Changes in Total Phenol and Some Enzymatic and Non-Enzymatic Antioxidant Activities of Rose-scented Geranium (Pelargonium graveolens) in Response to Exogenous Ascorbic Acid and Iron Nutrition

Document Type: Original Article


Department of Horticultural Sciences, Faculty of Agriculture, Lorestan University, PO box 465, Korramabad, Iran


The strong antioxidant activity of Pelargonium graveolens is well established. The question addressed in this study was whether different concentrations of exogenous ascorbic acid (AsA) and iron (Fe) could influence the antioxidant activity and total phenol content (TPC) of geranium. Thus, three levels of Fe (0, 20 and 40 µM) and three levels of AsA (0, 1 and 2 mM) in the nutrient solution were combined factorially based on a completely randomized design with six replications, and chlorophyll content, TPC, and antioxidant activities of the leaves were measured. The results showed that oil content, ascorbate peroxidase (APX), and catalase (CAT) activities were increased in leaf samples under Fe starvation, regardless of the AsA concentration. The highest peroxidase (POD) activity was observed in samples treated with 20 µM Fe and 1 mM AsA. The highest total chlorophyll content was produced in plants treated with 40 µM Fe along with 1 mM AsA. TPC was increased with an increase in Fe concentration. Despite the positive effect of AsA on the pigment contents, plants treated with AsA showed lower TPC under all Fe concentrations. In total, lower Fe nutrition increased oil content and reactive oxygen species (ROS) scavenging activity of geranium. AsA application increased oil content while decreased total phenol and antioxidant activity in this plant.

Graphical Abstract

Changes in Total Phenol and Some Enzymatic and Non-Enzymatic Antioxidant Activities of Rose-scented Geranium (Pelargonium graveolens) in Response to Exogenous Ascorbic Acid and Iron Nutrition


  • Increasing chlorophyll content in P. graveolens under 40µM iron (Fe) concentration with 1mM ascorbic acid (AsA).
  • Reducing total phenol content and antioxidant activity and increasing essential oil content of P. graveolens by AsA application.
  • Enhancing essential oil content and reactive oxygen species scavenging activity in P. graveolens under low Fe concentration.
  • Increasing antioxidant enzymes activity and essential oil content of Pelargonium graveolens by Fe starvation with AsA treatment.


Blokhina, O., Virolainen, E. and Fagerstedt, K.V. 2003. Antioxidants, oxidative damage and oxygen deprivation stress: A review. Annals of  Botany, 91: 179–194.

Boukhris, M., Bouaziz, M., Feki, I., Jemai, H., El Feki, A. and Sayadi, S. 2012. Hypoglycemic and antioxidant effects of leaf essential oil of Pelargonium graveolens L’Hér. in alloxan induced diabetic rats. Lipids in Health and Disease, 11 (81): 1-10.

Boukhris, M., Hadrich, F., Chtourou, H., Dhouib, A., Bouaziz, M. and Sayadi, S. 2015. Chemical composition, biological activities and DNA damage protective effect of Pelargonium graveolens L’Hér. essential oils at different phenological stages. Industrial Crops and Products, 74: 600–606.

Boukhris, M., Simmonds, M.S., Sayadi, S. and Bouaziz, M. 2013. Chemical composition and biological activities of polar extracts and essential oil of rose‐scented geranium, Pelargonium graveolens. Phytotherapy Research, 27(8): 1206–1213.

Burits, M., Asres, K. and Bucar, F. 2001. The antioxidant activity of the essential oils of Artemisia afra, Artemisia abyssinica and Juniperus procera. Phytotherapy Research, 15 (2): 103–108.

Bybordi, A. 2012. Effect of ascorbic acid and silicium on photosynthesis, antioxidant enzyme activity, and fatty acid contents in canola exposure to salt stress. Journal of Integrative Agriculture, 11 (10): 1610–1620.

Ćavar, S. and Maksimović, M. 2012. Antioxidant activity of essential oil and aqueous extract of Pelargonium graveolens L’Her. Food Control, 23: 263–267.

Chance, B. and Maehly, A. 1955. Assay of catalases and peroxidases. Methods in Enzymology, 2: 764–775.

Curie, C. and Briat, J.F. 2003. Iron transport and signaling in plants. Annual Review of Plant Biology, 54: 183–206.

Eid, R., Taha, L. and Ibrahim, M. 2010. Physiological properties studies on essential oil of Jasminum grandiflorum L. as affected by some vitamins. Ozean Journal of Applied Sciences, 3(1): 87–96.

Ghahremani-majd, H., Dashti, F., Dastan, D., Mumivand, H., Hadian, J. and Esna-Ashari, M. 2012. Antioxidant and antimicrobial activities of Iranian mooseer (Allium hirtifolium Boiss.) populations. Horticulture Environment, and Biotechnology, 53(2): 116–122.

