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浙江大学学报（农业与生命科学版）
浙江大学学报（农业与生命科学版）  2023, Vol. 49 Issue (6): 825-839    DOI: 10.3785/j.issn.1008-9209.2022.10.091
食品科学     
基于广泛靶向代谢组学结合高效液相色谱法分析‘紫娟’和‘迎霜’茶树花代谢物差异
朱婉1(),吴颖2(),黎晓湘1,张龙杰3,梁月荣1,陆建良1,郑新强1()
1.浙江大学农业与生物技术学院茶叶研究所, 浙江 杭州 310058
2.宁波市海曙区农业技术管理服务站, 浙江 宁波 315012
3.宁波黄金韵茶业科技有限公司, 浙江 宁波 315412
Analysis of differential metabolites between ‘Zijuan’ and ‘Yingshuang’ tea flowers based on widely targeted metabolomics combined with high performance liquid chromatography
Wan ZHU1(),Ying WU2(),Xiaoxiang LI1,Longjie ZHANG3,Yuerong LIANG1,Jianliang LU1,Xinqiang ZHENG1()
1.Tea Research Institute, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China
2.Agricultural Technology Management and Service Station of Haishu District in Ningbo, Ningbo 315012, Zhejiang, China
3.Ningbo Huangjinyun Tea Science and Technology Co. , Ltd. , Ningbo 315412, Zhejiang, China
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摘要：

为探究茶树品种‘紫娟’和‘迎霜’茶树花之间的风味差异，基于超高效液相色谱-串联质谱法的广泛靶向代谢组学技术，检测2个品种茶树花轻发酵样品中非挥发性代谢物的丰度，并对其进行筛选和鉴定；参照茶叶感官审评方法评价2个样品茶树花各项审评因子，采用高效液相色谱法和比色法检测儿茶素类、黄酮类等滋味成分含量。结果表明：‘紫娟’茶树花滋味甘和较鲜、略苦微涩，‘迎霜’茶树花滋味甘和、微苦略涩。2个品种间有酚酸类（56种）、黄酮类（46种）、脂质（26种）、鞣质（19种）、氨基酸及其衍生物（17种）等219种显著差异代谢物。进一步对其代谢通路进行注释分析发现，氨基酸类物质相关代谢途径以及黄酮和黄酮醇代谢途径是‘紫娟’和‘迎霜’茶树花之间的主要差异代谢途径。此外，‘紫娟’和‘迎霜’茶树花中黄酮类总量、花青素总量、儿茶素类和部分生物碱含量差异显著（P＜0.05），可溶性糖含量差异不显著。上述结果初步说明了黄酮类化合物使茶树花的滋味具有一定苦涩度，相较于‘迎霜’，‘紫娟’茶树花中部分氨基酸及其衍生物含量上调是其茶汤滋味较鲜的主要原因。
关键词： 茶树花;  广泛靶向代谢组学;  ‘紫娟’;  ‘迎霜’;  差异代谢物    
Abstract:

To explore the differences in flavor between flowers of ‘Zijuan’ and ‘Yingshuang’ tea varieties, the abundance of non-volatile metabolites in two light fermented tea flowers was tested, and the metabolites identified via widely targeted metabolomic technology based on ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The evaluation factors were evaluated by tea sensory evaluation methods and the contents of taste components, such as catechins and flavonoids, were detected by high performance liquid chromatography and colorimetric method. The results showed that ‘Zijuan’ tea flowers tasted sweet and fresh, and a little bitter and slightly astringent, while ‘Yingshuang’ tea flowers tasted sweet, slightly bitter and a little astringent. There were 219 kinds of significantly differential metabolites, including phenolic acids (56 kinds), flavonoids (46 kinds), lipids (26 kinds), tannins (19 kinds), amino acids and their derivatives (17 kinds) and others between the two varieties. Furthermore, metabolic pathway annotations revealed that amino acid-related metabolic pathways and flavone and flavonol metabolism pathways were the main differential metabolic pathways between ‘Zijuan’ and ‘Yingshuang’ tea flowers. In addition, the total contents of flavonoids and anthocyanins and the contents of catechins and some alkaloids between ‘Zijuan’ and ‘Yingshuang’ tea flowers were significantly different (P＜0.05), while the differences in the soluble sugar contents were not significant. The above results preliminarily showed that the flavonoids made the taste of tea flowers had a certain degree of bitterness and astringency, while compared with those in ‘Yingshuang’, the increase of contents of some amino acids and their derivatives in ‘Zijuan’ tea flowers was the main reason for its fresh taste.
Key words: tea flowers    widely targeted metabolomics    ‘Zijuan’    ‘Yingshuang’    differential metabolites
收稿日期: 2022-10-09 出版日期: 2023-12-25
CLC:  TS272.5  
基金资助: 浙江省宁波市“科技创新2025”重大专项(2019B10022);国家现代农业产业技术体系建设专项(CARS-19)
通讯作者: 郑新强     E-mail: 22016175@zju.edu.cn;76460140@qq.com;xqzheng@zju.edu.cn
作者简介: 朱婉（https://orcid.org/0000-0003-2557-4724），E-mail：22016175@zju.edu.cn|吴颖（https//:orcid.org/0000-0003-2160-3946），E-mail：76460140@qq.com
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朱婉
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引用本文:

朱婉,吴颖,黎晓湘,张龙杰,梁月荣,陆建良,郑新强. 基于广泛靶向代谢组学结合高效液相色谱法分析‘紫娟’和‘迎霜’茶树花代谢物差异[J]. 浙江大学学报（农业与生命科学版）, 2023, 49(6): 825-839.

