metabolic pathway to determine the dynamic regulation pattern of related enzymes in the biosynthetic pathway of chlorogenic acid,crocin,geniposide.Combined with the comprehensive analysis of the dynamic changes of physiological metabolism of key enzymes in G.jasminoides,the improvation of chlorogenic acid,crocetin and geniposide genes was determinated by HCT,LCYB,and GPPS,respectively.