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Effect of exogenous additives on oxidative stress and defense system of a tree: Zanthoxylum armatum DC. under in vitro conditions

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Toxicity related to oxidative stress has been a concern for mature, woody tree species with high phenolic content when placed under in vitro conditions, particularly at explant establishment stage. The study of biochemical factors responsible for oxidative stress (lipid peroxidation, reactive oxygen species like hydrogen peroxide) and the defense (antioxidant enzymes and non-enzymatic factors) that the plant offers, in the tree system under in vitro environment requires special consideration. This study focuses on the effect of exogenous additives [polyvinyl pyrrolidone (PVP), citric acid (CC), ascorbic acid (AA)] and initial dark treatment on oxidative stress, defense system, DNA oxidation, total chlorophyll content for the in vitro explant establishment of a medicinal tree Zanthoxylum armatum DC., for a duration of 1-month. Exogenous additives and darkness maintained a sustained level of lipid peroxidation hydrogen peroxide, oxidized DNA content and antioxidant enzymes (superoxide dismutase, catalase, and ascorbate peroxidase) in the modified treatment throughout the in vitro experimental period, in comparison to the control. Thus, establishing a relation between exogenous antioxidants and explant oxidative stress-free condition. This can further be utilized to prevent oxidative leaching of explant of mature and woody trees in vitro.

Key message

Explants of Z. armatum were able to survive under in vitro conditions for 1 month, with low levels of phenolic leaching and oxidative stress, when additives (2 g L−1 PVP, 100 mg L−1 CC, 100 mg L−1 AA) are added in the medium with initial dark treatment for 72 h.

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Fig. 1
Fig. 2



Polyvinyl pyrrolidone


Citric acid


Ascorbic acid


Photosynthetic photon flux density




Reactive oxygen species


Superoxide dismutase




Ascorbate peroxidase


Cetyl trimethylammonium bromide


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We are grateful to the Department of Biotechnology, Biotech Consortium India Limited, New Delhi, India, (DBT/JRF/14/AL/209 dated 1.07.2014) for providing Fellowship to Gyanmani Ekka. And, to Department of Science and Technology: Fund for Improvement of Science and Technology Infrastructure, India for providing fund (Sanction No. C. Dy. No. 2384/IFD/2014–15 dated 31.07.2014 and 06.08.2014) to the Department of School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India, where this work was performed.

Author information

Research idea, concept and designing: SKJ and AQ. Experimentation: GE. Manuscript drafting: GE. Critical evaluation: AQ. Approval of the manuscript: GE, SKJ and AQ.

Correspondence to Afaque Quraishi.

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Ekka, G., Jadhav, S.K. & Quraishi, A. Effect of exogenous additives on oxidative stress and defense system of a tree: Zanthoxylum armatum DC. under in vitro conditions. Plant Cell Tiss Organ Cult 140, 671–676 (2020). https://doi.org/10.1007/s11240-019-01759-4

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  • Antioxidant
  • Ascorbic acid
  • Citric acid
  • Phenolic-leaching
  • Poly-vinyl-pyrrolidone
  • Woody