老山芹层积前后种子差异代谢物分析

doi:10.13304/j.nykjdb.2023.0183

中国农业科技导报 ›› 2024, Vol. 26 ›› Issue (7): 37-49.DOI: 10.13304/j.nykjdb.2023.0183

老山芹层积前后种子差异代谢物分析

张永芳¹^,²(), 董世妍¹, 王佳轩¹, 郭绪虎¹, 张畅³, 王艳星¹, 王钰烨¹, 吴静凤¹, 鲍甜芳¹, 张宏发², 于萍², 李富恒²()

^1.山西大同大学, 山西大同 037009
^2.东北农业大学生命科学学院, 哈尔滨 150030
^3.东北石油大学, 黑龙江大庆 163000

收稿日期:2023-03-14 接受日期:2023-05-05 出版日期:2024-07-15 发布日期:2024-07-12
通讯作者: 李富恒
作者简介:张永芳 E-mail：zyf_208@163.com；
基金资助:
国家自然科学基金项目(31801872);山西省高等学校科技创新计划项目(2022L459);山西大同大学人文社科项目(2022YGZX005);大同市科技攻关项目(2020044);大同大学产学研项目(2020CXZ14)

Analysis of Differential Seed Metabolites Before and After Stratification of Heracleum moellendorffii Hance

Yongfang ZHANG¹^,²(), Shiyan DONG¹, Jiaxuan WANG¹, Xuhu GUO¹, Chang ZHANG³, Yanxing WANG¹, Yuye WANG¹, Jingfeng WU¹, Tianfang BAO¹, Hongfa ZHANG², Ping YU², Fuheng LI²()

^1.Shanxi Datong University，Shanxi Datong 037009，China
^2.College of Life Sciences，Northeast Agricultural University，Harbin 150030，China
^3.Northeast Petroleum University，Heilongjiang Daqing 163000，China

Received:2023-03-14 Accepted:2023-05-05 Online:2024-07-15 Published:2024-07-12
Contact: Fuheng LI

摘要/Abstract

摘要：

为深入探索老山芹种子休眠机理，以老山芹层积前后种子为研究对象，采用气相色谱-飞行时间质谱联用技术（gas chromatography-time of flight mass spectrometry，GC-TOF/MS）对其非层积种子（non-lamellarized seeds， NS）和层积后达到发芽状态种子（germination seeds， GS）的差异代谢物进行分析。结果表明，共检测到995种代谢产物，NS和GS的代谢物存在明显分离，差异代谢物有126种，其中极显著差异代谢物75种，包括35种上调，40种下调。对极显著差异代谢物所在代谢通路分析表明，涉及10条关键代谢通路。和弦图分析表明，差异代谢物中脂质类化合物（含类脂分子）与苯类化合物呈极显著正相关；苯丙烷类化合物（含聚酮类化合物）与有机氧类化合物呈极显著负相关。以上研究结果为探索老山芹种子成熟后萌发提供了代谢组学理论基础，为更好地研究老山芹种子后成熟及休眠机制提供了依据和支撑。

关键词: 老山芹, GC-TOF/MS, 代谢组学, 层积, 差异代谢物

Abstract:

In order to deeply explore the seed dormancy mechanism of Heracleum moellendorffii Hance， the different metabolites of non-lamellarized seeds （NS） and germination seeds after lamellarization （GS） were analyzed by GC-TOF/MS （gas chromatography-time of flight mass spectrometry）. The results showed that a total of 995 metabolites were detected， and there was a clear separation between NS and GS. There were 126 differential metabolites， among which 75 were extremely significant differential metabolites including 35 up-regulated and 40 down-regulated expressions. The metabolic pathways of the extremely significant differential metabolites were analysed and 10 key metabolic pathways were identified. The results of the chord diagram analysis showed that the lipids （including lipid-like molecules） was extremely significantly positive correlaed with benzenoids， while the benzenepropanoids （including polyketides） was extremely significantly negative correlation with organoxides. Above results provided theoretical basis of metabolomics for exploring the post-maturation germination of Heracleum moellendorffii Hance seeds， and researching the post-maturation and dormancy mechanisms of Heracleum moellendorffii Hance seeds.

