冷蒿酚酸及其抗氧化防御酶活性对机械损伤的响应

doi:10.17521/cjpe.2016.0161

植物生态学报 ›› 2017, Vol. 41 ›› Issue (2): 219-230.DOI: 10.17521/cjpe.2016.0161

冷蒿酚酸及其抗氧化防御酶活性对机械损伤的响应

刘盟盟¹, 贾丽¹, 程路芸¹, 张洪芹¹, 臧晓琳¹, 宝音陶格涛², 张汝民¹, 高岩^1,^*()

¹浙江农林大学亚热带森林培育国家重点实验室培育基地, 浙江临安 311300
²内蒙古大学生命科学学院, 呼和浩特 010021

收稿日期:2016-05-07 接受日期:2016-12-25 出版日期:2017-02-10 发布日期:2017-03-16
通讯作者: 高岩
作者简介:* 通信作者Author for correspondence (E-mail:sunzhiqiang1956@sina.com)
基金资助:
国家自然科学基金(31270756和31470704)、国家重点基础研究发展计划(973计划) (2014CB138805)和国家科技支撑计划项目(2011BAC07B01)

Responses of phenolic acid and defensive enzyme activities to mechanical damage in Artemisia frigida

Meng-Meng LIU¹, Li JIA¹, Lu-Yun CHENG¹, Hong-Qin ZHANG¹, Xiao-Lin ZANG¹, Taogetao BAOYIN², Ru-Min ZHANG¹, Yan GAO^1,^*()

¹The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang Agriculture and Forestry University, Lin’an, Zhejiang 311300, China

²College of Life Science, Inner Mongolia University, Hohhot 010021, China

Received:2016-05-07 Accepted:2016-12-25 Online:2017-02-10 Published:2017-03-16
Contact: Yan GAO
About author:KANG Jing-yao(1991-), E-mail: kangjingyao_nj@163.com。

摘要/Abstract

摘要：

为了探讨放牧过程中冷蒿(Artemisia frigida)酚酸及其抗氧化酶活性对牲畜采食和践踏损伤的响应, 该研究对盆栽冷蒿幼苗枝叶采用不同程度(轻度、中度和重度)机械损伤的方式模拟放牧强度, 利用高效液相色谱技术测定了机械损伤处理6 h后冷蒿枝叶和根系9种酚酸含量的变化; 同时对多酚氧化酶(PPO)、苯丙氨酸氨裂合酶(PAL)和抗氧化酶活性进行了测定, 分析了酚酸含量与PPO和PAL活性之间的相关性。结果表明: 轻度机械损伤可以诱导冷蒿枝叶和根系中PPO、PAL和抗氧化酶迅速做出响应, 活性显著升高; 随着机械损伤强度增加, 冷蒿枝叶游离态咖啡酸、丁香酸、阿魏酸和肉桂酸含量显著增加, 与对照相比中度损伤处理分别增加了150.4%、93.5%、154.4%和121.7%, 与PAL活性呈正相关关系; 游离态绿原酸和邻苯二酚含量减少, 与对照相比重度损伤处理后降低了91.1%和69.3%, 与PPO活性呈负相关关系; 与对照相比重度损伤处理下没食子酸和原儿茶酸分别增加了280.6%和215.7%。随损伤强度增加, 根系中9种游离态酚酸含量均显著增加, 其含量均小于叶片。机械损伤导致冷蒿枝叶中游离态和结合态酚酸总量增加, 根系中结合态酚酸减少。上述结果表明机械损伤首先诱导冷蒿体内应激防御系统中抗氧化酶和酚酸合成关键酶活性升高, 然后促进抗氧化物质酚酸的积累, 调节冷蒿植株内木质素合成以及醌类和单宁形成, 增强冷蒿抗机械损伤能力和提高冷蒿的耐牧性。

关键词: 冷蒿, 机械损伤, 酚酸, 抗氧化酶, 苯丙烷代谢

Abstract:

