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

doi:10.17521/cjpe.2016.0161

Abstract

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.

http://jtp.cnki.net/bilingual/detail/html/ZWSB201702007

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

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[J]. Chin J Plant Ecol, 2017, 41(2): 219-230.

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URL: https://www.plant-ecology.com/EN/10.17521/cjpe.2016.0161

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

Figures/Tables 7

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)

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)

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.

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.

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.

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

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^*

References 48

[1]	Adom KK, Liu RH (2002). Antioxidant activity of grains.Journal of Agricultural and Food Chemistry, 50, 6182-6187.
[2]	Agrawal AA (2001). Phenotypic plasticity in the interaction and evolution of species.Science, 294, 321-326.
[3]	Baque MA, Lee EJ, Paek KY (2010). Medium salt strength induced changes in growth, physiology and secondary metabolite contentin adventitious roots of Morinda citrifolia: The role of antioxidant enzymes and phenylalanine ammonia lyase.Plant Cell Reports, 29, 685-694.
[4]	Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochemistry, 72, 248-254.
[5]	Brandolini A, Castoldi P, Plizzari L, Hidalgo A (2013). Phenolic acids composition, total polyphenols content and antioxidant activity of Triticum monococcum, Triticum turgidum and Triticum aestivum: A two-years evaluation.Journal of Cereal Science, 58, 123-131.
[6]	Campos-Vargas R, Saltveit ME (2002). Involvement of putative chemical wound signals in the induction of phenolic metabolism in wounded lettuce.Physiologia Plantarum, 114, 73-84.
[7]	Chaman ME, Copaja SV, Argandoña VH (2003). Relationships between salicylic acid content, phenylalanine ammonialyase (PAL) activity, and resistance of barley to aphid infestation.Journal of Agricultural and Food Chemistry, 51, 2227-2231.
[8]	Dicke M, van Poecke RM, de Boer JG (2003). Inducible indirect defence of plants: From mechanisms to ecological functions.Basic and Applied Ecology, 4, 27-42.
[9]	Dixon RA, Paiva NL (1995). Stress induced phenylpropanoid metabolism.Plant Cell, 7, 1085-1097.
[10]	Fu JM, Huang BG (2001). Involvement of antioxidants and lipid peroxidation in the adaptation of two cool-season grasses to localized drought stress.Environmental and Experimental Botany, 45, 105-114.
[11]	Giannopolitis CN, Ries SK (1977). Superoxide dismutases I. Occurrence in higher plants.Plant Physiology, 59, 309-314.
[12]	Gill SS, Tuteja N (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants.Plant Physiology and Biochemistry, 48, 909-930.
[13]	Gogoi R, Singh DV, Srivastava KD (2001). Phenols as a biochemical basis of resistance in wheat against Karnal bunt.Plant Pathology, 50, 470-476.
[14]	Haruta M, Pedersen JA, Constabel CP (2001). Polyphenol oxidase and herbivore defense in trembling aspen (Populus tremuloides): cDNA cloning, expression, and potential substrates.Physiologia Plantarum, 112, 552-558.
[15]	Hu ZH, Yang D, Shen YB (2009). Difference of phenolic contents in leaves of Populus simonii × P. pyramidalis “Opera 8277” cuttings induced by various damages. Acta Botanica Boreali-Occidentalia Sinica, 29, 332-337. (in Chinese with English abstract)[胡增辉, 杨迪, 沈应柏 (2009). 不同损伤形式诱导合作杨叶片中酚类物质含量的差异. 西北植物学报, 29, 332-337.]
[16]	Jiang Y, Joyce DC (2003). ABA effects on ethylene production, PAL activity, anthocyanin and phenolic contents of strawberry fruit.Plant Growth Regulation, 39, 171-174.
