Document Type : Research Paper


1 Associate Professor, Department of Horticultural Sciences, Faculty of Agriculture, University of Bu-Ali Sina, Hamedan, Iran.

2 Former M.Sc. Student, Department of Horticultural Sciences, Faculty of Agriculture, Bu-Ali Sina University, Hamedan, Iran.

3 Assistant Professor, Department of Horticultural Sciences, Faculty of Agriculture, University of Bu-Ali Sina, Hamedan, Iran.


The phonological stage in which the abotic stress occurs is effective in determining the content of secondary metabolites in medicinal plants. This research deals with the effects of imposing drought stress at different phenological stages on growth and some physiological and phytochemical responses of Nepeta crispa L as a split plot design in Bu-Ali Sina University during 2019. Drought stress at three levels, including mild, moderate, and severe stress (with 80%, 60%, and 40% of field capacity, respectively) has been applied at two phenological stages (seedling establishment stage with 6 true leaves and pre-flowering stages). Results from statistical analysis show that drought stress has had a significant effect on all evaluated growth traits and phytochemical parameters of the plant including proline content, relative water content (RWC) of leaves, total phenols, flavonoids, tannins, antioxidant activities, and phenolic acids. Imposing drought stress at both phenological stages reduce plant growth, decreasing chlorophyll content and RWC. However, production of phenolic compounds and phenolic acids (including rosmarinic, chlorogenic and caffeic acids) at high level and accumulation of proline through imposing drought stress reduce the adverse effects of stress and increase the medicinal properties of the Nepeta crispa. Also, the effect of drought stress imposing at pre-flowering stage has been greater than its application in the seedling establishment stage. According to the results, it is recommended to impose moderate drought stress at pre-flowering stage to increase biosynthesis of some phytochemical compounds in Nepeta crispa medicinal plants.


