基于广靶代谢组学分析胡萝卜萜类代谢产物差异

doi:10.3864/j.issn.0578-1752.2024.16.012

Abstract

Abstract:

【Objective】 Terpenoid metabolites serve as primary indicators for assessing the quality of carrot flavor, which influence the taste and overall flavor quality of carrots significantly. This study compared the metabolites of flavor components in mature fleshy carrot roots exhibiting pronounced sweetness group and bitterness group, which could offer valuable insights for identifying key flavor components and enhancing the cultivation and exploitation of superior genetic resources.【Method】Efficient methods for detecting and identifying metabolites were established for carrot metabolomics research. Initially, a high-throughput metabolomic analysis platform was developed using UPLC-MS/MS to analyze terpene metabolites in carrots and to identify those influencing flavor quality. Differential metabolites were screened based on OPLS-DA results using VIP and log2FC values, with thresholds set at VIP>1 and fold changes of ≥2 and ≤0.5. 【Result】 A total of 50 terpenoid metabolites were screened out and exhibited significant differences, with 12 up-regulated and 2 down-regulated. Notable upregulated metabolites included Genipine (0.754), Japonicumin B (0.936), oxyphyllanene A (1.03), carvyl acetate (1.04), and 3-(3'-hydroxybutyl)-2, 4, 4-trimethylcyclohexan-2, 5-dienone (0.662). Dehydrodigitoxin (0.617) was significantly down-regulated. In terms of terpene content, those bitter carrots showed higher levels of Normelanothyrsin A (16.98), Curcumenol (17.57), and Genipin (15.32), while sweet carrots exhibited higher levels of Normelanothyrsin A (17.61), isothyrsin alcohol (16.01), and Curcuma alcohol (18.73). Top metabolites based on p-values included micrantholide (0.994), pyrodialdehyde (0.991), methyl (E)-5, 11-dihydroxy-4-(((Z)-2-2-enoyl methylbut-)foxy)-, 10-dimethylene-2-3 oxo-2 filling a, 4, 5, 8, 9, 10, 11, 11a-decahydrocyclodeca[b]furan-6-carboxylate (0.978), (3R)-3-hydroxy-ionone (0.974), and Blumenol A (0.969). Cluster analysis was employed to compare metabolite profiles between the two carrot groups, revealing significantly higher metabolite content in group 3 than that in group -3. Differential metabolite annotation and enrichment analysis were performed using the KEGG database.【Conclusion】Terpenoid metabolites, including Gennipine, Japonicumin B, oxyphyllanene A, carvyl acetate, Normelanothyrsin A, curcumenol, isocarbamol, dehydrodigitoxin, and micrantholide, were the primary compounds influencing the flavor of carrots. These compounds were believed to play a crucial role in determining the characteristic taste of carrots.

Key words: carrots（Daucus carota var. sativa Hoffm.）, broadly targeted metabolomics, flavor substance, terpene metabolites, differential metabolite, quality

QI XiaoYu, KONG XiaoPing, ZHOU HongWei, YAN XiangPing. Crucial Factors Impacting Carrot Flavor Analysis Based on Broad Target Metabolomics[J].Scientia Agricultura Sinica, 2024, 57(16): 3250-3263.

Figures/Tables 7

Table 1

Fig. 1

Fig. 2

Table 2

Terpenes detected by broad targeted metabolomics methods in carrot"