Aims The study aims at understanding the effects of feed intake and trample damage on the phenolic acid formation and antioxidant enzyme activities in Artemisia frigida, and elucidating the adaptive mechanisms in A. frigida to grazing in secondary metabolites and their related enzyme activities.
Methods We analyzed the phenolic acid content and the activities of polyphenol oxidase (PPO), phenylalanine ammonia-lyase (PAL) and protective enzymes in leaves and roots in A. frigida under three levels (light, moderate, and heavy) of manipulative grazing condition. The measurements of the 9 phenolic acid contents started after 6 h of the mechanical damage of the plants by using the high performance liquid chromatography (HPLC), and the enzyme activities in leaves and roots were measured by a spectrophotometry method.
Important findings The light damage treatment induced productions of PPO, PAL and significantly (p < 0.05) increased antioxidant enzyme activities in the leaves and roots of A. frigida. The contents of PPO, PAL and antioxidant enzymes increased with increasing intensity of mechanical damage. Compared to the control, the content of free caffeic, syringic, ferulic and cinnamic acid in the leaves A. frigida were significantly elevated (p < 0.05) by 150.4%, 93.5%, 154.4% and 121.7%, respectively. They were significantly (p < 0.05) positively correlated with PAL activity in the moderate damage treatment. The content of free chlorogenic acid and catechol decreased by 91.1%, and 69.3%, respectively, compared with the control they had a negative correlation with PPO activity in the heavy damage treatment. The contents of gallic and protocatechuic acids increased (p < 0.05) by 280.6% and 215.7%, respectively, in the heavy damage treatment. With increasing intensity of mechanical damage, the content of 9 free phenolic acids significantly increased in roots but the increasing range was less than the one in leaves. Mechanical damage induced an increasing trend in the total amount of free and bounded phenolic acids in the leaves but a decreasing trend in the total amount of bounded phenolic acids in the roots of A. frigida. The results indicated that mechanical damage could firstly induce an increase of antioxidant enzymes and key enzymes in phenolic metabolism in A. frigida, leading to the accumulation of antioxidant substances of phenolic acids, further regulate the biosynthesis of lignins, quinones and tannins, and then enhance the resistance to mechanical damage and improved the tolerance of A. frigida to grazing.

Key words: Artemisia frigida, mechanical damage, phenolic acid, antioxidant enzyme, phenylpropanoid pathway

刘盟盟, 贾丽, 程路芸, 张洪芹, 臧晓琳, 宝音陶格涛, 张汝民, 高岩. 冷蒿酚酸及其抗氧化防御酶活性对机械损伤的响应. 植物生态学报, 2017, 41(2): 219-230. DOI: 10.17521/cjpe.2016.0161

Meng-Meng LIU, Li JIA, Lu-Yun CHENG, Hong-Qin ZHANG, Xiao-Lin ZANG, Taogetao BAOYIN, Ru-Min ZHANG, Yan GAO. Responses of phenolic acid and defensive enzyme activities to mechanical damage in Artemisia frigida. Chinese Journal of Plant Ecology, 2017, 41(2): 219-230. DOI: 10.17521/cjpe.2016.0161

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链接本文: https://www.plant-ecology.com/CN/10.17521/cjpe.2016.0161

https://www.plant-ecology.com/CN/Y2017/V41/I2/219

图/表 7

表一机械损伤对冷蒿枝叶酚酸含量(μg·g-1fresh mass)的影响(平均值±标准误差,n=5)

Table 1 Effects of mechanical damage on the content of phenolic acids (μg·g-1fresh mass) in the leaves of Artemisia frigid (mean±SE, n=5)

表2 机械损伤对冷蒿根系酚酸含量(μg·g-1fresh mass)的影响(平均值±标准误差, n=5)

Table 2 Effects of mechanical damage on the content of phenolic acids (μg·g-1fresh mass) in the roots of Artemisia frigid (mean±SE, n=5)

图1 机械损伤对冷蒿苯丙氨酸氨裂合酶活性的影响(平均值±标准误差, n = 5)。不同大写字母表示冷蒿枝叶的差异显著, 不同小写字母表示根系的差异显著(p < 0.05, 根据LSD测验)。*, p < 0.05; **, p < 0.01。ns, 不显著。CK, 对照; LD为轻度损伤; MD为中度损伤; HD为重度损伤。

Fig. 1 Effect of mechanical damage on the activity of phenylalanine ammonia-lyase (PAL) in Artemisia frigid (mean ± SE, n = 5). Different uppercase letters indicate significant differences of leaves and different lowercase letters denote statistically significant differences of roots from Artemisia frigida (p < 0.05, according to LSD test). *, p < 0.05; **, p < 0.01; ns, non-significant. CK, control; LD, light damaging; MD, moderate damaging; HD, heavy damaging.

