目的 对八爪金龙根进行转录组测序，挖掘次生代谢合成相关基因，探索八爪金龙次生代谢产物生物合成的分子基础。方法 采用Illumina HiSeq 4000高通量测序技术，对八爪金龙根进行转录组测序。使用Trinity软件对获得的Unigenes数据进行过滤组装，运用KEGG数据库对Unigenes进行注释。结果 共获得52 249条Unigenes，其中31 391条被公共数据库成功注释，1507条Unigenes被注释到次生代谢产物合成。通过对转录组数据深入挖掘发现，参与八爪金龙苯丙素生物合成的Unigenes共有126条，参与萜类化合物骨架生物合成的Unigenes数量为73条，参与黄酮类化合物生物合成Unigenes有58条，参与次生代谢后修饰的Unigenes共有253条。筛选出9条Unigenes编码4个与八爪金龙香豆素生物合成的关键酶，23条Unigenes编码8个与黄酮生物合成的关键酶，39条Unigenes编码19个与萜类生物合成的关键酶，140条CYP450基因和113条UGT基因可能参与次生代谢物的修饰。微卫星识别软件（microsatellite identification tool，MISA）分析发现八爪金龙转录组包含17 400个简单重复序列（simple sequence repeats，SSR）。结论 通过高通量转录组测序，初步揭示了参与八爪金龙次生代谢产物合成相关的基因，为进一步研究八爪金龙次生代谢产物合成途径关键酶的功能及其调控机制奠定了基础。

Objective To explore related genes of the secondary metabolic biosynthesis and their molecular basis of Ardisia crispa root by using high-throughput sequencing. Methods The high-throughput RNA-seq technology (Illumina HiSeq 4000) was used to conduct a transcriptomic analysis of A. crispa root. Unigenes splicing was performed using Trinity software and annotated with KEGG database to predict the specific pathways. Results A total of 52 249 Unigenes were obtained, of which 31 391 were annotated in the public database, 1507 Unigenes were annotated for pathways of secondary metabolism biosynthesis. Based on the bioinformatic analysis, 126 Unigenes were involved in the phenylpropanoid biosynthesis, 73 Unigenes might participate in terpenoid backbone biosynthesis, 58 Unigenes were involved in the flavonoid biosynthesis, and 253 Unigenes might participate in secondary metabolism post-modification. A total of 9, 23, 39 Unigenes were screened out from the transcriptome database, which encoded 4, 8, 19 key metabolic enzymes associated with coumarin, flavone and terpenoid biosynthesis, respectively. 140 CYP450 genes and 113 UGT genes may be involved in the modification of secondary metabolites. MISA prediction found 17 400 simple sequence repeats (SSRs). Conclusion The high-throughput transcriptome sequencing revealed the genes involved in the synthesis of flavonoids and triterpenes, which lays a foundation for further studies on function and regulatory mechanisms of key enzymes involved in the synthesis of secondary metabolites in A. crispa.

R282.12

国家重点研发计划项目（2018YFC1708100）；贵州省高层次创新型人才培养项目（黔科合人才[2015]4032号）；贵州省科技厅学术新苗项目（黔科合平台人才[2018]5766号-9）；贵州中医药大学博士启动基金（贵中医博士启动（[2019]04号）