基于高通量测序454 GS FLX的丹参转录组学研究

Transcriptome characterization for Salvia miltiorrhiza using 454 GS FLX

本研究应用新一代高通量测序技术454GS FLX Titanium对2年生丹参根的转录组进行测序,研究其基因表达谱,挖掘其功能基因。获得46722表达序列标签(express sequence tags,EST),序列平均长度414bp,与Sanger测序的长度相当。所得序列与GenBank丹参EST合并拼接,获得18235条unigene,其中,454高通量测序发现了13980条新的unigene。数据库中的序列同源性比较表明,其中73.0%(13308条)与其他生物的已知基因具有不同程度的同源性。通过BLAST与Gene Ontology分析获得了可能参与丹参酮合成的序列27条(编码15个关键酶),参与丹酚酸合成的序列29条(编码11个关键酶),细胞色素P450序列70条,转录因子序列577条。454高通量测序技术作为药用植物功能基因组研究的重要手段可在丹参功能基因的发现中发挥重要作用,这些基因的发现为丹参酮和丹酚酸类化合物生物合成研究奠定了基础,同时也为丹参的转录组研究提供了基础数据。

To investigate the profile of gene expression in Salvia miltiorrhiza and elucidate its functional gene,454 GS FLX platform and Titanium regent were used to produce a substantial expressed sequence tags （ESTs） dataset from the root of S. miltiorrhiza. A total of 46 722 ESTs with an average read length of 414 bp were generated. 454 ESTs were combined with the S. miltiorrhiza ESTs from GenBank. These ESTs were assembled into 18 235 unigenes. Of these unigenes,454 sequencing identified 13 980 novel unigenes. 73% of these unigenes （13 308） were annotated using BLAST searches （E-value ≤ 1e-5） against the SwissProt,KEGG,TAIR,Nr and Nt databases. Twenty-seven unigenes （encoding 15 enzymes） were found to be involved in tanshinones biosynthesis,and 29 unigenes （encoding 11 enzymes） involved in phenolic acids biosynthesis. Seventy putative genes were found to encode cytochromes P450 and 577 putative transcription factor genes. Data presented in this study will constitute an important resource for the scientific community that is interested in the molecular genetics and functional genomics of S. miltiorrhiza.