核桃炭疽病发生相关的酚类物质代谢分析

doi:10.11707/j.1001-7488.20211007

摘要/Abstract

摘要：

目的: 鉴定胶孢炭疽菌侵染核桃青皮后酚类物质种类以及变化规律，筛选潜在抗炭疽病有效成分，为探究核桃抵抗炭疽病发生机制提供参考。方法: 以‘香玲’和‘泰勒’2个品种核桃为试材，对其青皮体外接种胶孢炭疽菌，通过靶向代谢组学分析侵染后核桃青皮中酚类物质含量及相关变化。结果: 随着胶孢炭疽菌侵染天数增加，‘香玲’青皮病斑逐渐增大，在第6天发生明显变化，而‘泰勒’青皮病斑基本不变。‘香玲’和‘泰勒’青皮中酚类物质总量基本一致，划分为9大类，其中以原花青素/花青素、苯甲酸及其衍生物、黄酮醇和苯丙素类为主。‘香玲’中苯甲酸类最多，占60%以上，主要以没食子酸、丁香酸、鞣花酸和香草酸等为主；‘泰勒’中黄酮醇类最多，接近30%，主要以金丝桃苷、槲皮苷、扁蓄苷和杨梅苷等为主。炭疽菌侵染第6天，与‘香玲’相比，‘泰勒’青皮差异代谢物为60个，上调差异代谢物有52个，下调为8个，其中表儿茶素没食子酸酯、对香豆酸、原花青素B1/2/3、根皮苷、丁香酸、鞣花酸和阿魏酸等差异显著。分析炭疽菌侵染4~6天‘香玲’和‘泰勒’青皮差异代谢物发现，咖啡酸、柚皮素、（S）-圣草酚、扁蓄苷、槲皮苷和没食子酸6种物质出现明显变化，这可能与青皮病斑出现明显变化有关联。结论: 鉴定抗炭疽病品种‘泰勒’和感病品种‘香玲’130种酚类物质，代谢组学分析炭疽菌侵染后物质动态变化，筛选到原花青素B1/2/3、咖啡酸、柚皮素、（S）-圣草酚、扁蓄苷、槲皮苷和没食子酸等潜在抗炭疽病有效成分，可为后期开发炭疽病相关天然药物，探究炭疽病发生机制提供参考。

关键词: 代谢组学, 核桃, 炭疽病, 酚类物质

Abstract:

Objective: This study aimed to provide references for exploring the mechanism of walnut resistance to anthracnose. For this end, we identified 130 phenolic compounds after the infection of Colletotrichum gloeosprioides into the walnut husk, carried out absolute quantification of the compounds, analyzed their changes, and screened the potential effective components against anthracnose. Method: In this study, walnut (Juglans regia) cultivars, 'Xiangling' and 'Taile', were used as the materials, and the walnut husk were inoculated with C. gloeosporioides, then the contents and related changes of phenolic compounds in walnut husk after infection were analyzed by targeted metabonomics. Result: With the increase of days after infection of C. gloeosporioides, the plaques on 'Xiangling' husk gradually increased and changed significantly on the 6th day, while the plaques on 'Taile' husk remained basically unchanged. The total amount of phenolic compounds in 'Xiangling' and 'Taile' husk was basically the same. The compounds were divided into 9 categories, among which were mainly procyanidins/anthocyanins, benzoic acid derivatives, flavonols and phenylpropanoids. 'Xiangling' had the most benzoic acids, accounting for more than 60%, and they were mainly gallic acid, syringic acid, ellagic acid and vanillic acid. While 'Taile' contained the most flavonols, nearly 30%, and they were mainly hyperin, quercitrin, avicularin and myricetin. On the 6th day of anthracnose infection, comparing with 'Xiangling', there were 60 different metabolites in 'Taile', among which52 different metabolites were up-regulated and 8 different metabolites were down-regulated. Among them, epicatechin 3-O-gallate, 4-Hydroxycinnamic acid, proanthocyanidin B1/2/3, phlorizin, syringic acid, ellagic acid and ferulic acid had significant differences. In addition, 47 substances such askaempferol and m-coumaric acid were not detected. On analysis of the differential metabolites in the 'Xiangling' and 'Taile' husk infected for 4-6 days with anthracnose, it was found that 6 compounds of caffeic acid, naringenin, eriodictyol, avicularin, quercetin and gallic acid changedobviously, which might be related to the significant changes in husk plaques. Conclusion: We have identified 130 phenolic compounds in the anthracnose resistant cultivar 'Taile' and the susceptible cultivar 'Xiangling'. Potential effective components against anthracnose, including proanthocyanidin B1/2/3, caffeic acid, naringenin, eriodictyol, avicularin, quercetin and gallic acid, have been screened by analyzing the dynamic changes of substances after anthracnose infection by using metabolomics. This study would provide references for the later development of anthracnose-related natural medicines and for exploring the mechanism of anthracnose.