分类 Class	物质 Compound	B202	B207	H200	B176	B261	H41	VIP	Fold change	P-value
倍半萜 Sesquiterpenoids	薇甘菊内酯 Mikanolide	6.81E+05	1.83E+05	5.54E+05	1.44E+06	2.25E+06	7.02E+05	1.33E-02	1.00E+00	9.94E-01
	密叶辛木素 Confertifoline	9.74E+04	9.58E+04	9.52E+04	9.81E+04	9.21E+04	9.50E+04	3.49E-01	9.90E-01	7.67E-01
	甲黑素甲苷A Normelanothyrsin A	1.10E+07	1.14E+07	1.31E+07	1.03E+07	1.00E+07	1.36E+07	4.25E-01	1.05E+00	6.71E-01
	克里斯帕酮 Crispanone	1.08E+06	2.96E+06	1.76E+05	4.73E+05	2.64E+06	5.59E+04	1.06E+00	3.77E-01	1.60E-01
	香附可布酮 Kobusone	1.17E+05	1.10E+05	1.68E+05	1.70E+05	3.66E+05	1.28E+05	6.81E-02	9.85E-01	9.69E-01
	莪术醇 Curcumol	3.53E+06	4.69E+06	3.61E+06	5.35E+06	1.23E+07	8.82E+06	6.42E-01	1.08E+00	8.36E-01
	9α-羟基-11β,13-二氢扎鲁扎宁C 9alpha-hydroxy-11beta,13-dihydrozaluzanin C	7.73E+04	7.56E+04	7.52E+04	7.67E+04	7.09E+04	7.44E+04	6.41E-01	9.78E-01	5.93E-01
	氧叶烯A oxyphyllanene A	2.72E+06	2.62E+06	2.71E+06	2.53E+06	2.78E+06	2.85E+06	1.36E+00	1.03E+00	1.16E-01
	异呋喃醇二烯 Isofuranodiene (ISO2)	9.75E+05	1.42E+06	1.98E+06	1.11E+06	4.00E+06	2.16E+06	4.43E-02	1.09E+00	7.61E-01
	布卢门醇C Blumenol C	7.91E+05	2.26E+05	5.12E+05	1.24E+06	3.08E+06	7.62E+05	2.44E-01	9.62E-01	8.96E-01
	土木香内酯 Micheliolide	4.41E+05	5.66E+05	9.19E+05	3.14E+05	3.06E+06	7.02E+05	7.39E-01	7.25E-01	5.03E-01
	寥二醛 polygodial	1.79E+05	2.37E+05	1.73E+05	2.71E+05	6.80E+05	4.50E+05	7.15E-01	1.00E+00	9.91E-01
	1β-羟基-α-香附酮 1β-Hydroxy-α-cyperone (ISO6)	2.46E+06	2.40E+06	2.76E+06	2.21E+06	2.49E+06	2.54E+06	4.91E-01	1.02E+00	6.18E-01
	吉马酮 Germacrone	5.22E+05	1.34E+06	1.68E+06	7.42E+05	3.51E+05	8.27E+05	2.15E-01	9.76E-01	9.40E-01
	甲基(E)-5,11-二羟-4-((Z)-2-甲基丁-2-烯酰氧基）-3,10-二亚甲基-2-氧代 -2,3,3a,4,5,8,9,10,11,11a-十氢环癸并[b]呋喃-6-羧酸酯 Methyl (E)-5,11-dihydroxy-4-(((Z)-2-methylbut-2-enoyl)oxy)-3,10-dimethylene -2-oxo-2,3,3a,4,5,8,9,10,11,11a-decahydrocyclodeca[b]furan-6-carboxylate	2.23E+05	1.17E+05	1.53E+05	1.84E+05	5.87E+05	7.87E+05	4.23E-01	9.87E-01	9.78E-01
	异香附醇 Isocyperol	7.67E+05	2.71E+05	3.19E+05	1.01E+05	1.01E+07	3.92E+05	5.77E-01	9.37E-01	9.11E-01
	原莪术醇 Procurcumenol (ISO1)	2.09E+06	1.96E+06	3.00E+06	1.45E+06	3.41E+06	2.48E+06	1.05E+00	1.25E+00	3.61E-01
	3-(3'-羟基丁基)-2,4,4-三甲基环己烷-2,5-二烯酮 (R)-3-(3'-Hydroxybutyl)-2,4,4-trimethylcyclohexa-2,5-dienone	7.78E+04	6.19E+04	1.06E+05	1.48E+05	2.10E+05	2.60E+05	1.12E+00	6.62E-01	5.53E-01
	11-O-β-D-葡萄糖苷3-羟基-7,8-二氢-β-紫罗兰醇 11-O-β-D-Glucoside of 3-hydroxy-7,8-didehydro-β-ionol (ISO2)	5.24E+04	7.32E+04	8.83E+04	5.26E+04	1.74E+05	1.02E+05	1.38E-01	1.08E+00	7.19E-01
倍萜 Monoterpenoids	乙酸香芹酯 Carvylacetate	3.71E+05	4.38E+05	4.18E+05	5.01E+05	1.23E+06	7.78E+05	6.19E-01	1.04E+00	9.03E-01
	去氢催吐萝芙木醇 Dehydrovomifoliol	3.35E+05	4.61E+05	2.96E+05	9.11E+05	1.91E+05	9.62E+05	6.66E-01	6.30E-01	3.40E-01
	α-紫罗兰酮 α-Ionone	1.27E+05	1.73E+05	1.70E+05	1.56E+05	1.51E+05	1.60E+05	6.52E-01	1.03E+00	5.77E-01
	11-甲氧基维布尔替纳尔 11-methoxy viburtinal	4.99E+04	4.58E+04	9.71E+04	4.17E+04	6.47E+04	6.86E+04	3.63E-02	7.75E-01	6.22E-01
	大车前草苷 Majoroside	5.31E+05	9.27E+05	1.45E+06	4.45E+05	3.39E+05	6.86E+05	2.08E+00	1.96E+00	1.39E-02