Key words: Heracleum moellendorffii Hance, GC-TOF/MS, metabolomics, stratification, differential metabolites

中图分类号:

S567.23+9

张永芳, 董世妍, 王佳轩, 郭绪虎, 张畅, 王艳星, 王钰烨, 吴静凤, 鲍甜芳, 张宏发, 于萍, 李富恒. 老山芹层积前后种子差异代谢物分析[J]. 中国农业科技导报, 2024, 26(7): 37-49.

Yongfang ZHANG, Shiyan DONG, Jiaxuan WANG, Xuhu GUO, Chang ZHANG, Yanxing WANG, Yuye WANG, Jingfeng WU, Tianfang BAO, Hongfa ZHANG, Ping YU, Fuheng LI. Analysis of Differential Seed Metabolites Before and After Stratification of Heracleum moellendorffii Hance[J]. Journal of Agricultural Science and Technology, 2024, 26(7): 37-49.

图/表 11

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项目Item	参数Parameter
进样量Sample volume/μL	1
分流模式Front inlet mode	不分流模式Splitless mode
隔垫吹扫流速Front inlet septum purge flow/（mL·min^-1）	3
载气Carrier gas	氦气Helium
色谱柱Column	DB-5MS（30 m×250 μm×0.25 μm）
柱流速Column flow/（mL·min^-1）	1
柱箱升温程序Oven temperature ramp	50 ℃保持1 min；以10 ℃·min^-1升至310 ℃，保持8 min Hold on 1 min at 50 ℃， raise to 310 ℃ with a rate of 10 ℃·min^-1， hold on 8 min
前进样口温度Front injection temperature/℃	280
传输线温度Transfer line temperature/℃	280
离子源温度Ion source temperature/℃	250
质量范围Mass range （m/z）	50~500
电离电压Electron energy/eV	-70
扫描速率Acquisition rate	每秒20个光谱20 spectra per second
溶剂延迟Solvent delay/min	6.27

项目Item	参数Parameter
进样量Sample volume/μL	1
分流模式Front inlet mode	不分流模式Splitless mode
隔垫吹扫流速Front inlet septum purge flow/（mL·min^-1）	3
载气Carrier gas	氦气Helium
色谱柱Column	DB-5MS（30 m×250 μm×0.25 μm）
柱流速Column flow/（mL·min^-1）	1
柱箱升温程序Oven temperature ramp	50 ℃保持1 min；以10 ℃·min^-1升至310 ℃，保持8 min Hold on 1 min at 50 ℃， raise to 310 ℃ with a rate of 10 ℃·min^-1， hold on 8 min
前进样口温度Front injection temperature/℃	280
传输线温度Transfer line temperature/℃	280
离子源温度Ion source temperature/℃	250
质量范围Mass range （m/z）	50~500
电离电压Electron energy/eV	-70
扫描速率Acquisition rate	每秒20个光谱20 spectra per second
溶剂延迟Solvent delay/min	6.27

