论文部分内容阅读
目的:对乌头的6个器官组织,主根、侧根、茎、叶、花、果实进行转录组测序分析,以研究参与其萜类次级代谢生物合成相关基因的表达谱,挖掘其功能基因。方法:采用Illumina Hi Seq 2500平台测序、组装得unigenes,与公共数据库NR,Swiss-Prot,GO,COG,KOG,KEGG比对、注释以获得差异基因,使用实时荧光定量聚合酶链式反应(Real-time PCR)验证参与萜类次级代谢生物合成的关键基因。结果:本研究共获得156 967 635条Clean Reads(28 254 174 300 bp),经序列合并拼接后获得103 337个unigenes,通过核苷酸和蛋白序列等方面的同源性分析,表明其中37.31%(38 554个unigenes)与其他生物的已知基因具有不同程度的同源性,通过功能分类研究和代谢途径分析(KEGG)获得参与萜类合成158个unigenes(编码5个关键酶)。结论:这些发现的基因将为乌头萜类化合物的生物合成途径及分子机制提供基础数据。
OBJECTIVE: To analyze the transcriptome of 6 organs, main roots, lateral roots, stems, leaves, flowers and fruits of Aconitum to study the expression profiles of the genes involved in the secondary metabolism biosynthesis of terpenoids and to find out their functional genes. Methods: Illumina Hi Seq 2500 platform was used to sequence and assemble unigenes. The sequences were aligned with public databases NR, Swiss-Prot, GO, COG, KOG and KEGG to obtain differential genes. Real-time fluorescence quantitative polymerase chain reaction -time PCR) to validate key genes involved in the secondary metabolism of terpenoids. Results: A total of 156 967 635 Clean Reads (28 254 174 300 bp) were obtained in the present study. 103 337 unigenes were obtained after sequencing and splicing. By homology analysis of nucleotide and protein sequences, it was found that 37.31% (38 554 unigenes) had different degrees of homology with known genes of other organisms. 158 unigenes (encoding 5 key enzymes) involved in terpene synthesis were obtained by functional taxonomy analysis and metabolic pathway analysis (KEGG). Conclusion: These discovered genes will provide basic data for the biosynthesis pathway and molecular mechanism of aconitine.