	7-脱氧马钱苷酸 7-Deoxyloganic acid	1.02E+06	1.63E+06	4.40E+05	4.24E+05	1.59E+05	8.41E+05	1.39E+00	6.78E-01	5.80E-01
	脱氧斯坦硫苷 Deoxystansioside	2.91E+05	2.67E+05	5.82E+05	2.39E+05	3.59E+05	2.61E+05	4.61E-01	9.59E-01	8.90E-01
	3-羟基-紫罗兰酮 3-(6''-丙二酰)葡萄糖苷 3-hydroxy-beta-ionone 3-(6''-Malonyl) glucoside	2.91E+05	2.73E+05	5.68E+05	2.47E+05	3.65E+05	2.74E+05	5.70E-01	9.38E-01	8.28E-01
	丙二酰葡萄糖苷 Malonyl glucoside	2.93E+05	2.65E+05	5.51E+05	2.32E+05	3.38E+05	2.71E+05	4.00E-01	9.79E-01	9.47E-01
	京尼平 Genipin	2.49E+05	4.37E+06	4.71E+06	4.12E+06	4.87E+06	4.72E+06	8.05E-01	7.54E-01	4.69E-01
	紫苏子醇 Perillyl alcohol	6.57E+04	6.54E+04	1.14E+05	3.54E+04	3.49E+05	3.27E+05	4.55E-01	7.06E-01	6.73E-01
	3-氧-a-紫罗兰醇 3-(6''-丙二酰)葡萄糖苷 3-oxo-Alpha-ionol 3'-(6''-Malonyl) glucoside	2.64E+06	1.14E+06	1.59E+06	2.41E+06	3.08E+06	1.45E+06	1.10E+00	8.53E-01	2.69E-01
	布卢门醇 A Blumenol A	6.45E+04	8.59E+04	5.96E+04	8.76E+04	2.96E+05	1.22E+05	6.92E-01	1.01E+00	9.69E-01
	博夏林4-O-丙二酰葡糖苷 Boscialin 4-O-malonyl glucoside	6.72E+04	6.41E+04	2.44E+05	5.56E+04	3.51E+05	6.22E+05	4.34E-01	1.23E+00	6.19E-01
	(3R)-3-羟基-紫罗兰酮 3R-3-hydroxy-β-ionone	8.56E+04	1.17E+04	6.84E+04	2.72E+04	4.52E+05	2.28E+05	1.49E-01	1.02E+00	9.74E-01
	布卢门醇B丙二酰葡萄糖苷 Blumenol B malonyl Glucoside	1.22E+05	3.48E+04	2.70E+05	2.41E+04	3.30E+05	3.56E+05	1.09E+00	6.28E-01	3.11E-01
	二氢伏米叶醇4-O-丙二酰葡萄糖苷 Dihydrovomifoliol 4-O-malonyl glucoside	1.82E+05	3.48E+05	3.97E+05	2.70E+05	1.61E+06	1.51E+05	4.65E-01	1.87E+00	2.30E-01
	丙二酰拜占素苷A Malonyl byzantionoside A	3.65E+04	5.93E+04	9.53E+04	1.29E+04	1.27E+05	2.68E+05	3.50E-01	7.29E-01	4.91E-01
	可乔醇C Corchoionol C	1.37E+05	1.28E+05	1.69E+05	9.18E+04	1.26E+05	1.62E+05	4.59E-01	9.04E-01	6.70E-01
	丙二酰长寿花糖苷 Malonyl roseoside	6.93E+04	4.96E+04	1.29E+05	4.32E+04	1.86E+05	5.00E+04	1.16E+00	1.26E+00	4.78E-01
二萜 Ditepenoids	顺式泛醇 cis-Abienol	2.33E+04	3.00E+04	1.30E+05	2.51E+04	3.22E+04	1.56E+05	7.18E-01	1.05E+00	9.12E-01
	脱氢松香酸 Dehydroabietic acid	2.27E+06	3.48E+06	9.86E+05	9.76E+05	5.46E+05	2.69E+06	1.41E+00	6.73E-01	5.72E-01
	松香酸 Abietic acid	6.52E+04	1.23E+05	4.30E+04	5.86E+04	1.45E+05	3.37E+04	6.02E-01	2.57E+00	1.82E-01
	海松酸 Pimaric acid	6.09E+04	3.57E+04	1.00E+05	3.21E+04	1.28E+05	7.65E+04	1.56E-01	8.40E-01	7.53E-01
	异海松酸 Isopimaric acid	6.09E+04	3.57E+04	1.00E+05	3.21E+04	1.28E+05	7.65E+04	1.56E-01	8.40E-01	7.53E-01
	贝壳杉烯酸 Kaurenoic acid	2.18E+04	9.61E+03	6.07E+04	6.27E+03	6.00E+04	4.09E+04	1.20E+00	6.80E-01	2.68E-01
	维布萨宁J Vibsanin J	4.74E+04	5.50E+04	3.95E+04	6.17E+04	2.16E+05	2.70E+04	6.30E-01	1.57E+00	3.49E-01
三萜 Triterpene	齐墩果-12-烯-3,16-二酮 Olean-12-ene-3,16-dione	2.13E+04	5.40E+04	9.00E+00	1.71E+04	4.33E+04	1.70E+05	9.80E-01	5.87E-01	3.74E-01
三萜 Triterpene	粳米素B Japonicumin B	2.44E+04	3.78E+04	2.88E+04	2.46E+04	5.85E+04	2.56E+04	4.74E-01	9.36E-01	8.59E-01
萜类 Terpene	脱氢地芰普内酯 Dehydrololiolide	6.15E+04	7.41E+04	9.04E+04	5.12E+04	1.28E+05	1.05E+05	1.41E+00	6.17E-01	3.00E-01
萜类 Terpene	塞洛温A Sellowin A	3.42E+04	5.32E+04	1.15E+05	2.70E+04	9.76E+04	7.00E+04	1.52E+00	2.36E+00	4.01E-02

