Biochemical changes in barberries during adventitious root formation: the role of indole-3-butyric acid and hydrogen peroxide

Ali Tehranifar, Saeed Mahmoody Tabar, Yahya Selahvarzi, Ahmad Balandary, Mahdiyeh Kharrazi


Peroxidase, polyphenol oxidase (PPO), phenolic compounds and total sugars (TS) were investigated during root formation in cuttings of Berberis vulgaris var. asperma (BVA) and Berberis thunbergii var. atropurpurea (BTA) treated with indole-3-butyric acid (IBA) and IBA+H2O2. Rooting was observed on BTA cuttings but not on BVA cuttings. The BTA cuttings treated with IBA and IBA+H2O2 showed higher rooting percentages, number of roots, and root length over the control. Those treated with IBA+H2O2 recorded the lowest peroxidase activity after planting. BTA cuttings treated with IBA+H2O2 showed the highest peroxidase activity at 50 d after planting; BVA cuttings under different treatments showed no significant difference for peroxidase activity at planting time or up to 80 d after planting. PPO activity for the BTA cuttings in the control treatment was lower than for other treatments during root formation. The cuttings in the IBA and IBA+H2O2 treatments showed increased PPO activity from 0 to 50 d after planting and a slight decrease in PPO activity from 60 to 80 d after planting. PPO activity for the BVA cuttings was significantly lower than for BTA during root formation. The BTA cuttings treated with IBA and IBA+H2O2 showed the highest phenolic compound content during root formation. The BVA cuttings displayed higher TS than BTA during the initial stage of root formation. A comparison of the anatomical structure of easy-to-root and difficult-to-root cuttings indicated that physical inhibitors did not affect the rooting capacity of BVA.


Berberis vulgaris var. asperma; Berberis thunbergii var. atropurpurea; peroxidase; polyphenol oxidase; phenolic compounds

Full Text:



Balandary A, Kafi M, 2001. Berberis production and processing. Zaban va Adab Press, Mashhad, Iran. [In Persian].

Beffa R, Martin HV, Pilet PE, 1990. In vitro oxidation of indoleacetic acid by soluble auxin-oxidase and peroxidases from maize roots. Plan Physiol 94: 485-491.

Bradford MM, 1976. A rapid and sensitive method for the quantification of microgram quanties of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254.

Ford YY, Bonham EC, Cameron RWF, Blake PS, Judd HL, Harrison-Murray RS, 2001. Adventitious rooting: examining the role of auxin in an easy- and a difficult-to-root plant. Plant Grow Regul 36: 149-159.

Gaspar T, Kevers C, Hausman JF, Berthon JY, Ripetti V, 1992. Practical uses of peroxidase activity as a predictive marker of rooting performance of micropropagated shoots. Agronomie 12: 757-765.

Gunes T, 2000. Peroxidase and IAA-oxidase activities during rooting in cuttings of three poplar species. Turk J Bot 24: 97-101.

Hartmann HT, Kester DE, Davies FT, 1989. Plant propagation: principles and practices. Prentice Hall.

Huystee RB, Cairns WL, 1982. Progreno and prospects in the use of peroxidase to cell development. Phytochem 21: 1843-1847.

Irigoyen JJ, Emerich DW, Sanchez-Dias M, 1992. Water stress induced changes in concentrations of proline and total soluble sugars in nodulated alfalfa (Medicago sativa) plants. Plant Physiol 15: 55-60.

Jayaprakasha GK, Singh RP, Sakariah KK, 2001. Antioxidant activity of grape seed (Vitis vinefera) extracts on peroxidation models in vitro. Food Chem 73: 285-290.

Kar M, Mishra D, 1976. Catalase, peroxidase and polyphenol oxidase activities during rice leaf senescence. Plant Physiol 57: 315-319.

Kaur S, Cheema SS, Chabra BR, Talwar KK, 2002. Chemical induction of physiological changes during adventitious root formation and bud break in grapevine cuttings. Plant Grow Regul 37: 63-68.