[17]	Kessler A, Baldwin IT (2002). Plant responses to insect herbivory: The emerging molecular analysis.Annual Review of Plant Biology, 53, 299-328.
[18]	Kestring D, Menezes LCCR, Tomaz CA, Lima G, Rossi MN (2009). Relationship among phenolic contents, seed predation, and physical seed traits in Mimosa bimucronata plants.Journal of Plant Biology, 52, 569-576.
[19]	Koussevitzky S, Ne’eman E, Harel E (2004). Import of polyphenol oxidase by chloroplasts is enhanced by methyl jasmonate.Planta, 219, 412-419.
[20]	Kováčik J, Klejdus B, Hedbavny J, Bačkor M (2010). Effect of copper and salicylic acid on phenolic metabolites and free amino acids in Scenedesmus quadricauda (Chlorophyceae).Plant Science, 178, 307-311.
[21]	Lachman J, Amouz K, Šulc M, Orsák M, Pivec V, Hejtmánková A, Dvorák P, Cepl J (2009). Cultivar differences of total anthocyanins and anthocyanidins in red and purple- fleshed potatoes and their relation to antioxidant activity.Food Chemistry, 114, 836-843.
[22]	Latha P, Anand T, Ragupathi N, Prakasam V, Samiyappan R (2009). Antimicrobial activity of plant extracts and induction of systemic resistance in tomato plants by mixtures of PGPR strains and Zimmu leaf extract against Alternaria solani.Biological Control, 50, 85-93.
[23]	Li YQ, Sun YJ, Zhang TH, Zhao AF, Lian J (2011). Structural characteristics of Artemisia frigid community at different succession stages in Horqin Sandy Land.Chinese Journal of Applied Ecology, 22, 1725-1730. (in Chinese with English abstract)[李衍青, 孙英杰, 张铜会, 赵爱芬, 连杰 (2011). 科尔沁沙地不同演替阶段冷蒿群落的结构特征. 应用生态学报, 22, 1725-1730.]
[24]	Lin KM, Ye FM, Lin Y, Li QS (2010). Research advances of phenolic functional mechanisms in soils and plants.Chinese Journal of Eco-Agriculture, 18, 1130-1137. (in Chinese with English abstract)[林开敏, 叶发茂, 林艳, 李卿叁 (2010). 酚类物质对土壤和植物的作用机制研究进展. 中国生态农业学报, 18, 1130-1137.]
[25]	Liu NN, Tian QY, Zhang WH (2014). Comparison of adaptive strategies to phosphorus-deficient soil between dominant species Artemisia frigida and Stipa krylovii in typical steppe of Nei Mongol.Chinese Journal of Plant Ecology, 38, 905-915. (in Chinese with English abstract)[刘娜娜, 田秋英, 张文浩 (2014). 内蒙古典型草原优势种冷蒿和克氏针茅对土壤低磷环境适应策略的比较. 植物生态学报, 38, 905-915.]
[26]	Liu R, Wang ZY, Li TT, Wang F, An J (2014). The role of chitosan in polyphenols accumulation and induction of defense enzymes in Pinus koraiensis seedlings.Chinese Journal of Plant Ecology, 38, 749-756. (in Chinese with English abstract)[刘冉, 王振宇, 李婷婷, 王芳, 安静 (2014). 壳聚糖对红松幼苗多酚积累和抗氧化防御酶的诱导作用. 植物生态学报, 38, 749-756.]
[27]	Matkowski A (2006). Plant Phenolic Metabolites as Antioxidants and Mutagenesis Inhibitors. IOS Press, Amsterdam.
[28]	Meng JF, Fang YL, Qin MY, Zhuang XF, Zhang ZW (2012). Varietal differences among the phenolic profiles and antioxidant properties of four cultivars of spine grape (Vitis davidii Foex) in Chongyi County (China).Food Chemistry, 134, 2049-2056.
[29]	Nicholson RL, Hammerschmidt R (1992). Phenolic compounds and their role in disease resistance.Annual Review of Phytopathology, 30, 369-389.
[30]	Nordberg J, Arner ES (2001). Reactive oxygen species, antioxidants, and the mammalian thioredoxin system.Free Radical Biology and Medicine, 31, 1287-1312.
[31]	Paul S, Gooding CB (2001). Molecular cloning and characterization of banana fruit polyphenol oxidase.Planta, 213, 748-757.
[32]	Puthoff DP, Smigocki AC (2007). Insect feeding-induced differential expression of Beta vulgaris root genes and their regulation by defense-associated signals.Plant Cell Reports, 26, 71-84.
[33]	Robbins RJ (2003). Phenolic acids in foods: An overview of analytical methodology.Journal of Agricultural and Food Chemistry, 51, 2866-2887.
[34]	Sharma P, Jha AB, Dubey RS, Pessarakli M (2012). Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions.Journal of Botany, 2012, 1-26.