Table 2

Fig. 3

Fig. 4

Fig 5

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组 Group	引种号 Introduction number	根型 Root type	根表皮颜色 Root skin color	根肉色 Root-flesh color	根中心柱颜色 Root center column color	根长 Root length (cm)	根粗 Root thickness (cm)	单根重 Single weight (g)	口感 Taste
-3	B202	短圆柱 Short cylinder	黄色 Yellow	黄色 Yellow	黄色 Yellow	16.8	1.9	322.5	甜Sweet
	B207	短圆柱 Short cylinder	橘黄色 Aurantium	橘黄色 Aurantium	橘黄色 Aurantium	16.5	1.4	146.5	甜Sweet
	H200	短圆柱 Short cylinder	橘黄色 Aurantium	橘黄色 Aurantium	橘黄色 Aurantium	14.4	1.4	85.0	甜Sweet
3	B176	短圆柱 Short cylinder	黄色 Yellow	黄色 Yellow	黄色 Yellow	16.1	1.4	115.4	苦Bitter
	B261	短圆锥 Short cone	紫红色 Fuchsia	紫红色 Fuchsia	橘黄色 Aurantium	22.8	1.9	265.5	苦Bitter
	H41	短圆锥 Short cone	橘红色 Tangerine	红色 Red	黄色 Yellow	13.6	2.0	192.8	苦Bitter

Crucial Factors Impacting Carrot Flavor Analysis Based on Broad Target Metabolomics

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