State of antioxidant systems and phenolic compounds’ production in Hypericum perforatum L. hairy roots

  • Oliver Tusevski
  • Jasmina Petreska Stanoeva
  • Marina Stefova
  • Mirko Spasenoski
  • Sonja Gadzovska SimicEmail author
Original Article


Hypericum perforatum hairy root clones (HR A–HR O) transformed with Agrobacterium rhizogenes A4 were evaluated for growth characteristics, phenolic compounds production, antioxidant/radical scavenging activity, antioxidant enzymes, and oxidative stress markers. The screening of growth characteristics revealed that HR clones were characterized with higher biomass accumulation, root elongation, and lateral branching in comparison to non-transformed roots. Significant increase of phenolics production in HR clones was related to phenylalanine ammonia lyase activity indicating an up-regulation of phenylpropanoid/flavonoid metabolism. Positive correlation of phenolics, flavonoids, and tannins with antioxidant assays indicated that these secondary metabolites significantly contributed to the antioxidant/radical scavenging properties of HR cultures. Regarding the enzymatic antioxidant state, an enhancement of superoxide dismutase activity in HR lines coincided with decrease in O 2 •− production rate, while ascorbate peroxidase and catalase greatly contributed to the reduction of H2O2 levels. The substantial accumulation of malondialdehyde in HR clones indicated an efficiency of antioxidant enzymes to reduce O 2 •− and H2O2 at levels that are not deleterious for membrane lipids. These results confirmed the involvement of an efficient antioxidant defense system in the response adjustment of H. perforatum HR cultures to transformation process. Two superior clones denoted as HR B and HR F were selected as fast-growing and high biomass yielding lines with up-regulated phenolic compounds’ production, antioxidant, and radical scavenging activity. Altogether, the best performing H. perforatum HR clones could be proposed as a prospective biotechnological system for scale-up production of antioxidant phenolic compounds.


Antioxidant activity Hairy roots Hypericum perforatum L. Phenolic compounds Reactive oxygen species 



Ascorbate peroxidase




Cupric ion-reducing antioxidant capacity


2,2-Diphenyl-1-picrylhydrazyl radical scavenging activity


Dry weight


Ferrous chelating activity


Fresh weight


Growth index


Hairy roots


Hydrogen peroxide scavenging activity


Lipid peroxidation inhibition




Nitric oxide scavenging activity


Number of secondary roots


Non-transformed roots

O2•− SA

Superoxide anion scavenging activity


Phenylalanine ammonia lyase


Polyphenol oxidase


Guaiacol peroxidase


Root length


Reactive oxygen species


Superoxide dismutase


Total condensed tannins


Total flavonoids


Total flavan-3-ols


Total phenolics


Total phenolic acids


Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.


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Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2019

Authors and Affiliations

  • Oliver Tusevski
    • 1
  • Jasmina Petreska Stanoeva
    • 2
  • Marina Stefova
    • 2
  • Mirko Spasenoski
    • 1
  • Sonja Gadzovska Simic
    • 1
    Email author
  1. 1.Institute of Biology, Faculty of Natural Sciences and MathematicsUniversity „Ss. Cyril and Methodius“SkopjeMacedonia
  2. 2.Institute of Chemistry, Faculty of Natural Sciences and MathematicsUniversity „Ss. Cyril and Methodius“SkopjeMacedonia

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