防风是一种常用的散装药用植物。探讨色酮和香豆素生物合成的关键酶基因及其表达,关键的活动组件,我们对1岁(S1)和2岁(S2)的S.divaricata植物进行了转录组测序(IlluminaHiSeq)和生物信息学分析。总共获得了40.8Gb的数据。在通过三位一体进行序列组装之后,获得110732个转录本和86233个单基因,与NR对齐并注释,Swiss-Prot,GO,KEGG,PFAM。Daucuscarota和S.divaricata具有最高的序列同源性。KEGG途径富集表明,差异表达基因主要富集在植物激素信号转导中,苯丙素生物合成,和类黄酮生物合成途径。共有27个差异表达的单基因,包括13个酶基因,在与八七中活性成分合成相关的途径中被鉴定。与S1植物相比,S2植物显示PAL的表达上调,BGL,C4H,4CL,CYP98A,CSE,REF,和CCoAOMT以及CHS的表达下调,CAD,和COMT。HCT和POD具有上调和下调的单基因。其中,PAL,C4H,4CL,BGL,CHS和CHS可以用作合成散斑S中活性成分的候选基因。通过RT-qPCR验证了4个关键酶基因,结果与转录组测序结果一致。本研究丰富了双歧杆菌的遗传信息,为次生代谢产物生物合成中候选基因的鉴定提供了支持。
Saposhnikovia divaricata is a commonly used bulk medicinal plant. To explore the key enzyme genes and their expression in the biosynthesis of chromone and coumarin, the key active components, we carried out transcriptome sequencing(Illumina HiSeq) and bioinformatics analysis for the 1-year-old(S1) and 2-year-old(S2) plants of S. divaricata. A total of 40.8 Gb data was obtained. After the sequence assembly via Trinity, 110 732 transcripts and 86 233 unigenes were obtained, which were aligned and annotated with NR, Swiss-Prot, GO, KEGG, and PFAM. Daucus carota and S. divaricata had the highest sequence homology. KEGG pathway enrichment showed that the differentially expressed genes were mainly enriched in plant hormone signal transduction, phenylpropanoid biosynthesis, and flavonoid biosynthesis pathways. A total of 27 differentially expressed unigenes, including 13 enzyme genes, were identified in the pathways related to the synthesis of active ingredients in S. divaricata. Compared with S1 plant, S2 plant showed up-regulated expression of PAL, BGL, C4H, 4CL, CYP98A, CSE, REF, and CCoAOMT and down-regulated expression of CHS, CAD, and COMT. HCT and POD had both up-regulated and down-regulated unigenes. Among them, PAL, C4H, 4CL, BGL, and CHS can be used as candidate genes for the synthesis of the active ingredients in S. divaricata. The four key enzyme genes were verified by RT-qPCR, which showed the results consistent with transcriptome sequencing. This study enriches the genetic information of S. divaricata and provides support for the identification of candidate genes in the biosynthesis of secondary metabolites.