种类 Type	名称 Name	P值 P value	倍数变化 Fold change	Log₂ 倍数变化 Log₂ fold change
苯丙烷类化合物和聚酮类化合物类Phenylpropanoids and polyketides	新橙皮苷Neohesperidin	0.014 6	8.47×10⁰	3.08
	酪氨酸1 Tyrosine 1	0.005 1	2.10×10^-2	-5.57
	4-甲基伞形酮4-methylumbelliferone	0.026 2	4.32×10^-6	-17.80
苯类化合物类 Benzenoids	3，4-二羟基苯甲酸3，4-dihydroxybenzoic acid	0.006 6	5.18×10⁶	22.30
	间苯三酚Phloroglucinol	0.004 8	1.29×10⁶	20.30
	4-羟基苯甲酸4-hydroxybenzoic acid	0.026 6	6.69×10⁵	19.40
	对苯二甲酸Terephthalic acid	0.004 2	3.62×10⁵	18.50
	3-羟基苯甲酸3-hydroxybenzoic acid	0.030 0	1.89×10⁵	17.50
	苯乙醛2 Phenylacetaldehyde 2	0.015 9	2.57×10⁰	1.36
	3，4-二羟基扁桃酸3，4-dihydroxymandelic acid	0.000 0	2.48×10⁰	1.31
	对苯二酚Hydroquinone	0.008 9	6.42×10^-7	-20.60
核苷、核苷酸和类似物类Nucleosides， nucleotides， and analogues	尿苷1Uridine 1	0.010 7	1.47×10¹	3.87
	胞苷一磷酸脱胶产物Cytidine-monophosphate degr prod	0.000 6	7.13×10⁰	2.83
	单磷酸胞苷1 Cytidine-monophosphate 1	0.000 1	1.99×10^-5	-15.60
有机氮化合物类 Organonitrogen compounds	乙醇胺Ethanolamine	0.000 0	3.16×10^-1	-1.66
	1，3-二氨基丙烷1，3-diaminopropane	0.000 0	1.05×10^-1	-3.25
	3-氨基丙腈1 3-aminopropionitrile 1	0.000 9	3.36×10^-2	-4.90
有机酸及其衍生物类 Organic acids and derivatives	谷氨酸L-glutamic acid	0.003 9	4.12×10⁸	28.60
	环己基氨基磺酸1 Cyclohexylsulfamic acid 1	0.000 0	1.18×10⁶	20.20
	2-酮丁酸2 2-ketobutyric acid 2	0.002 5	2.58×10⁰	1.37
	5-氨基戊酸3 5-aminovaleric acid 3	0.000 0	2.48×10⁰	1.31
	3-氨基异丁酸1 3-aminoisobutyric acid 1	0.000 0	2.66×10^-1	-1.91
	4-氨基丁酸1 4-aminobutyric acid 1	0.000 3	2.31×10^-1	-2.12
	甘氨酸2 Glycine 2	0.000 2	1.52×10^-1	-2.72
	3-氰基苯胺3-cyanoalanine	0.001 2	2.61×10^-2	-5.26
	丝氨酸1 Serine 1	0.000 3	2.30×10^-2	-5.44
	尿素Urea	0.030 6	2.52×10^-5	-15.30
	α-酮异己酸1 α-ketoisocaproic acid 1	0.001 1	1.26×10^-5	-16.30
	邻磷酸丝氨酸1 O-phosphoserine 1	0.007 7	2.19×10^-6	-18.80
	邻乙酰丝氨酸1 O-acetylserine 1	0.019 7	2.13×10^-6	-18.80
	L-高丝氨酸1 L-homoserine 1	0.001 4	7.00×10^-7	-20.40
	邻苯三酚Pyrogallol	0.000 5	5.21×10^-7	-20.90
	L-半胱氨酸L-cysteine	0.004 3	2.36×10^-7	-22.00
	邻磷酸苏氨酸3 O-phosphonothreonine 3	0.026 1	2.07×10^-7	-22.20