Ahmadvand, H., Amiri, H., & Dalvand, H. (2015). Antioxidant properties of hydro-alcoholic extract and extract of nepeta crispa in Lorestan province. Hormozgan Medical Journal, 19(3), 172-179. (In Persian)
Ainsworth, E.A., & Gillespie, K.M. (2007). Estimation of total phenolic content and other oxidation substrates in plant tissues using Folin–Ciocalteu reagent. Nature protocols, 2(4),875-877.
Alhaithloul, H. A., Soliman, M. H., Ameta, K. L., El-Esawi, M. A., & Elkelish, A. (2020). Changes in Ecophysiology, Osmolytes, and Secondary Metabolites of the Medicinal Plants of Mentha piperita and Catharanthus roseus Subjected to Drought and Heat Stress. Biomolecules, 10(1), 43.
Alizadeh Ahmadabadi, A., & Khorasaninejad, S. (2017). The Effect of Humic acid Pretreatment on Germination of purple cornflower (Echinacea purpurea) plant under Drought and Salinity Conditions. Arid Biome Scientific and Research Journal, 6(2), 97-107. (In Persian)
Bahreininejad, B., Razmjou, J., & Mirza, M. (2013). Influence of water stress on morpho-physiological and phytochemical traits in Thymus daenensis. Internationa Journal of Plant production, 7(1), 151-166.
Bates, L. S., Waldren, R.., & Teare, I. (1973). Rapid determination of free proline for water-stress studies. Plant and Soil, 39(1), 205-207.
Farnad, N., Heidari, R., & Aslanipour, B. (2014). Phenolic composition and comparison of antioxidant activity of alcoholic extracts of Peppermint. Journal of Food Measurement and Characterization, 8(2), 113-121.
Figueroa-Pérez, M. G., Rocha-Guzmán, N. E., Pérez-Ramírez, I. F., Mercado-Silva, E., & Reynoso-Camacho, R. (2014). Metabolite profile, antioxidant capacity, and inhibition of digestive enzymes in infusions of peppermint (Mentha piperita) grown under drought stress. Journal of agricultural and food chemistry, 62(49), 12027-12033.            
Ghannadi, A., Aghazari, F., Mehrabani, M., Mohagheghzadeh, A., & Mehregan, I. (2010). Quantity and composition of the SDE prepared essential oil of Nepeta macrosiphon Boiss. Iranian Journal of Pharmaceutical Research, (2), 103-105.  (In Persian)  
Gupta, A., Rico-Medina, A., & Caño-Delgado, A. I. (2020). The physiology of plant responses to drought. Science, 368(6488), 266-269.
Johnston, K. L., Clifford, M. N., & Morgan, L. M. (2003). Coffee acutely modifies gastrointestinal hormone secretion and glucose tolerance in humans: glycemic effects of chlorogenic acid and caffeine. The American Journal of Clinical Nutrition, 78(4), 728-733.
Khorasaninejad, S., Soltanloo, H., Ramezanpour, S. S., Hadian, J., & Atashi, S. (2015). The effect of drought stress on the growth, essential oil yield and chemical composition of Lavender. Journal of Crops Improvement, 17(4), 979-988.       
Kulkarni, A. P., & Aradhya, S. M. (2005). Chemical changes and antioxidant activity in pomegranate arils during fruit development. Food chemistry, 93(2), 319-324.
Li, Y., Guo, C., Yang, J., Wei, J., Xu, J., & Cheng, S. (2006). Evaluation of antioxidant properties of pomegranate peel extract in comparison with pomegranate pulp extract. Food chemistry, 96(2), 254-260.
Liu, H., Wang, X., Wang, D., Zou, Z., & Liang, Z. (2011). Effect of drought stress on growth and accumulation of active constituents in Salvia miltiorrhiza Bunge. Industrial Crops and Products, 33(1), 84-88.
Liu, Y., Liu, J., Wang, H.-Z., Wu, K.-X., Guo, X.-R., Mu, L.-Q., & Tang, Z.-H. (2020). Comparison of the global metabolic responses to UV-B radiation between two medicinal Astragalus species: an integrated metabolomics strategy. Environmental and Experimental Botany, 104094.
Lum, M., Hanafi, M., Rafii, Y., & Akmar, A. (2014). Effect of drought stress on growth, proline and antioxidant enzyme activities of upland rice. Journal of Animal and Plant Sciences, 24(5), 1487-1493.  
Ma, D., Sun, D., Wang, C., Li, Y., & Guo, T. (2014). Expression of flavonoid biosynthesis genes and accumulation of flavonoid in wheat leaves in response to drought stress. Plant physiology and biochemistry, 80, 60-66.
Mariod, A. A., Ibrahim, R. M., Ismail, M., & Ismail, N. (2009). Antioxidant activity and phenolic content of phenolic rich fractions obtained from black cumin (Nigella sativa) seedcake. Food chemistry, 116(1), 306-312.
Mohammadi, A., Ebrahimzadeh, H., Hadian, J., & Mirmasoumi, M. (2015). Study of the effect of drought stress on some biochemical and physiological parameters of Lippia citriodora HBK. Journal of Plant Research (Iranian Journal of Biology), 28(3), 617-628. (In Persian)
