Transicriptome Analysis Of Eleutherococcus Senticosus And Cloning And Functional Resaerch Of FPS Promoter

In order to obtain the transcriptome data of Eleutherococcus senticosus,its differentially expressed genes of triterpenoid saponin in high-content group(B)and the low-content group(M),and farnesyl pyrophosphate synthase gene(FPS)full-length.The second-generation of Illumina Hwaseq 4000 sequencing platform,bioinformatics methods,PCR and thermal asymmetric interlaced PCR(TAIL-PCR)were used to conduct researches.The main results were achieved: 1)The transcriptome data of E.senticosus was 8.34 Gb and included 77087 Unigenes;the total length and average length of transcriptome were 52940804 nt and 687 nt respectively;and the length of N50 was 1207 nt.Compared with 7 gene databases,29348 Unigenes obtained annotation information,and the transcriptome data of E.senticosus was the highest similarity with grapes,its transcriptome data could be classified into 55 GO terms and 24 COG terms,as well as involved 116 KEGG pathways.Meanwhile,it was predicted that there were 59098 CDSs,5028 SSRs,and 162831 SNPs.2)As a result,530 differentially expressed genes were selected,including 223 up-regulated genes and 307 down-regulated genes.Compared with GO,COG and KEGG,a total of 434 genes were annotated.3)The total length of FPS was 7952 bp and the A/T accounted for 64.29%.The transcription initiation site of FPS from E.senticosus was G,and it was located at 117 bp upstream of the initiation codon ATG.There were 12 exons and 11 introns in FPS DNA sequence and 120 cwas-acting elements in its promoter,25 types,core functional elements of 62 TATA-box and 24 CAAT-box,and 28 other important functional elements.After ligating the p CAMBIA1301 vector with FPS promotor and transfecting E.senticosus,it proved that the promoter had promoter activity.Conclusion: 1)For the first time,the transcriptome database of E.senticosus was established,and a large number of transcripts were obtained,which provided valuable genomic resources for the study of the molecular biology of E.senticosus.2)The differentially expressed genes between the high and low content of triterpenoid saponins were screened to provide reference for further study of triterpenoid saponin biosynthesis and accumulation.3)For the first time,the full length of FPS from E.senticosus was cloned,which laid the foundation for further study on the regulation of FPS expression in E.senticosus.