Kevresan S, Kovacevic B, Cirin-Novta V, Kuhajda K, Kandrac J, Pavlovic K, Grbovic L, 2007. Biochemical changes in cuttings of Robinia pseudoacacia after treatment with naphthenate. J Serb Chem Soc 72: 953-959.

Kibber H, Johnston ME, William RR, 2004. Adventitious root formation in cuttings of Backhousia citriodora F. Muell 1. Plant genotype, juvenility and characteristics of cuttings. Sci Hort 102: 133-143.

Li SW, Xue L, Xu S, Feng H, An L, 2009a. Hydrogen peroxide acts as a signal molecule in the adventitious root formation of mung bean seedlings. Environ Exp Bot 65: 63-71.

Li SW, Xue L, Xu S, Feng H, An L, 2009b. IBA-induced changes in antioxidant enzymes during adventitious rooting in mung bean seedlings: the role of H2O2. Environ Exp Bot 66: 442-450.

Lubell JD, Brand MH, Lehrer JM, 2008. AFLP identification of Berberis thunbergii cultivars, inter-specific hybrids, and their parental species. J Hort Sci Biotech 83: 55-63.

Ludwig-Muller J, 2003. Peroxidase isoenzymes as markers for the rooting ability of easy-to root and difficult-to-root Gravillea species and cultivars of Protea obstusifolia (Proteaceae). In Vitro Cell Dev Biol Plant 39: 377-383.

Ozyuigit II, 2008. Phenolic changes during in vitro organogenesis of cotton (Gossypium hirsutum L.). Afr J Biotech 7: 1145-1150.

Rout GR, 2006. Effect of auxins on adventitious root development from single node cuttings of Camellia sinensis L. Kuntze and associated biochemical changes. Plant Grow Regul 48: 111-117.

Satish J, Raveendran P, Rokade ND, 2008. Changes in polyphenol oxidase activity during rooting of hardwood cuttings in three grape rootstocks under Indian condition. S Afr J Enol Vitic 29: 94-97.

Sebastiani L, Tognetti R, 2004. Growing season and hydrogen peroxide effects on root induction and development in Olea europaea L. (cvs 'Frantoio and Gentile di Larino') cuttings. Sci Hort 100: 75-82.

Sebastiani L, Tognetti R, Di paolo P, Vitagliano C, 2002. Hydrogen peroxide and indole-3-butyric acid effects on root induction and development in cuttings of Olea europaea L. (cv. Frantoio and Gentile di Larino). Adv Hort Sci 16: 7-12.

Sivaci A, Yalcim I, 2007. Determination of physiological changes in related to effect of exogenous indole-butyric acid and callus formation in some kinds of apple (Malus sylvestris M.) stem cuttings. Fen Bilimleri Dergisi 27: 1-12.

Strzeleka K, 2007. Anatomical structure and adventitious root formation in Rhododendron ponticum L. cuttings. Acta Sci Pol Hortorum Culture 6: 15-22.

Tehranifar A, 2003. Barberry growing in Iran. Acta Hort 620: 193-195.

Tomas P, Ravindra MB, 1999. Shoot tip culture in mango influence of medium, genotype, explants factors, season and decontamination treatments on phenolic exudation, explants survival and axinic culture establishment. J Hort Sci 72: 713-722.

Tworkoski T, Takeda F, 2007. Rooting response of shoot cuttings from three peach growth habits. Sci Hort 115: 98-100.

White R, 1994. Environmentally persistent alkylphenolic compounds are strogenic. Endochrinol 135: 175-182.

Yilaz H, Taskin T, Otludil B, 2003. Polyphenol oxidase activity during rooting in cuttings of grape (Vitis vinifera L.) varieties. Turk J Bot 27: 495-498.

DOI: 10.5424/sjar/2014122-3889