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目的:对天麻进行转录组学分析,从转录组水平阐释天麻次生代谢产物含量与相关酶活性的关系。方法:采用Illumina Hiseq 4000对新鲜天麻及采后敞放5 d天麻进行转录组测序分析,通过与基因数据库比对分析注释和发现差异表达基因。结果:新鲜天麻及采后敞放5 d天麻分别获得了18 772 600、19 353 329条Clean Reads,Trinity组装后得到63 455条Unigene,通过与NR、Swiss-Prot、GO、COG、KOG、KEGG数据库比对,最终获得18 913个有注释信息的Unigene。天麻采收放置5 d后有953个基因转录表达量上升,1 876个基因转录表达量下降,合计2 829个差异表达基因。634条差异表达基因被COG注释到25个功能类别中,612条差异性表达基因被KEGG注释归属于50条代谢通路。天麻转录组差异表达数据中共11个关键基因被KEGG数据库苯丙烷类合成途径注释,与新鲜天麻相比,放置5 d的天麻CM、PDT、PAL、4CL、CHS、F5H、F3’M基因表达量下降,C4H、CCR表达量上升,CAD表达量既有上升又有下降。结论:本研究获得了较完整的天麻苯丙烷类产物合成代谢通路图,并注释了大量相关基因序列。通过本研究为后续深入的基因功能研究及天麻药材品质形成的分子机理研究、天麻栽培过程的质量控制以及采收后的保鲜处理等奠定了基础。
OBJECTIVE: To analyze transcriptomics of Gastrodia elata, and to elucidate the relationship between the content of secondary metabolites and the activity of related enzymes in Gastrodia elata Blume from the transcriptome level. Methods: Illumina Hiseq 4000 was used to analyze the transcriptome of fresh Gastrodia elata and postharvest open 5 days Gastrodia elata, and to annotate and identify differentially expressed genes by comparing with the gene database. RESULTS: Fresh Gastrodia elata and Gastrodia elata Blume 5 days after harvest were obtained 18 772 600, 19 353 329 Clean Reads respectively, and 63 455 Unigene were obtained after assembly by Trinity. Database comparison, and finally get 18 913 annotated Unigene. After 5 days of Gastrodia elata Blume, 953 genes were up-regulated, and 1876 genes were down-regulated. A total of 2 829 differentially expressed genes were found. 634 differentially expressed genes were annotated into 25 functional categories by COG, and 612 differentially expressed genes were annotated by KEGG to 50 metabolic pathways. In the differential expression data of Tianma transcriptome, 11 key genes were annotated by phenylpropanoid pathway in KEGG database. Compared with fresh Gastrodia elata Blume, the expression of CM, PDT, PAL, 4CL, CHS, F5H and F3’M genes of Gastrodia elata Blume Decreased, C4H, CCR expression increased, CAD expression both increased and decreased. Conclusion: In this study, a more complete metabolic pathway of phenylpropane products was obtained, and a large number of related gene sequences were annotated. The study laid the foundation for further research on gene function and molecular mechanism of Gastrodia elata Blume’s quality, quality control of Gastrodia elata cultivation process and preservation after harvest.