虎尾轮转录组测序及生物信息学分析

Sequencing and Bioinformatics Analysis of Uraria crinita(L.)Desv.ex DC

虎尾轮(Uraria crinita(L.)Desv.ex DC)为闽南民间常用保肝护肝中草药,是一种极有使用、研究、开发价值的天然药用植物资源,但对其分子机制研究严重滞后,在一定程度上限制了该民间资源的利用和开发。本实验通过Illumina Hiseq对虎尾轮的根和茎转录组测序,采用Trinity软件完成虎尾轮高质量数据转录本拼装,而后进行生物信息学分析。研究结果表明:总共获得的高质量unigene有255407896个,N50的长度是992 bp,其平均长度是559.07 bp;将虎尾轮的122207条unigene与NR、GO、KEGG等7个重要的数据库系统进行比对,获得了虎尾轮基因功能的注释信息,结果表明,有75530条unigene至少注释到1个数据库,占61.8%;虎尾轮的根对茎的差异表达unigene有19714个,其中有9038个上调表达,10676个下调表达。在对虎尾轮根与茎差异基因分析中,利用KEGG数据库代谢途径对132个显著性KO途径分析,发现共挖掘出114个unigene与虎尾轮黄酮类生物合成途径密切相关。提取虎尾轮转录组中与黄酮类生物合成密切相关基因的TPM值,60.71%的基因在根中的TPM值高于茎中的TPM植,根中的表达远高于茎的表达,符合民间习用其根的用法。以上结果为今后进一步挖掘虎尾轮药用活性成分的功能基因、生物合成调控、品质改良等提供参考依据。

Uraria crinita(L.)Desv.ex DC is a commonly used Chinese herbal medicine for protecting the liver and protecting the liver in southern Fujian.It is a natural medicinal plant resource with great use,research and development value.However,the research on its molecular mechanism is seriously lagging behind,which limits the utilization and development of this folk resource to a certain extent.In this experiment,Illumina Hiseq was used to sequence the root and stem transcripts of Uraria crinita(L.)Desv.ex DC,complete the assembly of high-quality data transcripts of Uraria crinita(L.)Desv.ex DC with Trinity software,and then bioinformatics analysis was carried out.The research results showed that 255407896 high-quality unigenes were obtained,with an N50 length of 992 bp and an average length of 559.07 bp.122207 unigenes of Uraria crinita(L.)Desv.ex DC were compared with NR,GO,KEGG,etc,and their function annotation information was obtained.The results showed that 75530 unigenes were annotated to at least one database,accounting for 61.8%.There were 19714 root to stem differential expressions of unigene in Uraria crinita(L.)Desv.ex DC,of which 9038 were up-regulated and 10676 were down-regulated.In the analysis of different genes in the root and stem of Uraria crinita(L.)Desv.ex DC,through the analysis of metabolic pathway in KEGG database and the further analysis of 132 significant KO pathways,it was found that a total of 114 unigenes were closely related to the flavonoid biosynthesis pathway of Uraria crinita(L.)Desv.ex DC.The TPM value of the genes closely related to flavonoids biosynthesis in the transcription group of Uraria crinita(L.)Desv.ex DC was extracted.The TPM value of 60.71% of the genes in the root was higher than that in the stem,and the expression in the root was much higher than that in the stem,which was consistent with the folk usage of its root.The above results provide a reference for further exploring the functional genes,biosynthesis regulation and quality improvement of the active ingredients of Uraria crinita(L.)Desv.ex DC in the future.