Wan ZHU,Ying WU,Xiaoxiang LI,Longjie ZHANG,Yuerong LIANG,Jianliang LU,Xinqiang ZHENG. Analysis of differential metabolites between ‘Zijuan’ and ‘Yingshuang’ tea flowers based on widely targeted metabolomics combined with high performance liquid chromatography. Journal of Zhejiang University (Agriculture and Life Sciences), 2023, 49(6): 825-839.

链接本文:

https://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2022.10.091    或    https://www.zjujournals.com/agr/CN/Y2023/V49/I6/825

样品

Sample

外形

Appearance

叶底

Infused leaves

汤色

Infusion color

香气

Aroma

滋味

Taste

ZJF

花朵完整，花瓣浅黄带浅绿，花托微黄，

花萼绿翠，较匀整

成朵透亮、花蕊完整，

花萼边缘略有红边

黄明亮蜜香稍闷甘和较鲜、略苦微涩
YSF

花朵完整，花瓣象牙白，花托稍带嫩绿，

花萼深绿，较匀整

成朵透亮、花蕊完整杏黄明亮略有花香甘和、微苦略涩
表1  ZJF和YSF感官审评结果
图1  质控样本的TIC重叠图A.正离子模式；B.负离子模式。

指标

Index

PC1PC2PC3PC4PC5

特征值

Eigenvalue

613.0989.6571.0366.7056.54

累积贡献率

Cumulative contribution

rate/%

68.3578.3486.2693.70100.00

方差占比

Proportion of variance/%

68.359.997.927.446.30
表2  ZJF和YSF代谢物的PCA结果
图2  ZJF和YSF中鉴定到的代谢物类别括号中数字为该类代谢物数量占所鉴定到的897种代谢物的百分比。
图3  ZJF和YSF代谢物的多元统计分析结果A. PCA得分图；B. OPLS-DA模型图。
图4  ZJF和YSF的显著差异代谢物图谱A.显著差异代谢物聚类树；B.显著差异代谢物火山图。图例中颜色标尺为代谢物响应值（相对含量）进行－1.5～1.5标准化处理后得到的数值（红色表示代谢物相对含量上调，绿色表示代谢物相对含量下调）。
图5  ZJF和YSF中219种显著差异代谢物饼图括号中数字为该类差异代谢物数量占ZJF和YSF中219种显著差异代谢物的百分比。
图6  ZJF和YSF中显著差异代谢物的KEGG通路富集图富集因子表示差异表达的代谢物中在对应通路中的个数与该通路中注释到的代谢物总数的比值。
  
  
图7  YSF和ZJF中的主要滋味差异代谢物A.黄酮类；B.氨基酸及其衍生物；C.酚酸类；D.鞣质。图例中颜色标尺为对代谢物响应值（相对含量）进行0～1标准化处理后得到的数值。
化学组分 Chemical componentZJFYSF
GC/(mg/g)1.54±0.16b2.52±0.10a
EGC/(mg/g)13.92±0.33a11.10±0.36b
C/(mg/g)1.10±0.13a0.77±0.05b
EC/(mg/g)8.83±0.21a4.57±0.04b
非酯型儿茶素 Non-ester catechins/(mg/g)25.39±0.30a18.96±0.20b
EGCG/(mg/g)17.93±1.02b39.22±2.35a
GCG/(mg/g)0.44±0.02b0.75±0.06a
ECG/(mg/g)10.91±0.40b19.70±1.22a
CG/(mg/g)0.24±0.03a0.20±0.03a
酯型儿茶素 Ester catechins/(mg/g)29.52±0.76b59.87±3.65a
酯型儿茶素/非酯型儿茶素比值 Ratio of ester catechins to non-ester catechins1.16±0.02b3.16±0.16a
可可碱 Theobromine/(mg/g)0.16±0.00b0.28±0.01a
茶碱 Theophylline/(mg/g)0.21±0.00a0.18±0.01b
咖啡碱 Caffeine/(mg/g)8.88±0.19a8.63±0.68a
黄酮类 Flavonoids/(mg/g)111.89±7.39a79.90±2.43b
可溶性糖 Soluble sugar/(mg/g)264.36±16.86a274.22±4.43a
花青素 Anthocyanin/(μmol/g)152.51±6.38a132.64±1.52b
表3  ZJF和YSF中的滋味化学组分含量
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