[关键词]
[摘要]
目的 获得东紫苏Elsholtzia bodinieri转录组数据信息，了解东紫苏挥发油单萜生物合成途径。方法 采用Illumina Hiseq 2000高通量测序获得东紫苏的转录组数据，通过对数据进行组装、拼接及注释，挖掘其单萜类化合物代谢途径相关基因。结果 东紫苏2个样品材料测序后共获得13.67 Gb数据，91 169 356条高质量序列，利用Trinity组装获得93 327条unigenes，平均长度1756 bp。将组装所得到的unigenes分别与NCBI官方的蛋白序列数据库（RefSeq non-redundant proteins，NR）、基因本体论数据库（gene ontology，GO）、真核生物蛋白质同源簇数据库（clusters of orthologous groups for eukaryotic complete genomes，KOG）、京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）等数据库进行BLAST比对分析。通过KEGG代谢通路分析，结果显示有2个单萜代谢相关途径，为萜类骨架生物合成（编号为ko00900）和单萜类生物合成（编号为ko00902），相关unigenes分别有11条和30条；进行实时荧光定量PCR （reverse transcription PCR，RT-PCR）和测序，成功验证6个单萜合成相关候选基因全长unigenes。结论 首次对东紫苏进行高通量转录组测序分析，获得了单萜生物合成的关键酶基因，为后续基因功能的研究奠定基础。
[Key word]
[Abstract]
Objective To obtain the transcriptome data of Elsholtzia bodinieri and understand the pathway of monoterpene biosynthesis of essential oil from E. bodinieri. Methods In this study, Illumina Hiseq 2000 high-throughput sequencing was used to obtain the transcriptome data of E. bodinieri, and the genes related to the metabolism of monoterpenoids were mined by assembling, splicing and annotating the data. Results A total of 13.67 GB data and 91 169 356 clean reads were obtained after sequencing of the two sample materials of E. bodinieri, 93 327 unigenes were obtained by Trinity assembly with an average length of 1756 bp. The unigenes obtained from the assembly were blasted with NR, GO, KOG, KEGG and other databases. KEGG metabolic pathway analysis showed that there were two monoterpene metabolic pathways, terpenoid backbone biosynthesis (No. ko00900) and monoterpenoid biosynthesis (No. ko00902), with 11 and 30 related unigenes, respectively. Full-length unigenes of six monoterpene synthesis related candidate genes were successfully verified by RT-PCR and sequencing. Conclusion For the first time, high-throughput transcriptome sequencing analysis of E. bodinieri was carried out, and the key enzyme genes of monoterpene biosynthesis were obtained, which laid a foundation for the study of subsequent gene functions.
[中图分类号]
R282.12
[基金项目]
国家自然科学基金资助项目（31660082）；云南省科技计划项目[2017FF117（-027）]
