基于转录组数据挖掘阴香右旋龙脑生物合成相关基因

Mining Genes Related to Biosynthesis of Cinnamomum burmannii D-borneol Based on Transcriptome Data

右旋龙脑又名天然冰片,是珍贵的传统中药材和高级香料,具有抗菌、消炎、促进药物透过血脑屏障等作用。为阐明右旋龙脑的生物合成途径,收集了2种化学型的阴香,利用Illumina Novaseq测序平台进行转录组测序,使用Trinity软件对Clean reads进行组装,共获得40449条Unigene,N 50长度为1679 bp,Q 30质量值均在92%以上,说明测序质量较好,可用于后续分析。分别有23613(58.38%)、21743(53.75%)、16372(40.48%)、15384(38.03%)、17993(44.48%)、19753(48.83%)个基因在NR、eggNOG、Swiss-Prot、KEGG、Pfam、GO数据库获得注释。KEGG富集分析表明,柠檬烯和蒎烯降解、二苯醚类、二芳庚烷类和姜辣素生物合成被富集,类黄酮生物合成、苯基丙烷生物合成、二萜生物合成等次生代谢物质的合成通路也被富集。GO分析发现,生物学过程类别主要集中在生物过程、转录,DNA模板化;细胞组分类别主要涉及细胞核、细胞质、质膜等细胞组分;分子功能类别主要集中在分子功能、蛋白质结合和序列特异性DNA结合。鉴定出参与右旋龙脑生物合成相关酶基因38个,包括35个参与萜类骨架生物合成有关的酶基因,3个单萜合酶基因,其中MEP途径可能编码5种酶的7个基因和MVA途径可能编码4种酶的7个基因表达量上调。此外与单萜生物合成相关的5个转录因子(bHLH家族2个、MYB家族1个、bZIP家族1个)显著上调。

D-borneol,also known as natural borneol,is a precious traditional Chinese medicine and high-grade spice,which has the functions of anti-bacterial,anti-inflammatory,and promoting drugs to penetrate the blood brain barrier.In order to clarify the biosynthetic pathway of D-borneol,two chemical types of Cinnamomum burmannii were selected,Transcriptome sequening was carried out through Illumina Novaseq sequencing platform,and clean reads were assembled using Trinity software.A total of 40449 genes were obtained,with N 50 length of 1679 bp and Q 30 mass value of more than 92%,which indicated that the sequencing quality was good and could be used for subsequent analysis.23613(58.38%),21743(53.75%),16372(40.48%),15384(38.03%),17993(44.48%)and 19753(48.83%)genes were annotated in NR,eggNOG,Swiss Prot,KEGG,Pfam and GO databases respectively.The KEGG enrichment analysis showed that the degradation of limonene and pinene,the biosynthesis of diphenyl ethers,diarylheptanoids and gingerols were enriched,and the biosynthesis pathways of secondary metabolites such as flavonoids,phenylpropane and diterpenoids were also enriched.GO analysis found that biological processes mainly focused on biological processes,transcription,and DNA templating;The classification of cell component mainly involved cell components such as nucleus,cytoplasm and plasma membrane;molecular functions mainly focused on molecular functions,protein binding and sequence specific DNA binding.38 genes related to the biosynthesis of D-borneol were identified,including 35 genes related to the biosynthesis of terpene skeletons and 3 genes related to monoterpene synthase.Among them,the MEP pathway might encode 7 genes of 5 enzymes and the MVA pathway might encode 7 genes of 4 enzymes,resulting in their gene expression upregulating.In addition,5 transcription factors(2 bHLH,1 MYB,1 bZIP)related to monoterpene biosynthesis were significantly up-regulated.