	牛磺酸Taurine	0.001 1	1.96×10^-7	-22.30
	α-氨基己二酸α-aminoadipic acid	0.005 1	1.55×10^-7	-22.60
	柠檬酸Citric acid	0.006 3	4.88×10^-8	-24.30
	瓜氨酸1 Citrulline 1	0.006 7	1.03×10^-8	-26.50
有机氧化合物类 Organooxygen compounds	利比醇Ribitol	0.025 0	3.59×10⁸	28.40
	麦角糖Melezitose	0.048 0	9.70×10⁶	23.20
	莽草酸Shikimic acid	0.047 7	4.46×10⁶	22.10
	1，2-环己烷二酮4 1，2-cyclohexanedione 4	0.001 8	1.70×10⁶	20.70
	戊二醛2 Glutaraldehyde 2	0.027 4	1.18×10⁶	20.20
	丙酮3 Acetol 3	0.001 9	5.00×10¹	5.64
	半乳醇3 Galactinol 3	0.010 2	3.44×10⁰	1.78
	二羟丙酮Dihydroxyacetone	0.004 8	2.64×10⁰	1.40
	葡萄糖酸1 Gluconic acid 1	0.003 0	2.37×10^-1	-2.08
	D-甘油酸D-glyceric acid	0.000 0	2.11×10^-1	-2.24
	蔗糖酸Saccharic acid	0.006 8	4.25×10^-2	-4.55
	木糖醇Xylitol	0.013 8	7.61×10^-4	-10.40
	5’-甲基硫代腺苷1 5’-methylthioadenosine 1	0.036 8	6.65×10^-6	-17.20
	绿原酸1 Chlorogenic acid 1	0.012 6	1.29×10^-6	-19.60
	1，5-脱水葡萄糖醇1，5-anhydroglucitol	0.039 9	3.23×10^-7	-21.60
	葡萄糖1 Glucose 1	0.039 7	4.76×10^-8	-24.30
有机杂环化合物类Organoheterocyclic compounds	5，6-二氢尿嘧啶1 5，6-dihydrouracil 1	0.042 5	2.24×10⁶	21.10
	吡咯-2-羧酸Pyrrole-2-carboxylic acid	0.008 8	2.10×10⁶	21.00
	硫辛酰胺2 Thioctamide 2	0.002 6	9.76×10⁰	3.29
	生物素Biotin	0.008 7	4.41×10⁰	2.14
	2-羟基吡啶2-hydroxypyridine	0.000 0	2.44×10⁰	1.29
有机杂环化合物类Organoheterocyclic compounds	去甲烟碱Nornicotine	0.020 4	2.95×10^-2	-5.08
	乳清酸Orotic acid	0.001 9	2.64×10^-2	-5.24
	3-吲哚丙酮酸3-indolepyruvic acid	0.000 1	8.43×10^-3	-6.89
	抗坏血酸Ascorbate	0.003 3	8.46×10^-7	-20.20
脂质和类脂分子类 Lipids and lipid-like molecules	癸酸Capric acid	0.012 7	7.10×10⁶	22.80
	天竺葵酸Pelargonic acid	0.000 3	3.05×10⁶	21.50
	棕榈油酸Palmitoleic acid	0.022 0	2.61×10⁶	21.30
	月桂酸Lauric acid	0.000 3	1.10×10⁶	20.10
	麦芽糖醇Maltitol	0.021 1	1.58×10²	7.30
	氢化可的松Hydrocortisone	0.000 1	1.16×10¹	3.53
	十五酸Pentadecanoic acid	0.003 7	5.77×10⁰	2.53
	己二酸Adipic acid	0.000 0	2.48×10⁰	1.31
	苹果酸L-malic acid	0.001 3	3.16×10^-1	-1.66
	别苏氨酸1 L-allothreonine 1	0.000 2	3.68×10^-2	-4.76
	十二醇Dodecanol	0.000 3	1.19×10^-6	-19.70
	3-羟基-3-甲基戊二酸3-hydroxy-3-methylglutaric acid	0.031 0	6.10×10^-7	-20.60

