耳叶牛皮消的全长转录组测序及其重要活性物质代谢通路的分析

Full-length Transcriptomic Sequencing of Cynanchum auriculatum and Metabolic Pathway Analysis of Important Active Substances

耳叶牛皮消(Cynanchum auriculatum)为萝藦科(Asclepiadaceae)植物,具有较高的营养价值和药用潜力,是中国传统的药食同源兼用植物。当前耳叶牛皮消的相关研究以活性物质的提取和分类为主,但分子水平的研究较少。本研究借助PacBio三代测序平台,对‘滨乌1号’耳叶牛皮消的转录组进行测序,获得高质量Unigenes共42710条,总长度为113782167 bp。将上述Unigenes与Nr、KEGG、GO、COG、Swissprot、Pfam等数据库比对,分别获得41642、19502、21997、31150、37710、37433条注释,其中在“氨基酸代谢”(1967条)、“脂质代谢”(1503条)、“萜类化合物和聚酮化合物代谢”(350条)等通路的注释有助于揭示耳叶牛皮消的药理物质代谢机制。进一步分析表明,类黄酮合成通路上可注释到29条Unigenes,黄酮和黄酮醇合成通路上可注释到2条Unigenes;类固醇合成通路上可注释到68条Unigenes,油菜素类固醇合成通路上可注释到12条Unigenes。通过这些潜力基因的鉴定,可以为耳叶牛皮消活性物质代谢的分子机制研究提供科学依据。此外,共检索到2325个SSR位点和517个lncRNA,为耳叶牛皮消的种质鉴定、遗传图谱构建提供基础。同时,挖掘出42个转录因子,以bHLH家族、ERF家族和TCP家族为主,它们可能参与了耳叶牛皮消的次级代谢产物调控网络。本研究为耳叶牛皮消的全长转录组进行注释,为耳叶牛皮消的重要活性物质代谢机制和功能基因研究奠定了理论基础。

Cynanchum auriculatum is a plant of Asclepiadaceae family,with high nutritional value and medicinal potential,and is a traditional homology of medicine and food in China.At present,C.auriculatum-related studies focused on the extraction and classification of active substances,but only a few focused on the molecular level.Through the third-generation sequencing platform of PacBio,the full-length transcriptome of‘Binwu No.1’C.auriculatum was sequenced.A total of 42710 high-quality Unigenes with a total length of 113782167 bp were obtained.By comparing these Unigenes with Nr,KEGG,GO,COG,Swissprot and Pfam databases,41642,19502,21997,31150,37710 and 37433 annotations were obtained respectively.Among them,the annotation in the pathways,such as“Amino Acid Metabolism”(1967),“Lipid Metabolism”(1503),“Terpenoids and Polyketide Metabolism”(350),was helpful to reveal the metabolic mechanism of pharmacological substances in C.auriculatum.Further analysis showed that,29 Unigenes were annotated on the flavonoid synthesis pathway,2 Unigenes were annotated on the flavone and flavonol synthesis pathway,and there were 68 Unigenes on the steroid synthesis pathway and 12 Unigenes on the brassinosteroid synthesis pathway.The identification of these potential genes can provide a scientific basis for the molecular mechanism study of the active substances metabolism in C.auriculatum.In addition,a total of 2325 SSR loci and 517 lnc-RNAs were retrieved,which could provide a basis for germplasm identification and genetic map construction of C.auriculatum.At the same time,42 transcription factors were unearthed,mainly bHLH family,ERF family and TCP family,which may be involved in the secondary metabolite regulatory network of C.auriculatum.This research is the first to annotate the full-length transcriptome of C.auriculatum,laying a theoretical foundation for the study of important active substances metabolism mechanisms and functional genes of C.auriculatum.