图2 机械损伤对冷蒿多酚氧化酶活性的影响(平均值±标准误差, n = 5)。不同大写字母表示冷蒿枝叶的差异显著, 不同小写字母表示根系的差异显著(p < 0.05, 根据LSD测验)。*, p < 0.05; **, p < 0.01。ns, 不显著。CK, 对照; LD为轻度损伤; MD为中度损伤; HD为重度损伤。

Fig. 2 Effect of mechanical damage on the activity of polyphenol oxidase (PPO) in Artemisia frigid (mean ± SE, n = 5). Different uppercase letters indicate significant differences of leaves and different lowercase letters denote statistically significant differences of roots from Artemisia frigida (p < 0.05, according to LSD test). *, p < 0.05; **, p <<0.01; ns, non-significant. CK, control; LD, light damaging; MD, moderate damaging; HD, heavy damaging.

图3 机械损伤对冷蒿抗氧化酶活性的影响(平均值±标准误差, n = 5)。不同大写字母表示冷蒿枝叶的差异显著, 不同小写字母表示根系的差异显著(p < 0.05, 根据LSD测验)。*, p < 0.05; **, p < 0.01。ns, 不显著。CK, 对照; LD为轻度损伤; MD为中度损伤; HD为重度损伤。

Fig. 3 Effect of mechanical damage on activity of antioxidant enzyme in Artemisia frigid (mean ± SE, n = 5). Different uppercase letters indicate significant differences of leaves and different lowercase letters denote statistically significant differences of roots from Artemisia frigida (p < 0.05, according to LSD test). *, p < 0.05; **, p < 0.01; ns, non-significant. CK, control; LD, light damaging; MD, moderate damaging; HD, heavy damaging.

表3 冷蒿枝叶酚酸含量与酶活性之间相关性分析

Table 3 Correlation analysis between the content of phenolic acids and related enzyme activities in the leaves of Artemisia frigid

	酶 Enzyme	没食子酸 Gallic	原儿茶酸 Protocatechuic	绿原酸 Chlorogenic	邻苯二酚 Pyrocatechol	咖啡酸 Caffeic	丁香酸 Syringic	对香豆酸 P-coumaric	阿魏酸 Ferulic	肉桂酸 Cinnamic	合计 Sum
游离酚酸 Free phenolic acid	PPO	0.968^**	0.861^**	-0.835^**	-0.871^**	0.548	0.556	-0.194	0.723^*	0.559	0.831^**
游离酚酸 Free phenolic acid	PAL	0.859^**	0.763^**	-0.785^**	-0.793^**	0.641^*	0.531	-0.069	0.690^*	0.688^*	0.752^**
结合酚酸 Bounded phenolic acid	PPO	-0.635^*	0.564	0.670^*	0.742^*	-0.493	-0.050	0.579^*	0.487	0.480	0.559
结合酚酸 Bounded phenolic acid	PAL	-0.646^*	0.576^*	0.413	0.542	-0.489	0.126	0.572	0.551	0.617^*	0.558

表4 冷蒿根系酚酸含量与酶活性之间相关性分析

Table 4 Correlation analysis between the content of phenolic acids and related enzyme activities in the roots of Artemisia frigid

	酶 Enzyme	没食子酸 Gallic	原儿茶酸 Protocatechuic	绿原酸 Chlorogenic	邻苯二酚 Pyrocatechol	咖啡酸 Caffeic	丁香酸 Syringic	对香豆酸 P-coumaric	阿魏酸 Ferulic	肉桂酸 Cinnamic	合计 Sum
游离酚酸 Free phenolic acids	PPO	0.974^**	0.953^**	0.782^**	0.861^**	0.113	0.696^*	0.797^**	0.933^**	0.711^**	0.933^**
游离酚酸 Free phenolic acids	PAL	0.678^*	0.774^**	0.478	0.511	0.254	0.346	0.483	0.618^*	0.473	0.622^*
结合酚酸 Bounded phenolic acids	PPO	-0.757^**	-0.581^*	-0.977^**	-0.912^**	0.419	0.772^**	-0.938^**	-0.611^*	-0.529	-0.709^**
结合酚酸 Bounded phenolic acids	PAL	-0.540	-0.465	-0.725^**	-0.561	0.106	0.425	-0.789^**	-0.635^*	-0.544	-0.689^*

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冷蒿酚酸及其抗氧化防御酶活性对机械损伤的响应

Responses of phenolic acid and defensive enzyme activities to mechanical damage in Artemisia frigida

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