丹参酮生物合成途径CYP450基因相关长链非编码RNA的筛选

Screening of long non-coding RNA related to CYP450s involved in biosynthesis of tanshinones

丹参酮是中药丹参的主要有效成分,其生物合成途径表明细胞色素CYP450酶在丹参酮生物合成途径结构后修饰过程中发挥着重要作用。长链非编码RNA(lncRNA)是一类长度大于200 nt的核苷酸,对药用植物的生长发育、次生代谢等具有重要调控作用。该研究通过对丹参毛状根进行诱导,利用高通量测序构建其长链非编码RNA文库,共获得8 942个差异lncRNA,其中6 755个基因间lncRNA,并鉴定出1 115 814个lncRNA-靶基因对,包括122个lncRNA-靶基因顺式对。对差异lncRNA与CYP450进行相关性分析,共鉴定出16 249个lncRNA-CYP450靶基因对。将其与丹参酮生物合成途径CYP76AH1,CYP76AH3,CYP76AK1基因进行靶向相关性分析,得到216个靶基因,这些候选基因为丹参酮生物合成途径下游调控机制研究奠定基础。

Tanshinones are abietane-type norditerpenoid quinones that make up the main bioactive ingredients of traditional Chinese medicine Salvia miltiorrhiza.Cytochrome CYP450 plays an important role in the post-structural modification of tanshinone biosynthesis pathway.Long non-coding RNA(lncRNA)have been defined as transcripts longer than 200 nucleotides,which have been functionally characterized in regulating the growth and development,secondary metabolism and stress of medicinal plants.In this study,we perform a comprehensive identification of lncRNAs in response to tanshinone metabolism induced by yeast extract(YE)and Ag^+S.miltiorrhiza hairy roots.Deep RNA sequencing was used to identify a set of different 8 942 lncRNAs,of which 6 755 were intergenic lncRNAs.We predicted a total of 1 115 814 lncRNA-coding gene pairs,including 122 lncRNA-coding gene as cis pairs.The correlation analysis between lncRNA and CYP450 related to tanshinone biosynthesis was carried out and a total of 16 249 lncRNA-CYP450 target gene pairs were identified.Further analysis with functional known CYP76 AH1,CYP76 AH3 and CYP76 AK1 involved in tanshinone biosynthesis,we also identified a set of 216 target genes.These candidate genes will be the important target in the downstream regulation mechanism analysis of the tanshinone biosynthesis pathway.