LI Yonghui,GUO Xiangmeng,GAO Fei,et al.Differential Expression Analysis of Secondary Metabolite in Different Varieties of Paeonia lactiflora Based on Metabolomics[J].Northern Horticulture,2023,(08):46-55.[doi:10.11937/bfyy.20223284]
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
- Title:
- Differential Expression Analysis of Secondary Metabolite in Different Varieties of Paeonia lactiflora Based on Metabolomics
- 关键词:
- 芍药; 多元统计分析; 超高液相色谱串联质谱; 代谢组学; 差异表达
- Keywords:
- Paeonia lactiflora Pall; multivariate statistical analysis; UPLC-MS/MS; metabolomics; differential expression
- 文献标志码:
- A
- 摘要:
- 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。
- Abstract:
- 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 (OPLS-DA),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 53 metabolites with significant difference among 3 varieties were obtained (up-regulated 26,down-regulated 27).A total of 194 differential 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 5 biosynthetic 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.
参考文献/References:
[1]MAO Q Q,IP S P,TSAI S H,et al.Antidepressant-like effect of peony glycosides in mice[J].Journal of Ethnopharma,2008,119:272-275.[2]AHN M S,PARK P H,KWON Y N,et al.Discrimination of floral scents and metabolites in cut flowers of Peony (Paeonia lactiflora Pall.) cultivars[J].Korean Journal of Plant Resources,2018,31(6):641-651.[3]ZHENG Q W,JIANG X,SUN T,et al.Total glucosides of paeony for the treatment of psoriasis:A systematic review and meta-analysis of randomized controlled trials[J].Phytomedicine,2019,62:152940.[4]吴普.神农本草经(孙星衍,孙冯骥,辑)[M].北京:人民卫生出版社,1982.[5]ZHAI A,ZHANG Z,KONG X.Paeoniflorin alleviates H2O2-induced oxidative injury through down-regulation of MicroRNA-135a in HT-22 ells[J].Neurochemical Research,2019,44(12):2821-2831.[6]国家药典委员会.中华人民共和国药典:一部[S].北京:中国医药科技出版社,2015.[7]KIM S A,JANG E S,LEE A Y.Anti-inflammatory and anti-oxidant effects of oxypaeoniflorin,paeonifloriand Paeonia lactiflora cv.‘Red Charm’ flower petal extracts in macrophage cells[J].Korean Journal of Plant Resources,2020,33(3):153-162.[8]DEVAUX P G,HORNINA M G,HORNING E C.Benyzl-oxime derivative of steroids:A new metabolic profile procedure for human urinary steroids[J].Analytical Letters,1971,4(3):151-160.[9]NICHOLSON J K,LINDON J C,HOLMES E.‘Metabonomics’:Understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data[J].Xenobiotica;the Fate of Foreign Compounds in Biological Systems,1999,29(11):1181-1189.[10]朱超,梁琼麟,王义明,等.代谢组学的整合化发展及其新进展[J].分析化学,2010,38(7):1060-1068.[11]饶怡菲,江志辉,何明珍,等.基于超高效液相色谱-串联质谱法研究宽缨酮在大鼠体内的生物利用度及代谢产物[J].中药新药与临床药理,2020,31(4):455-463.[12]LIU B,SONG Z,YU J,et al.The atherosclerosis-ameliorating effects and molecular mechanisms of Bu Yang Huan Wu decoction[J].Biomed Pharmacother,2020,123:109664.