Chin J Plan Ecolo ›› 2017, Vol. 41 ›› Issue (2): 219-230.doi: 10.17521/cjpe.2016.0161

• Research Articles • Previous Articles     Next Articles

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 GAO1,*()   

  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:2017-03-16 Published:2017-02-10
  • Contact: Yan GAO
  • About author:

    KANG Jing-yao(1991-), E-mail:


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

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 <&lt;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"

Free phenolic acid
PPO 0.968** 0.861** -0.835** -0.871** 0.548 0.556 -0.194 0.723* 0.559 0.831**
PAL 0.859** 0.763** -0.785** -0.793** 0.641* 0.531 -0.069 0.690* 0.688* 0.752**
Bounded phenolic acid
PPO -0.635* 0.564 0.670* 0.742* -0.493 -0.050 0.579* 0.487 0.480 0.559
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"

Free phenolic acids
PPO 0.974** 0.953** 0.782** 0.861** 0.113 0.696* 0.797** 0.933** 0.711** 0.933**
PAL 0.678* 0.774** 0.478 0.511 0.254 0.346 0.483 0.618* 0.473 0.622*
Bounded phenolic acids
PPO -0.757** -0.581* -0.977** -0.912** 0.419 0.772** -0.938** -0.611* -0.529 -0.709**
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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