Gill, S.S. and Tuteja, N. 2010. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry, 48 (12): 909–930.

Guo, Z., Tan, H., Zhu, Z., Lu, S. and Zhou, B. 2005. Effect of intermediates on ascorbic acid and oxalate biosynthesis of rice and in relation to its stress resistance. Plant Physiology and Biochemistry, 43: 955–962.

Hindt, M.N. and Guerinot, M.L. 2012. Getting a sense for signals: Regulation of the plant iron deficiency response. Biochimica et Biophysica Acta (BBA) Molecular Cell Research, 1823 (9): 1521–1530.

Kabir, A.H., Rahman, M.M., Haider, S.A. and Paul, N.K. 2015. Mechanisms associated with differential tolerance to Fe deficiency in okra (Abelmoschus esculentus Moench.). Environmental and Experimental Botany, 112: 16–26.

Latifi, A., Jeanjean, R., Lemeille, S., Havaux, M. and Zhang, C.C. 2005. Iron starvation leads to oxidative stress in Anabaena sp. strain PCC 7120. Journal of Bacteriology, 187: 6596–6598.

Lichtenthaler, H.K. 1987. Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes. Methods Enzymology, 148: 350–382.

MacAdam, J.W., Nelson, C.J. and Sharp, R.E. 1992. Peroxidase activity in the leaf elongation zone of tall fescue I. Spatial distribution of ionically bound peroxidase activity in genotypes differing in length of the elongation zone. Plant Physiology, 99: 872–878.

Machold, O. and Stephan, U. 1969. The function of iron in porphyrin and chlorophyll biosynthesis. Phytochemistry, 8: 2189–2192.

Marsh, Jr.H., Evans, H. and Matrone, G. 1963. Investigations of the role of iron in chlorophyll metabolism. II. Effect of iron deficiency on chlorophyll synthesis. Plant Physiology, 38: 638–642.

Miller, N.J. and Rice-Evans, C.A. 1997. The relative contributions of ascorbic acid and phenolic antioxidants to the total antioxidant activity of orange and apple fruit juices and blackcurrant drink. Food Chemistry, 60: 331–337.

Misra, A. and Srivastava, N. 1990. Iron nutrition related to growth and physiology of Japanese mint (Mentha arvensis L.). Proceedings of the International Congress of Plant Physiology, New Delhi, India, 15-20 February, p. 1156–1160.

Mojaat, M., Pruvost, J., Foucault, A. and Legrand, J. 2008. Effect of organic carbon sources and Fe2+ ions on growth and β-carotene accumulation by Dunaliella salina. Biochemical Engineering Journal, 39: 177–184.

Nakano, Y. and Asada, K. 1981. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant and Cell Physiology, 22(5): 867–880.

Pandey, V. and Patra, D. 2015. Crop productivity, aroma profile and antioxidant activity in Pelargonium graveolens L’Hér. under integrated supply of various organic and chemical fertilizers. Industrial Crops and Products, 67: 257–263.

Pawar, N., Pai, S., Nimbalkar, M. and Dixit, G. 2011. RP-HPLC analysis of phenolic antioxidant compound 6-gingerol from different ginger cultivars. Food Chemistry, 126 (3): 1330–1336.

Qian, H., Peng, X., Han, X., Ren, J., Zhan, K. and Zhu, M. 2014. The stress factor, exogenous ascorbic acid, affects plant growth and the antioxidant system in Arabidopsis thaliana. Russian Journal of  Plant Physiology, 61: 467–475.

Ray, P.D., Huang, B.W. and Tsuji, Y. 2012. Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cellular Signalling, 24: 981–990.

Singleton, V.L., Orthofer, R. and Lamuela-Raventós, R.M. 1999. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu Reagent. Methods Enzymology, 299: 152–178.

Smirnoff, N. 1996. Botanical briefing: The function and metabolism of ascorbic acid in plants. Annals of Botany, 78 (6): 661–669.

Tewari, R.K., Hadacek, F., Sassmann, S. and Lang, I. 2013. Iron deprivation-induced reactive oxygen species generation leads to non-autolytic PCD in Brassica napus leaves. Environmental and Experimental Botany, 91: 74–83.

Vansuyt, G., Lopez, F., Inzé, D., Briat, J.F. and Fourcroy, P. 1997. Iron triggers a rapid induction of ascorbate peroxidase gene expression in Brassica napus. FEBS Letters, 410: 195–200.

Yeritsyan, N. and Economakis, C. 2002. Effect of nutrient solution's iron concentration on growth and essential oil content of oregano plants grown in solution culture. Acta Horticulturae, 576: 277–283.

Zancan, S., Suglia, I., La Rocca, N. and Ghisi, R. 2008. Effects of UV-B radiation on antioxidant parameters of iron-deficient barley plants. Environmental and Experimental Botany, 63: 71–79.