[13]ZHANG L,WEI W.Anti-inflammatory and immunoregulatory effects of paeoniflorin and total glucosides of paeony[J].Pharmacology and Therapeutics,2020,207:107452.[14]ZHAI A,ZHANG Z,KONG X J.Paeoniflorin alleviates H2O2-induced oxidative injury through down-regulation of microRNA-135a in HT-22 cells[J].Neurochem Research,2019,44(12):2821-2831.[15]彭国梅,张启云,李冰涛,等.UPLC-MS/MS 同时测定葛根芩连汤含药血清10个有效成分的含量[J].中药新药与临床药理,2016,27(3):393-398.[16]闫寒,付衍,彭娟,等.大鼠含药血清中芍药苷的含量测定[J].中国实验方剂学杂志,2013,19(14):89-91.[17]宋祯彦,何攀,李富周,等.高效液相色谱串联质谱法测定当归芍药散提取物和含药血清中 3 种活性成分含量[J].中国中医药信息杂志,2021,28(1):93-99.[18]付新,张洪财,张贺,等.HPLC 测定当归芍药散中的芍药苷和芍药内酯苷的含量[J].化学工程师,2020,34(1):25-27.[19]贺晓艳,宋秋月,郑丹,等.HPLC 法测定当归芍药散中芍药苷、白芍苷、阿魏酸、洋川芎内酯I的含量[J].亚太传统医药,2011,7(7):36-39.[20]JIE W J,LOU H Y,LIU Y c,et al.Profiling alkaloids in Aconitum pendulum N.Busch collected from different elevations of Qinghai province using widely targeted metabolomics[J].Phytochemistry,2022(195):113047.[21]THVENOT E A,ROUX A,XU Y,et al.Analysis of the human adult urinary metabolome variations with age,body mass index,and gender by implementing a comprehensive workflow for univariate and OPLS statistical analyses[J].Journal of Proteome Research,2015,14(8):3322-3335.[22]KANEHISA M,GOTO S.KEGG:Kyoto encyclopedia of genes and genomes[J].Nucleic Acids Research,2000,28(1):27-30.[23]时羽杰,邬晓勇,糜加轩,等.核桃内种皮苦涩味品质代谢组学分析[J].西北农林科技大学学报(自然科学版),2021,49(6):55-64.[24]司仕英,孟秦语,孙道阳,等.关中地区引种芍药品种的综合性状评价[J].西北林学院学报,2021,36(6):134-139.[25]李健,韩晨静,王琦,等.温湿度调控对芍药花期的影响[J].山东农业科学,2021,53(12):74-77.[26]AKOWUAH G,SADIKUN A,AHMAD M.Flavonoid identification and hypoglycaemic studies of the butanol fraction from Gynuraprocumbens[J].Pharm Biology,2008,40(6):405-410.[27]康晓飞.芍药根内主要活性物质与营养成分的研究[D].泰安:山东农业大学,2011.[28]方贤胜,吴涛,肖良俊.基于广泛靶向代谢组学的浅黄色和紫色核桃内种皮成分差异分析[J].食品科学,2021,42(12):215-221.[29]金英善,陈曼丽,金银哲,等.芍药花活性成分分析及体外清除自由基活性研究[J].扬州大学学报(农业与生命科学版).2012,33(3):86-90.[30]陈东方.复配芍药提取物在抗衰老药妆产品中的研究[D].上海:上海交通大学,2016.[31]刘雨艳.牡丹中芍药苷降血脂活性的研究[J].石油石化物资采购,2020(27):152-153.[32]熊欢欢,张含国,张磊,等.干旱胁迫下长白落叶松的代谢组学分析[J].东北林业大学学报,2019,47(4):1-7.[33]DOUGLAS C J.Phenylpropanoid metabolism and lignin biosynthesis:From weeds to trees[J].Trends in Plant Science,1996,1(6):171-178.[34]MIYAMOTO K,FUJITA M,SHENTON M R,et al.Evolutionary trajectory of phytoalexin biosynthetic gene clusters in rice[J].The Plant Journal:For Cell and Molecular Biolog,2016,87(3):293-304.[35]欧阳光察,薛应龙.植物苯丙烷类代谢的生理意义及其调控[J].植物生理学通讯,1988(3):9-16.[36]POURCEL L,ROUTABOUL J M,KERHOA S L,et al.TRANSPARENT TESTA10 encodes a laccase-like enzyme involved in oxidative polymerization of flavonoids in Arabidopsis seed coat[J].Plant Cell,2005,17(11):2966-2980.[37]MARLES M A S,VANDENBERG A,BETT K E.Polyphenol oxidase activity and differential accumulation of polyphenolics in seed coats of pinto bean (Phaseolus vulgaris L.) characterize postharvest color changes[J].Journal of Agricultural and Food Chemistry,2008,56(16):7049-7056.
相似文献/References:
[1]刘 萍,邵晓琪,丁义峰,等.不同芍药品种花瓣中营养成分含量的比较[J].北方园艺,2014,38(01):151.
