基于广泛靶向代谢组学解析福建省2个产地山金柑果实次生代谢物差异

Analysis of differences in secondary metabolites of Fortunella hindsii Swingle fruits from two production areas in Fujian based on widely targeted metabolomics

  • 摘要: 目的 基于广泛靶向代谢组学解析福建省2个产地山金柑果实代谢组差异和药用成分特征,为山金柑区域化的种质资源保护、品质认证、功能产品开发及废弃物高值化利用提供理论依据。方法 以福建省泉州市永春县和南平市松溪县2个主栽区的山金柑果实为试验材料,利用超高效液相色谱串联三重四极杆质谱(UPLC-MS/MS)技术,对2个产地的山金柑种子和去籽果实进行广泛靶向代谢组学分析。通过主成分分析(PCA)、正交偏最小二乘判别分析(OPLS-DA)筛选差异代谢物,并结合KEGG代谢通路富集分析代谢物积累模式及功能。结果 广泛靶向代谢组学分析结果显示,从2个产地山金柑的去籽果实和种子中共鉴定到1594种次生代谢物,第一主成分(PC1)和第二主成分(PC2)对数据集的解释率分别为57.60%和17.86%;在2个山金柑产地的不同果实组织部位中分别检测到1119和1072种差异代谢物,其中组织特异性差异代谢物884种。2个产地去籽果实中黄酮类和萜类化合物的峰面积均显著高于种子(P<0.05,下同);而种子中木脂素和香豆素、醌类化合物的峰面积显著高于去籽果实。KEGG代谢通路富集分析结果显示,2个产地山金柑不同果实组织部位中组织特异性差异代谢物均富集在花青苷生物合成通路上。33种核心差异代谢物包括10种生物碱、7种黄酮类、12种木脂素和香豆素、2种酚酸以及2种其他类化合物,有6种核心差异代谢物在2个产地的去籽果实中均呈高表达,其中包括木犀草素-6-C-葡萄糖苷-7-O-鼠李糖苷、荭草素-2''-O-鼠李糖苷、木犀草素-8-C-葡萄糖苷(荭草素)等,筛选出24种具有地理标志差异的代谢物,其中永春县山金柑去籽果实和种子地理标志差异代谢物分别有8和11种,松溪县山金柑去籽果实和种子分别有3和2种。结论 山金柑果实次生代谢物在不同组织部位间的差异大于地理来源差异,去籽果实主要富集黄酮类和萜类化合物,种子主要富集生物碱、木脂素和香豆素等防御性代谢物;筛选获得24种永春县与松溪县地理标记差异代谢物。

     

    Abstract: Objective This study aimed to investigate the metabolome differences and medicinal component characteristics of Fortunella hindsii Swingle fruits from two production areas in Fujian Province based on widely targeted metabolomics, so as to provide a scientific basis for regional germplasm conservation,quality certification, functional pro-duct development, and high-value utilization of waste.Method Fortunella hindsii Swingle fruits from two main production areas in Fujian Province—Yongchun County in Quanzhou City, and Songxi County in Nanping City—were used as experimental materials. A widely targeted metabolomic analysis of seeds and seed-removed fruits from both production areas was conducted using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were employed to screen differential metabolites, and KEGG pathway enrichment analysis was performed to elucidate accumulation patterns and functions of the metabolites.Result The widely targeted metabolomics analysis identified a total of 1594 se-condary metabolites from seed-removed fruits and seeds of Fortunella hindsii Swingle from the two production areas; the first principal component (PC1) explained 57.60% for the dataset, and the second principal component (PC2) explained 17.86% for the dataset. 1119 and 1072 differentially metabolites were detected in different tissue parts of the fruits from the two production areas, among which 884 were tissue-specific differential metabolites. The peak areas of flavonoids and terpenoids in seed-removed fruits from the two production areas were significantly higher than those in seeds (P<0.05, the same below); the peak areas of lignans, coumarins, and quinones were significantly higher in seeds than those in seed-removed fruits. KEGG pathway enrichment analysis indicated that the tissue-specific differential metabolites in tissue parts of fruits from the two production areas were enriched in the anthocyanin biosynthesis pathway. In the 33 core differential metabolites were 10 alkaloids, 7 flavonoids, 12 lignans and coumarins, 2 phenolic acids, and 2 other compounds. Among these, 6 core differential metabolites were consistently highly expressed in the seed-removed fruits from the two production areas, including luteolin-6-C-glucoside-7-O-rhamnoside, orientin-2''-O-rhamnoside, luteolin-8-C-glucoside (orientin). Thereby, 24 geographical-specific differential metabolites were identified: samples of seed-removed fruits and seeds from Yongchun County had 8 and 11 geographical-specific differential metabolites, respectively; samples of seed-removed fruits and seeds from Songxi County had 3 and 2 geographical-specific differential metabolites, respectively.Conclusion Differences in secondary metabolites of Fortunella hindsii Swingle fruits are greater between tissue parts than between geographical origins. Seed-removed fruits are primarily enriched with flavonoids and terpenoids, while seeds are mainly enriched with defensive metabolites such as alkaloids, lignans, and coumarins. A total of 24 geographical-specific differential metabolites between Yongchun County and Songxi County are identified through identification.

     

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