基于非靶向代谢组学研究做青工序对肉桂茶代谢产物的影响

林馥茗; 孙威江

doi:10.19303/j.issn.1008-0384.2022.007.013

基于非靶向代谢组学研究做青工序对肉桂茶代谢产物的影响

doi: 10.19303/j.issn.1008-0384.2022.007.013

林馥茗^{1, 2,},
孙威江^{1, 3, ,}

1.
福建农林大学园艺学院，福建　福州　350002
2.
福建农林大学安溪茶学院，福建　泉州　362000
3.
福建省茶产业工程技术研究中心，福建　福州　350002

基金项目: 福建省教育厅中青年教师教育科研项目（JT180117）；安溪茶学院青年骨干培养基金项目（ACKY2015004)

详细信息

作者简介:
林馥茗（1986−），女，硕士，讲师，研究方向：茶叶加工与品质化学（E-mail：497147822@qq.com）

通讯作者:
孙威江（1964−），男，博士，教授，研究方向：茶叶资源育种与标准化（E-mail：swj8103@126.com）

中图分类号: S 609
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出版历程
- 收稿日期: 2021-12-29
- 录用日期: 2021-12-29
- 修回日期: 2022-04-20
- 网络出版日期: 2022-08-07
- 刊出日期: 2022-07-28

Effect of Rocking-withering on Metabolites in Wuyi Rougui Tea Leaf

LIN Fuming^{1, 2
,},
SUN Weijiang^{1, 3
, ,}

1.
College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
2.
Anxi College of Tea Science, Fujian Agriculture and Forestry University, Quanzhou, Fujian　362000, China
3.
Tea Industry Engineering Technology Research Center of Fujian Province, Fuzhou, Fujian　350002, China

摘要

摘要: 目的通过GC-TOF-MS非靶向代谢组学分析茶叶做青工序对武夷肉桂茶树叶片代谢物的影响，探究做青对乌龙茶品质形成的作用机制，并提供理论参考。方法采摘武夷肉桂中开面3、4叶，经短时间晒青后，按照闽北乌龙茶的做青工艺制成做青组样品，将叶片自然摊放作为对照组（CK），采用气相色谱−飞行时间质谱检测样品，运用多元统计分析OPLS-DA 和单变量统计分析相结合的方法筛选差异代谢物，并进行代谢通路KEGG富集分析。结果与晒青叶相比，对照组及做青组样品中能够鉴定出的差异代谢物分别为21个和14个，其含量主要呈现上调趋势；与对照组相比，做青组样品中显著上调和下调的差异代谢物分别为9个和3个；检测到的差异代谢物包括莽草酸、柠檬酸、L-苹果酸、棉子糖、麦芽糖等有机酸和糖类物质。结论做青工序显著刺激了三羧酸循环中部分中间产物以及莽草酸途径中莽草酸含量发生变化，这主要与氨基酸、碳水化合物等物质代谢有关；其中莽草酸含量在做青结束时显著降低，而苯丙氨酸含量有所提高，这可能与茶叶芳香族化合物的合成有关。
- 武夷肉桂 /
- 做青 /
- 非靶向代谢组学 /
- 初级代谢物
Abstract: Objective Effect of rocking-withering in processing Wuyi rougui tea on the metabolites in leaves was analyzed using GC-TOF-MS non-targeted metabonomics. Method Freshly plucked buds and leaves from Wuyi rougui tea bushes were processed according to the conventional northern Fujian technique with or without (CK) the rocking-withering procedure. The non-targeted metabolic analysis was performed to obtain data for differentiating the metabolites in the leaves by multivariate and univariate statistical analyses. A KEGG pathway enrichment analysis was conducted to identify the associated metabolic pathways. Result Compared with the conventional brief withering under the sun, the different metabolites of tea leaves without rocking (CK) and after rocking-withering were 21 and 14; meanwhile tea leaves after rocking-withering had 9 significantly upregulated and 3 downregulated metabolites compared to leaves without rocking (CK) . The most significant differentiations were found in the metabolites such as shikimic acid, citric acid, L-malic acid, raffinose, and maltose. Conclusion Rocking-withering stimulated the tea leaves to produce certain intermediate products in the tricarboxylic acid cycle and caused a reduction on shikimic acid. It was the metabolisms of amino acids and carbohydrates that were largely affected. The significantly reduced shikimic acid and increased phenylalanine in LY7M metabolite might play a role in the synthesis of aromatic compounds that give the oolong tea its characteristic sensory quality.
- Wuyi rougui tea /
- rocking-withering /
- non-targeted metabolomics /
- primary metabolites

