基于代谢组学的不同品种芍药根系次生代谢产物差异表达分析

Differential Expression Analysis of Secondary Metabolite in Different Varieties of Paeonia lactiflora Based on Metabolomics

以不同芍药品种为试材,采用甲醇漩涡法进行次生代谢产物的提取,超高效液相色谱串联质谱(UPLC-MS/MS)进行成分的分离和鉴定,基于代谢组学进行差异鉴别分析,并基于本地代谢数据库的自建MWDB数据库(Metware Database),对原始数据进行筛选,对不同品种芍药的代谢物进行质控分析、主成分分析、正交偏最小二乘法、聚类分析和样品重复相关性评估,研究了3种芍药品种次生代谢产物之间的差异,以期为进一步对芍药药用价值的研究、栽培育种及遗传改良提供参考依据。结果表明:3个芍药品种中鉴定出萜类、生物碱类、酚酸类、鞣质、木质素和香豆素类共五大类464种次生代谢物;由主成分和偏最小二乘判别分析可知3个芍药品种间的次生代谢物差异明显;差异代谢物的筛选共得到3个品种间显著差异代谢物53种(上调26,下调27);利用KEGG数据库共注释到194条差异代谢途径,富集到次生代谢产物生物合成、苯丙氨酸代谢、烟酸和烟酰胺代谢、糖酵解代谢和苯并恶嗪类生物合成共5条生物合成途径。“红色魅力”“大富贵”“粉玉奴”的次生代谢物的种类和含量存在较大差异,3个芍药品种次生代谢产物差异显著,其中“粉玉奴”适合作为药用品种,“大富贵”适合观赏,3个品种芍药原花青素的差异表达及苯丙氨酸的代谢途径产物可能与3个品种花色的差异有关。

The differential analysis of different P.lactiflora varieties was carried out based on metabolomics.Methanol vortex method was used to extract secondary metabolites,ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was used to separate and identify the components,and original data were screened based on the Metware Database.Quality control analysis,principal component analysis,orthogonal partial least squares discriminant analysis(OPLSDA),cluster analysis and repeated correlation evaluation were carried out for the metabolites of different varieties of peony.The difference of secondary metabolites among three P.lactiflora varieties were studied,in order to provide reference for application,cultivation and genetic improvement of peony.The results showed that 464 kinds of secondary metabolites included terpenoids,alkaloids,phenolic acids,tannins,lignin and coumarins were identified from the three P.lactiflora varieties.Principal component analysis and partial least square discriminant analysis showed that there were significant differences in secondary metabolites among the three peony varieties.A total of 53metabolites with significant difference among 3varieties were obtained(upregulated 26,down-regulated 27).A total of 194differential metabolic pathways were obtained by using KEGG database annotation,in which were enriched to secondary metabolism product biosynthesis pathway,phenylalanine metabolism,niacin and nicotinamide metabolism,glycolytic metabolism and benzoxazine biosynthesis,a total of 5biosynthetic pathways.This study showed that there were significant differences in the types and contents of secondary metabolites among‘Red Charm’‘Dafugui’and‘Fanyunu’,indicated that the secondary metabolites of the three P.lactiflora varieties were significantly different,among which‘Fenyunu’was suitable for medicinal use and‘Dafugui’was suitable for ornamental.The differentially expressed procyanidins and the products of phenylalanine metabolism may be related to the differences in flower color among the three varieties.These results laid a good foundation for further research on the medicinal value,cultivation breeding and genetic improvement of P.lactiflora.