种类 Type	名称 Name	P值 P value	倍数变化 Fold change	Log₂ 倍数变化 Log₂ fold change
苯丙烷类化合物和聚酮类化合物类Phenylpropanoids and polyketides	新橙皮苷Neohesperidin	0.014 6	8.47×10⁰	3.08
	酪氨酸1 Tyrosine 1	0.005 1	2.10×10^-2	-5.57
	4-甲基伞形酮4-methylumbelliferone	0.026 2	4.32×10^-6	-17.80
苯类化合物类 Benzenoids	3，4-二羟基苯甲酸3，4-dihydroxybenzoic acid	0.006 6	5.18×10⁶	22.30
	间苯三酚Phloroglucinol	0.004 8	1.29×10⁶	20.30
	4-羟基苯甲酸4-hydroxybenzoic acid	0.026 6	6.69×10⁵	19.40
	对苯二甲酸Terephthalic acid	0.004 2	3.62×10⁵	18.50
	3-羟基苯甲酸3-hydroxybenzoic acid	0.030 0	1.89×10⁵	17.50
	苯乙醛2 Phenylacetaldehyde 2	0.015 9	2.57×10⁰	1.36
	3，4-二羟基扁桃酸3，4-dihydroxymandelic acid	0.000 0	2.48×10⁰	1.31
	对苯二酚Hydroquinone	0.008 9	6.42×10^-7	-20.60
核苷、核苷酸和类似物类Nucleosides， nucleotides， and analogues	尿苷1Uridine 1	0.010 7	1.47×10¹	3.87
	胞苷一磷酸脱胶产物Cytidine-monophosphate degr prod	0.000 6	7.13×10⁰	2.83
	单磷酸胞苷1 Cytidine-monophosphate 1	0.000 1	1.99×10^-5	-15.60
有机氮化合物类 Organonitrogen compounds	乙醇胺Ethanolamine	0.000 0	3.16×10^-1	-1.66
	1，3-二氨基丙烷1，3-diaminopropane	0.000 0	1.05×10^-1	-3.25
	3-氨基丙腈1 3-aminopropionitrile 1	0.000 9	3.36×10^-2	-4.90
有机酸及其衍生物类 Organic acids and derivatives	谷氨酸L-glutamic acid	0.003 9	4.12×10⁸	28.60
	环己基氨基磺酸1 Cyclohexylsulfamic acid 1	0.000 0	1.18×10⁶	20.20
	2-酮丁酸2 2-ketobutyric acid 2	0.002 5	2.58×10⁰	1.37
	5-氨基戊酸3 5-aminovaleric acid 3	0.000 0	2.48×10⁰	1.31
	3-氨基异丁酸1 3-aminoisobutyric acid 1	0.000 0	2.66×10^-1	-1.91
	4-氨基丁酸1 4-aminobutyric acid 1	0.000 3	2.31×10^-1	-2.12
	甘氨酸2 Glycine 2	0.000 2	1.52×10^-1	-2.72
	3-氰基苯胺3-cyanoalanine	0.001 2	2.61×10^-2	-5.26
	丝氨酸1 Serine 1	0.000 3	2.30×10^-2	-5.44
	尿素Urea	0.030 6	2.52×10^-5	-15.30
	α-酮异己酸1 α-ketoisocaproic acid 1	0.001 1	1.26×10^-5	-16.30
	邻磷酸丝氨酸1 O-phosphoserine 1	0.007 7	2.19×10^-6	-18.80
	邻乙酰丝氨酸1 O-acetylserine 1	0.019 7	2.13×10^-6	-18.80
	L-高丝氨酸1 L-homoserine 1	0.001 4	7.00×10^-7	-20.40
	邻苯三酚Pyrogallol	0.000 5	5.21×10^-7	-20.90
	L-半胱氨酸L-cysteine	0.004 3	2.36×10^-7	-22.00
	邻磷酸苏氨酸3 O-phosphonothreonine 3	0.026 1	2.07×10^-7	-22.20
	牛磺酸Taurine	0.001 1	1.96×10^-7	-22.30
	α-氨基己二酸α-aminoadipic acid	0.005 1	1.55×10^-7	-22.60
	柠檬酸Citric acid	0.006 3	4.88×10^-8	-24.30
	瓜氨酸1 Citrulline 1	0.006 7	1.03×10^-8	-26.50
有机氧化合物类 Organooxygen compounds	利比醇Ribitol	0.025 0	3.59×10⁸	28.40
	麦角糖Melezitose	0.048 0	9.70×10⁶	23.20
