Key Genes Cloning And Expression Analysis In Pathway Of Terpenoids Metabolism Of Blumea Balsamifera L.DC

Blumea balsamifera L.DC belongs to compositae and it is a kind of perennial herbs.Blumea balsamifera L.DC was registered first in A.D.741(Tang dynasty).Blumea balsamifera L.DC is a very important Chinese traditional medicine,its roots,leaves were regarded as drug parts.Blumea balsamifera L.DC has lots of medical effects,such as invigorating the circulation of blood,curing arthritis,curing dysmenorrhea ect.And the Blumea balsamifera L.DC mainly distributes in Hai Nan,Gui Zhou,Guang Xi and so on.Recently,many studies of Blumea balsamifera L.DC were started or finished,but no any research refer to inner substantial mechanism of terpenoids synthesis and accumulation,the existing researching results cannot illuminate many key points,for example how to regulate the metabolism of terpenoids,how to evaluate gene expression level.In order in resolve those problem,our study group has sequenced transcriptome and clone key genes in terpenoids metabolic pathway,Evaluating gene expression level by q-PCR technology.Main researching respects are:1.Select the most excellent exogenous inducers by thinking about concentration of Left-handed borneol,enzymatic activity,and appearance change of leaves.Result demonstrated that 1mmol/L Methyl jasmonic acid(MeJA)was treated as optimum inducer.2.Transcriptome sequencing.Removing insignificant reads,983 415 36 reads of Blumea balsamifera L.DC were obtained.All of the 983 415 36 reads contained 847 738 540 1 nucleotides.After de novo splicing,100 341 Unigenes were obtained.The Unigenes were aligned in COG data base,searching result demonstrated that Unigenes were devided into 25 classes according to function.The Unigene GO functions can be broadly divided into biological processes,cellular components and molecular function categories of 43 branches.In KEGG database,the data in the transcriptome can be divided into 111 classes according to the metabolic pathway which included the biochemical pathway in plants—Pathogens interaction,terpenoid and steroid compounds synthesis,litlid metabolism,RNA degradation and so on.3.Clone the key genes about terpenoids synthesis and analyze the genes` information via bioinformatics way.4.Design special primers to study GPPS gene and TPS gene expression level in q-PCR technology.The consequence showed that GPPS and TPS increased their expression level after 24 h,GPPS and TPS decreased their expression level after 48 h.