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图 1 各样品代谢物的PCA分析

Figure 1. PCA analysis on metabolites in samples

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图 2 SYM与CK7M代谢物的OPLS-DA分析

Figure 2. OPLS-DA analysis on SYM and CK7M metabolites

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图 3 SYM与LY7M代谢物的OPLS-DA分析

Figure 3. OPLS-DA analysis on SYM and LY7M metabolites

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图 4 CK7M与LY7M代谢物的OPLS-DA分析

Figure 4. OPLS-DA analysis on CK7M and LY7M metabolites

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图 5 样品间差异代谢物的KEGG通路富集分析

Figure 5. KEGG pathway enrichment analysis on differential metabolites in samples

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图 6 晒青叶SYM与摊青叶CK7M间差异代谢物

SYM-1～SYM-6指晒青叶；CK7M-1～CK7M-6指对照组。

Figure 6. Differential metabolites between sample SYM and CK7M

SYM-1 to SYM-6: withered leaves.CK7M-1 to CK7M-6: the control group(non-rocked leaves).

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图 7 晒青叶SYM与做青叶LY7M间差异代谢物

SYM-1～SYM-6指晒青叶；LY7M-1～LY7M-6指做青叶。

Figure 7. Differential metabolites between sample SYM and LY7M

SYM-1 to SYM-6: withered leaves; LY7M-1 to LY7M-6: tea leaves by turned over.

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图 8 摊青叶CK7M与做青叶LY7M间差异代谢物

CK7M-1～CK7M-6指对照组；LY7M-1～LY7M-6指做青叶。

Figure 8. Differential metabolites between sample CK7M and LY7M

CK7M-1 to CK7M-6: CK; LY7M-1 to LY7M-6: rocking-withered leaves.

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表 1 样品间差异代谢物的筛选

Table 1. Differential metabolites in samples

代谢产物 Metabolites	保留时间 Retention time/min	SYM & CK7M	SYM & LY7M	CK7M & LY7M	参考 Reference
丙氨酸 Alanine	8.02	↑	↑	—	标品 Standard sample
硫磺酸 Sulfuric acid	8.91	↑	—	↓	标品 Standard sample
异亮氨酸 Isoleucine	9.15	↑	—	—	标品 Standard sample
缬氨酸 Valine	9.61	↑	—	—	标品 Standard sample
琥珀酸 Succinic acid	10.99	↑	↑	↑	标品 Standard sample
L-苹果酸 L-Malic acid	13.26	↑	↑	—	标品Standard sample
3-己烯二酸 3-Hexenedioic acid	13.49	↑	↑	—	搜库 Database
苏糖酸 Threonic acid	14.09	↑	↑	↑	搜库 Database
肌酸 Creatine	15.15	↑	↑	—	搜库 Database
木糖1 Xylose 1	15.29	↑	↑	↑	搜库 Database
核糖 Ribose	15.46	↑	↑	↑	搜库 Database
苯丙氨酸 Phenylalanine	14.12	↑	↑	↑	标品 Standard sample
莽草酸 Shikimic acid	16.96	↑	—	↓	标品 Standard sample
柠檬酸 Ctric acid	17.05	↓	—	↑	标品 Standard sample
3-(4-羟基苯基)乳酸 DL-p-hydroxyphenyllactic acid	17.82	↓	—	—	搜库 Database
肌醇 Myo-inositol	19.75	↑	↑	—	搜库 Database
N-乙酰-β-D-甘露糖胺 N-acetyl-beta-D-mannosamine	19.89	↑	—	—	搜库 Database
D-鞘氨醇 D-erythro-sphingosine	22.65	↑	—	—	搜库 Database
二氢鞘氨醇 DL-dihydrosphingosine	23.21	↑	—	—	搜库 Database
熊果苷 Arbutin	23.79	↑	—	—	标品 Standard sample
棉子糖 Raffinose	30.03	↑	—	↓	搜库 Database
D-甘油酸 D-glyceric acid	11.17	—	↑	↑	搜库 Database
4-氨基丁酸1 4-aminobutyric acid 1	13.78	—	↑	↑	搜库Database
D-赤酮酸内酯 D-erythronolactone	14.58	—	↑	—	搜库 Database
橙皮素 Hesperitin	26.58	—	—	↑	标品 Standard sample
麦芽糖 Maltose	25.25	—	↑	—	标品 Standard sample
“↑”表示显著上升；“↓”表示显著下降；“—”表示无明显差异。 "↑" increase obviously, "↓"decrease obviously, "—"no obvious difference.

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