	莽草酸Shikimic acid	0.047 7	4.46×10⁶	22.10
	1，2-环己烷二酮4 1，2-cyclohexanedione 4	0.001 8	1.70×10⁶	20.70
	戊二醛2 Glutaraldehyde 2	0.027 4	1.18×10⁶	20.20
	丙酮3 Acetol 3	0.001 9	5.00×10¹	5.64
	半乳醇3 Galactinol 3	0.010 2	3.44×10⁰	1.78
	二羟丙酮Dihydroxyacetone	0.004 8	2.64×10⁰	1.40
	葡萄糖酸1 Gluconic acid 1	0.003 0	2.37×10^-1	-2.08
	D-甘油酸D-glyceric acid	0.000 0	2.11×10^-1	-2.24
	蔗糖酸Saccharic acid	0.006 8	4.25×10^-2	-4.55
	木糖醇Xylitol	0.013 8	7.61×10^-4	-10.40
	5’-甲基硫代腺苷1 5’-methylthioadenosine 1	0.036 8	6.65×10^-6	-17.20
	绿原酸1 Chlorogenic acid 1	0.012 6	1.29×10^-6	-19.60
	1，5-脱水葡萄糖醇1，5-anhydroglucitol	0.039 9	3.23×10^-7	-21.60
	葡萄糖1 Glucose 1	0.039 7	4.76×10^-8	-24.30
有机杂环化合物类Organoheterocyclic compounds	5，6-二氢尿嘧啶1 5，6-dihydrouracil 1	0.042 5	2.24×10⁶	21.10
	吡咯-2-羧酸Pyrrole-2-carboxylic acid	0.008 8	2.10×10⁶	21.00
	硫辛酰胺2 Thioctamide 2	0.002 6	9.76×10⁰	3.29
	生物素Biotin	0.008 7	4.41×10⁰	2.14
	2-羟基吡啶2-hydroxypyridine	0.000 0	2.44×10⁰	1.29
有机杂环化合物类Organoheterocyclic compounds	去甲烟碱Nornicotine	0.020 4	2.95×10^-2	-5.08
	乳清酸Orotic acid	0.001 9	2.64×10^-2	-5.24
	3-吲哚丙酮酸3-indolepyruvic acid	0.000 1	8.43×10^-3	-6.89
	抗坏血酸Ascorbate	0.003 3	8.46×10^-7	-20.20
脂质和类脂分子类 Lipids and lipid-like molecules	癸酸Capric acid	0.012 7	7.10×10⁶	22.80
	天竺葵酸Pelargonic acid	0.000 3	3.05×10⁶	21.50
	棕榈油酸Palmitoleic acid	0.022 0	2.61×10⁶	21.30
	月桂酸Lauric acid	0.000 3	1.10×10⁶	20.10
	麦芽糖醇Maltitol	0.021 1	1.58×10²	7.30
	氢化可的松Hydrocortisone	0.000 1	1.16×10¹	3.53
	十五酸Pentadecanoic acid	0.003 7	5.77×10⁰	2.53
	己二酸Adipic acid	0.000 0	2.48×10⁰	1.31
	苹果酸L-malic acid	0.001 3	3.16×10^-1	-1.66
	别苏氨酸1 L-allothreonine 1	0.000 2	3.68×10^-2	-4.76
	十二醇Dodecanol	0.000 3	1.19×10^-6	-19.70
	3-羟基-3-甲基戊二酸3-hydroxy-3-methylglutaric acid	0.031 0	6.10×10^-7	-20.60

种类Type	上调Up-regulation	下调Down-regulation
有机酸及其衍生物类Organic acids and derivatives	4	17
脂质和类脂分子类Lipids and lipid-like molecules	8	4
有机杂环化合物类Organoheterocyclic compounds	5	4
苯类化合物类Benzenoids	7	1
核苷、核苷酸和类似物类Nucleosides， nucleotides， and alogues	2	1
有机氧化合物类Organooxygen compounds	8	8
有机氮化合物类Organonitrogen compounds	0	3
苯丙烷类化合物和聚酮类化合物类Phenylpropanoids and polyketides	1	2
其他类Others	21	29

老山芹层积前后种子差异代谢物分析

Analysis of Differential Seed Metabolites Before and After Stratification of Heracleum moellendorffii Hance

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