LIU Ping,SHAO Xiao-qi,DING Yi-feng,et al.Comparison on Nutritional Components Contents in Petals of Six Peony Varieties[J].Northern Horticulture,2014,38(08):151.
[2]孙晓梅,王慧聪,周文强,等.芍药胚离体培养的初步研究[J].北方园艺,2013,37(10):97.
SUN Xiao-mei,WANG Hui-cong,ZHOU Wen-qiang,et al.Preliminary Study on Embryo Culture in vitro of Paeonia lactiflora Pall.[J].Northern Horticulture,2013,37(08):97.
[3]薛银芳,赵大球,周春华,等.芍药组织培养的研究进展[J].北方园艺,2012,36(04):167.
XUE Yin-fang,ZHAO Da-qiu,ZHOU Chun-hua,et al.Research Advances of Herbaceous Peony Tissue Culture[J].Northern Horticulture,2012,36(08):167.
[4]孙晓梅,张萌萌,高昊丹,等.芍药种皮特性的研究[J].北方园艺,2012,36(06):55.
SUN Xiao-mei,ZHANG Meng-meng,GAO Hao-dan,et al.Study on Characteristic for Seed Coat of Paeonia lactiflora[J].Northern Horticulture,2012,36(08):55.
[5]刘萍,王贺贺,丁义峰,等.维生素C与甘氨酸甜菜碱对芍药花瓣衰老生理的调节作用[J].北方园艺,2012,36(06):158.
LIU Ping,WANG He-he,DING Yi-feng,et al.Regulation of VC and GB on Senescence Physiology in Herbaceous Peony Petals[J].Northern Horticulture,2012,36(08):158.
[6]孙燕,张福海,李成忠,等.不同供硅水平对芍药光合特性及水分利用效率的影响[J].北方园艺,2015,39(08):81.[doi:10.11937/bfyy.201508023]
SUN Yan,ZHANG Fu-hai,LI Cheng-zhong,et al.Effect of Different Levels of Silicon Supply on the Photosynthetic Properties and Water Utilization Efficiency of Paeonia lactiflora Pall.[J].Northern Horticulture,2015,39(08):81.[doi:10.11937/bfyy.201508023]
[7]孙晓梅,王思思,杨宏光,等.芍药种子萌发过程中抗氧化酶活性变化规律的研究[J].北方园艺,2015,39(14):53.[doi:10.11937/bfyy.201514016]
SUN Xiaomei,WANG Sisi,YANG Hongguang,et al.Antioxidant Enzymes Activity of Paeonia lactiflora During Seed Germination[J].Northern Horticulture,2015,39(08):53.[doi:10.11937/bfyy.201514016]
[8]高 昌 勇.芍 药 胚 离 体 培 养 的 研 究[J].北方园艺,2010,34(16):0.[doi:10.11937/bfyy.201016053]
GAO Chang -yong.Study on Embryo Culture of Chinese Herbaceous Peony in vitro[J].Northern Horticulture,2010,34(08):0.[doi:10.11937/bfyy.201016053]
[9]牛 立 军,符 小 宁,刘 爱 青.观赏芍药在花境中的应用[J].北方园艺,2010,34(04):0.[doi:10.11937/bfyy.201004039]
NIU Li - jun,FU Xiao - ning,LIU Ai - qing.Application of Herbaceous Peony in Flower Border[J].Northern Horticulture,2010,34(08):0.[doi:10.11937/bfyy.201004039]
[10]孙晓梅,崔金秋,王慧聪,等.芍药属植物杂交亲和性及杂交后代ISSR分子鉴定[J].北方园艺,2016,40(09):98.[doi:10.11937/bfyy.201609026]
SUN Xiaomei,CUI Jinqiu,WANG Huicong,et al.Hybridization Affinity of Paeonia Plant and ISSR Molecular Identification of Hybrids[J].Northern Horticulture,2016,40(08):98.[doi:10.11937/bfyy.201609026]
备注/Memo
第一作者简介:李勇慧(1977-),女,硕士,副教授,现主要从事植物学药用成分分析等研究工作。E-mail:huiyongli8209 @126.com.基金项目:国家自然科学基金面上资助项目(31870697);河南省自然科学基金资助项目(202300410287)。收稿日期:2022-08-10