Key words: metabonomics, walnut, anthracnose, phenolic compounds

中图分类号:

S718.4

陈新,王敏,傅茂润,王贵芳,相昆,刘庆忠,焦文晓,张美勇,许海峰. 核桃炭疽病发生相关的酚类物质代谢分析[J]. 林业科学, 2021, 57(10): 71-80.

Xin Chen,Min Wang,Maorun Fu,Guifang Wang,Kun Xiang,Qingzhong Liu,Wenxiao Jiao,Meiyong Zhang,Haifeng Xu. Metabolic Analysis of Phenolic Compounds Associated with Walnut Anthracnose[J]. Scientia Silvae Sinicae, 2021, 57(10): 71-80.

图/表 6

图1

图2

图3

图4

表1

胶孢炭疽菌侵染6天后‘香玲’和‘泰勒’青皮间差异代谢物"

酚类物质 Phenolic compounds	‘泰勒’青皮(A) ‘Taile’ husk	‘香玲’青皮(S) ‘Xiangling’husk	A_S FC
表儿茶素没食子酸酯(-)-Epicatechin gallate	280.14±23.18 a	5.65±0.73 b	49.56
对香豆酸p-Coumaric acid	2 216.28±179.74 a	45.30±6.00 b	48.93
原花青素B2 Procyanidin B2	5 719.79±426.19 a	145.06±11.95 b	39.43
原花青素B1 Procyanidin B1	7 103.90±891.17 a	186.64±16.57 b	38.06
异樱花亭Isosakuranetin	13.40±0.32 a	0.36±0.03 b	36.77
樱花亭Sakuranetin	13.49±0.54 a	0.43±0.04 b	31.52
原花青素B3 Procyanidin B3	3 572.87±195.48 a	126.54±27.47 b	28.24
根皮苷Phlorizin	125.84±11.15 a	6.05±0.26 b	20.80
柚皮素Naringenin	102.10±7.86 a	5.60±0.47 b	18.24
表没食子儿茶素没食子酸酯Epigallocatechin gallate	37.70±3.49 a	2.62±0.48 b	14.40
咖啡酸Caffeic acid	13 428.80±527.38 a	963.40±76.81 b	13.94
表儿茶素Epicatechin	3 425.07±298.76 a	301.48±11.72 b	11.36
单咖啡酰酒石酸Caftaric acid	397.97±8.72 a	37.78±2.66 b	10.54
(S)-圣草酚Eriodictyol	78.14±3.08 a	8.30±0.90 b	9.41
芹菜素-7-O-β-D-吡喃葡萄糖苷(大波斯菊苷)Cosmosiin	2.84±0.13 a	0.30±0.05 b	9.39
杨梅素-3-O-半乳糖苷Myricetin 3-O-galactoside	5 147.34±305.84 a	558.18±53.50 b	9.22
扁蓄苷Avicularin	19 733.39±974.32 a	2 186.02±318.06 b	9.03
阿福豆苷Afzelin	1 059.41±51.54 a	117.27±7.27 b	9.03
儿茶素Catechin	15 254.58±995.10 a	1 756.35±63.42 b	8.69
7-羟基香豆素7-Hydroxycoumarin	226.63±16.21 a	30.13±2.84 b	7.52
香橙素Aromadendrin	556.69±44.16 a	78.33±8.14 b	7.11
花旗松素Taxifolin	1 277.19±95.28 a	182.89±15.37 b	6.98
杨梅苷Myricitrin	10 536.44±1 078.14 a	1 573.40±131.69 b	6.70
木犀草素Luteolin	51.98±5.52 a	7.88±0.48 b	6.60
异鼠李素-3-O-葡萄糖苷Isorhamnetin-3-O-glucoside	52.41±2.96 a	8.03±1.24 b	6.53
表没食子儿茶素(-)-Epigallocatechin	228.95±16.17 a	37.11±2.90 b	6.17
