Gene Expression Analysis And Biotechnology Application Of Key Enzymes Of Jasmine

Jasmine tea is praised by the masses of consumers in the mar ket,and the quality standard of jasmine tea mainly depends on the fragrance of jasmine.Jasmine is the typical nocturnal flower,with the characteristics of opening at night and releasing fragrance.The main aroma compounds of Jasmine are linalool,benzyl acetate and Alpha farnesene.Firstly,with the different proportions of the three essential oils in this research,it is clear that the proportion of ingr edients,from the sensory point of view,close to jasmine and its importance,and then,the further study on the aroma metabolism pathway of the main aroma components of jasmine was carried out,It was found that only when the three components are present and the mass ratio is about 12:5:8 is the closest to the jasmine flower,the order of importance:preliminary judgment for benzyl acetate,alpha farnesene,linalool.Benzyl acetate is an ester compound produced by benzene metabolism pathway,Alpha farnesene is a sesquite rpene compounds produced by terpenoid metabolic pathway,Linaloo 1 is alcohol compounds produced by terpenoid metabolic pathway,In this study,the key enzymes,as AACT,TPS,DXR,GGDS,FDS in the terpenoid metabolic pathway,and EPSPS,PAL,CNL,CHD,K AT1,BALDH,BEAT,ODO1,EOBII in the benzene ring metabolic pathway,were selected.Blast analysis of the genes homologous sequence which was related to the known function enzymes that contai n TPS,AACT DXR,GGDS,FDS,EPSPS,PAL,CNL,CHD,KAT1,BALDH,BEAT,ODO1,EOBII was used to Jasmine Transcriptome in our laboratory.Jasmine cDNA sequence was acquired that can be used to screen and analysis through Bioinformatics.The expression of target gene RT-PCR was showed that the high expression of genes at night related to key enzymes,TPS,DXR,GGDS,EPSPS,FDS,in the terpene metabolic pathway and key enzymes,EPSPS,CHD,BALDH,CNL,JsODO1,in the benzene metabolic pathway,during the day to low expression.These genes are conformed to biological learning i accordance with jasmine’s flower opening and releasing aroma atght.Among these key enzymes,especially the key enzymes,TPS,L R,GGDS,EPSPS,FDS,in the terpene metabolic pathway of coding genes have higher expression at night and n o expression during the day.It is indicated that these coding genes of key enzymes in the terpene metabolic pathway have a large effect on releasing arom from Jasmine.In this study,the coding genes of TPS,AACT,(?)S and FPS,which are the key enzymes of terpenoid pathway,w(?)cloned,The results were compared with those obtained by agar(?)gel electrophoresis,and the results were compared with those of the transcriptome data.Further study was done by the virus induce(?)gene silencing(VIGS)technique,Firstly,pTRV2e was used for s(?)le enzyme digestion,then the coding gene was connected with the pTRV2e virus vector by LIC reaction,currently connected successfully with pTRV2e-c58633 and pTRV2e-c3956,through the analysis of PCR,agarose gel and sequencing,it was proved that pTRV2e-c(?)633 and pTRV2e-c3956 were connected successfully,and no mutaion occurred,then pTRV2e-c58633 and pTRV2e-c3956 as well as(?)TRV2e enzyme empty vector and pTR V1 were transferred into Agn bacterium by freeze thawing method,testing the application of VIG(?)technology of jasmine aroma in the study.