Munné-Bosch, S., & Alegre, L. (2000). Changes in carotenoids, tocopherols and diterpenes during drought and recovery, and the biological significance of chlorophyll loss in Rosmarinus officinalis plants. Planta, 210(6), 925-931.
Nikravesh, M., Kholdebarin, B., Nejadsattari, T., & Najafi, F. (2016). Effect of sodium nitroprusside (SNP) on some physiological parameters in oilseed rape (Brassica napus L.) seedlings under drought stress. Journal of Plant Research (Iranian Journal of Biology), 29(3), 644-658.(In Persian)
Ozkur, O., Ozdemir, F., Bor, M., & Turkan, I. (2009). Physiochemical and antioxidant responses of the perennial xerophyte Capparis ovata Desf. to drought. Environmental and Experimental Botany, 66(3), 487-492.        
Per, T. S., Khan, N. A., Reddy, P. S., Masood, A., Hasanuzzaman, M., Khan, M. I. R., & Anjum, N. A. (2017). Approaches in modulating proline metabolism in plants for salt and drought stress tolerance: Phytohormones, mineral nutrients and transgenics. Plant physiology and biochemistry, 115, 126-140.            
Rahbarian, P., Afsharmanesh, G., & Shirzadi, M. (2010). Effects of drought stress and manure on relative water content and cell membrane stability in dragonhead (Dracocephalum moldavica).      
Saedi, F., Sirousmehr, A., & Javadi, T. (2020). Effect of nano-potassium fertilizer on some morpho-physiological characters of peppermint (Mentha piperita L.) under drought stress. Journal of Plant Research (Iranian Journal of Biology), 33(1), 35-45.
Safikhani, F., Heydari Sharifabad, H., Syadat, A., Sharifi Ashorabadi, A., Syednedjad, M., & Abbaszadeh, B. (2007). The effect of drought stress on percentage and yield of essential oil and physiological characteristics of Dracocephalum moldavica L.  Iranian Journal of Medicinal and Aromatic Plants 23(35), 86-99. (In Persian)
Salehi, A., Tasdighi, H., & Gholamhoseini, M. (2016). Evaluation of proline, chlorophyll, soluble sugar content and uptake of nutrients in the German chamomile (Matricaria chamomilla L.) under drought stress and organic fertilizer treatments. Asian Pacific Journal of Tropical Biomedicine, 6(10), 886-891.            
Sarker, U., & Oba, S. (2018a). Drought stress effects on growth, ROS markers, compatible solutes, phenolics, flavonoids, and antioxidant activity in Amaranthus tricolor. Applied biochemistry and biotechnology, 186(4), 999-1016.
Sarker, U., & Oba, S. (2018b). Drought stress enhances nutritional and bioactive compounds, phenolic acids and antioxidant capacity of Amaranthus leafy vegetable. BMC Plant biology, 18(1), 258.
Saydpour, F., Sayyari, M., & Ghanbari, F. (2015). Effect of glycine betaine on chilling tolerance of cucumber seedlings. Journal of Crops Improvement, 17(1), 53-67. (In Persian)
Sonboli, A., Salehi, P., & Yousefzadi, M. (2004). Antimicrobial activity and chemical composition of the essential oil of Nepeta crispa Willd. from Iran. Zeitschrift für Naturforschung C, 59(9-10), 653-656.           
Stevens, J., Senaratna, T., & Sivasithamparam, K. (2006). Salicylic acid induces salinity tolerance in tomato (Lycopersicon esculentum cv. Roma): associated changes in gas exchange, water relations and membrane stabilisation. Plant Growth Regulation, 49(1), 77-83.  
Taira, S., & Ono, M. (1996). Reduction of astringency in persimmon caused by adhesion of tannins to cell wall fragments. I International Persimmon Symposium 436,          
Trócsányi, E., György, Z., Inotai, K., Szabó, K., Pluhár, Z., Radácsi, P., Malekzadeh, M., & Németh-Zámboriné, E. (2015). Enhanced rosmarinic acid accumulation and rosmarinic acid synthase gene expression under drought stress in thyme (Thymus vulgaris). Planta Medica, 81(16), PM_246.
Turumtay, E. A., İslamoğlu, F., Çavuş, D., Şahin, H., Turumtay, H., & Vanholme, B. (2014). Correlation between phenolic compounds and antioxidant activity of Anzer tea (Thymus praecox Opiz subsp. caucasicus var. caucasicus). Industrial Crops and Products, 52, 687-694.
Xiao, X., Xu, X., & Yang, F. (2008). Adaptive responses to progressive drought stress in two Populus cathayana populations. Silva Fennica, 42(5), 705-719.
Zegaoui, Z., Planchais, S., Cabassa, C., Djebbar, R., Belbachir, O. A., & Carol, P. (2017). Variation in relative water content, proline accumulation and stress gene expression in two cowpea landraces under drought. Journal of plant physiology, 218, 26-34.