槲皮素Quercetin	198.21±11.88 a	36.23±3.61 b	5.47
没食子儿茶素(±)-Gallocatechin	532.04±30.67 a	103.08±6.87 b	5.16
紫云英苷Astragalin	555.28±22.14 a	109.19±9.22 b	5.09
桑色素Morin	189.66±11.76 a	42.24±2.34 b	4.49
瑞香素Daphnetin	7 859.44±136.79 a	1 878.56±50.63 b	4.18
山柰酚-3-O-芸香糖苷Kaempferol 3-O-rutinoside	39.51±1.38 a	9.56±1.45 b	4.13
金丝桃苷Hyperoside	67 206.66±3 097.02 a	17 156.62±836.22 b	3.92
隐绿原酸Cryptochlorogenic acid	108 225.60±7 123.72 a	29 877.01±3 384.46 b	3.62
氯化飞燕草素-3-O-葡萄糖苷Delphinidin-3-O-glucoside chloride	90 358.19±4 157.00 a	25 354.43±3 534.47 b	3.56
槲皮素3-O-葡萄糖酸苷Quercetin 3-O-glucuronide	1 091.22±92.93 a	319.10±36.04 b	3.42
紫苏醇Perillyl alcohol	1 096.52±69.28 a	348.78±43.45 b	3.14
槲皮苷Quercitrin	54 309.70±3 669.72 a	17 416.75±1 489.66 b	3.12
对羟基苯甲酸p-Hydroxybenzoic acid	22 831.83±697.61 a	7 800.56±244.09 b	2.93
氯化矢车菊素-3-O-芸香糖苷Cyanidin 3-O-rutinoside chloride	37.76±2.32 a	12.99±1.50 b	2.91
松柏醛Coniferaldehyde	1 071.67±44.49 a	436.55±30.59 b	2.45
水仙苷Narcissin	39.33±2.78 a	16.16±1.49 b	2.43
异鼠李素Isorhamnetin	1.98±0.13 a	0.83±0.11 b	2.38
二氢杨梅素Dihydromyricetin	65.07±4.89 a	28.03±2.93 b	2.32
牡荆素Vitexin	6.16±0.66 a	2.70±0.35 b	2.28
3, 4-二羟基苯甲醛3, 4-Dihydroxybenzaldehyde	1 089.56±40.36 a	484.21±8.20 b	2.25
香兰素Vanillin	2 113.21±85.33 a	1 061.65±80.67 b	2.00
樱桃甙Prunin	351.77±33.00 a	175.70±8.32 b	2.00
没食子酸甲酯Methyl gallate	554.25±37.72 b	1 222.48±122.18 a	0.45
丁香酸Syringic acid	84 040.53±1 839.92 b	188 943.60±4 031.86 a	0.44
乔松素(S)-Pinocembrin	2.86±0.37 b	7.24±0.87 a	0.39
阿魏酸Ferulic acid	4 728.12±298.64 b	13 909.08±430.68 a	0.34
2, 6-二羟基苯甲酸2, 6-Dihydroxybenzoic acid	17.36±0.61 b	63.72±4.42 a	0.27
鞣花酸Ellagic acid	15 159.33±910.95 b	75 323.18±4 279.05 a	0.20
杨梅素Myricetin	49.57±2.00	0
原花青素A2 Proanthocyanidin A2	27.67±255	0
橙皮苷Hesperidin	15.19±0.75	0
根皮素Phloretin	8.01±0.46	0
刺芒柄花苷Ononin	0.18±0	0
1, 3-二咖啡酰奎宁酸(洋蓟素)1, 3-Dicaffeoylquinic acid	0	1.97±0.06
毛蕊异黄酮Calycosin	0	20.32±1.39

表1

图5

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