[35]	Tan BK, Harris ND (1995). Maillard reaction products inhibit apple polyphenoloxidase.Food Chemistry, 53, 267-273.
[36]	van Poecke RMP, Dicke M (2004). Indirect defence of plants against herbivores: Using Arabidopsis thaliana as a model plant.Plant Biology, 6, 387-401.
[37]	Wang J, Yang C, Han WQ, Liu ML (2003). Effects on water- soluble carbohydrate of Artemisia frigida under different defoliation intensities.Acta Ecologica Sinica, 23, 908-913. (in Chinese with English abstract)[王静, 杨持, 韩文权, 刘美玲 (2003). 刈割强度对冷蒿可溶性碳水化合物的影响. 生态学报, 23, 908-913.]
[38]	Wang Q, Yan SC, Jin H, Wang YJ (2012). Systemic induction of defensive enzymes in needles of Larix gmelinii seedlings in wounded conditions.Journal of Northeast Forestry University, 40(9), 77-80. (in Chinese with English abstract)[王琪, 严善春, 金虎, 王艳军 (2012). 叶损伤对兴安落叶松防御酶活性的系统诱导. 东北林业大学学报, 40(9), 77-80.]
[39]	Wang SP, Li YH, Wang YF, Chen ZZ (2001). Influence of different stocking rates on plant diversity of Artemisia frigida community in inner mongolia steppe.Acta Botanica Sinica, 43, 89-96. (in Chinese with English abstract)[汪诗平, 李永宏, 王艳芬, 陈佐忠 (2001). 不同放牧率对内蒙古冷蒿草原植物多样性的影响. 植物学报, 43, 89-96.]
[40]	Wang W, Liu ZL, Hao DY, Liang CZ (1996). Research on the restoring succession of the degenerated grassland in Inner Mongolia II. Analysis of the restoring processes.Acta Phytoecologica Sinica, 20, 460-471. (in Chinese with English abstract)[王炜, 刘钟龄, 郝敦元, 梁存柱 (1996). 内蒙古草原退化群落恢复演替的研究II. 恢复演替时间进程的分析. 植物生态学报, 20, 460-471.]
[41]	Winkel-Shirley B (1999). Evidence for enzyme complexes in the phenylpropanoid and flavonoid pathways.Physiologia Plantarum, 107, 142-149.
[42]	Xu GH, Guan RF, Ye XQ, Chen JC, Liu DH (2008). Composition and distribution of phenolic acids in satsuma mandarin (Citrus unshiu Marc.) during maturity.Food Science, 29(2), 137-141. (in Chinese with English abstract)[徐贵华, 关荣发, 叶兴乾, 陈健初, 刘东红 (2008). 不同成熟期蜜桔中酚酸的组成与分布. 食品科学, 29(2), 137-141.]
[43]	Yan SC, Yuan HE, Wang Q, Wang YJ (2010). Quantitative changes of phenolic acids in Larix gmelinii needles induced by leaf damage.Chinese Journal of Applied Ecology, 21, 1000-1006. (in Chinese with English abstract)[严善春, 袁红娥, 王琪, 王艳军 (2010). 叶损伤诱导兴安落叶松针叶中10种酚酸的变化. 应用生态学报, 21, 1000-1006.]
[44]	Zhang JE, Liu WG, Chen JQ, Shi YC, Cai YF (2005). Effects of different cutting intensities on above- and underground growth of Stylosanthes guianensis.Chinese Journal of Applied Ecology, 16, 1740-1744. (in Chinese with English abstract)[章家恩, 刘文高, 陈景青, 施耀才, 蔡燕飞 (2005). 不同刈割强度对牧草地上部和地下部生长性状的影响. 应用生态学报, 16, 1740-1744.]
[45]	Zhang RM, Wang YZ, Hou P, Wen GS, Gao Y (2010). Physiological responses to allelopathy of aquatic stem and leaf extract of Artemisia frigida in seedling of several pasture plants. Acta Ecologica Sinica, 30, 2197-2204. (in Chinese with English abstract)[张汝民, 王玉芝, 侯平, 温国胜, 高岩 (2010). 几种牧草幼苗对冷蒿茎叶水浸提液化感作用的生理响应. 生态学报, 30, 2197-2204.]
[46]	Zhang RM, Zhang WG, Zuo ZJ, Li R, Wu JH, Gao Y (2014). Inhibition effects of volatile organic compounds from Artemisia frigida Willd. on the pasture grass intake by lambs.Small Ruminant Research, 121, 248-254.
[47]	Zhao K, Baoyin T (2014). Effect of seasonal grazing use on productivity of grassland community.Chinese Journal of Grassland, 36(1), 109-115. (in Chinese with English abstract)[赵康, 宝音陶格涛 (2014). 季节性放牧利用对典型草原群落生产力的影响. 中国草地学报, 36(1), 109-115.]
[48]	Zuo ZJ, Zhang RM, Wang Y, Wen GS, Hou P, Gao Y (2010). The composition of volatile organic compounds (VOCs) emitted from damaged Artemisia frigida Willd. plants and their effects on root growth and development of pasture plants. Acta Ecologica Sinica, 30, 5131-5139. (in Chinese with English abstract)[左照江, 张汝民, 王勇, 温国胜, 侯平, 高岩 (2010). 损伤冷蒿挥发性有机化合物(VOCs)成分分析及其对牧草根系生长发育的影响. 生态学报, 30, 5131-5139.]