Effect of Foliar Application of Amino Acid on the Growth Traits and Phytochemical Composition of Periwinkle (Catharanthus roseus (L.) G. DON) Medicinal Plant in Greenhouse Conditions

Document Type : Research Paper

Authors
Maryam Kaviani 1
Mehrdad Chaichi 2
Hassan Sarikhani 3
1 Assistant Professor, Department of Horticultural Science, Sayyed Jamaleddin Asadabadi University, Asadabad, Iran
2 Assistant Professor, Department of Seed and Plant Improvement Research, Agriculture and Natural Resources Center, Agriculture Research, Education and Extension Organization, Hamedan, Iran.
3 Professor, Department of Horticultural Science, Faculty of Agriculture, Bu-Ali Sina University, Hamedan, Iran
10.22084/ppt.2026.31948.2176
Abstract
Background and Objectives
Periwinkle (Catharanthus roseus (L.) G. DON) is a valuable medicinal plant from the Apocynaceae family, that contains a wide range of secondary metabolites with diverse applications in the pharmaceutical industry. The content of vincristine and vinblastine economically important metabolite is more important than the yield of the plant Researchers have been paying close attention to these alkaloids because of their high price but low presence in the plant. Biostimulants enriched with amino acids have been widely applied to various vegetable crops, demonstrating significant enhancements in nutrient utilization, overall yield, and the quality of the produce.
Nitrogen is the most essential element needed by the plant, which is involved in all the vital processes of plants. Therefore, the use of nitrogen fertilizers for plant production with optimal yield and quantitative and qualitative characteristics is essential. amino acids play a crucial role in enhancing plant nutrient availability and uptake, thereby improving physiological functions and contributing to overall crop quality and yield. Nitrogen availability plays a crucial role in the vegetative growth of plants and has a significant impact on the accumulation of alkaloids in their leaves and roots.
Higher contents of phenols and antioxidants activity in biostimulant amino acid-treated plants can be attributed partially to the indigenous positive effect of amino acids in plant processes. Amino acids are precursors of wide range metabolism secondary metabolites and is tightly link to protein, hormone, and carbohydrate synthesis.
Methodology
The study aimed at investigating the influence of amino acid on the growth traits and phytochemical composition of periwinkle (Catharanthus roseus (L.) G. DON) medicinal plant in Greenhouse Conditions. It utilized a complete randomized design (CRD) with three replications during one consecutive growing season, 2024, in Medicinal herbs garden of Agriculture Research and Training Center, Hamadan. The experiment included five varying level of amino acid including 0, 2.5, 3, 3.5, 4 g-L applied via foliar spraying.
The aerial parts and roots were collected during the blooming period (mid-September). Morphological traits, such as plant height (from the crown to the tip of the main stem) and root length, were measured using a ruler in each replication, and then averaged. The number of flowers and fully grown leaves on each plant was tallied until the end of the growth cycle. To ascertain the fresh and dry weight of the above-ground portions, the plants were severed from the soil surface and weighed using a digital scale. Subsequently, the plants underwent a 48-hour drying process in an oven set at 70 degrees Celsius. Once their weight had reached a stable point, they were weighed again. Also the investigated, physiological and phytochemical traits included leaf chlorophyll, flavonoid, total phenol, antioxidant activity and tryptophan amino acid in the leaves of this plant.
Results
The effect of amino acid on the number of flowers and leaves, root and stem length, fresh and dry weight, chlorophyll a, b and chlorophyll content, phenol, flavonoid, and leaf antioxidant activity (DPPH) was significant (p < 0.01). With increasing levels of amino acid, the amount of all traits increased. The findings demonstrated that the amino acid treatment (3.5 g-L) yielded the highest average count of flowers and leaves, stem and root length, fresh and dry weight of the entire plant, chlorophyll a, antioxidant activity, leaf flavonoids and tryptophan amino acid. The largest quantity of chlorophyll b, total chlorophyll, leaf phenol were associated with the (4 g_L (treatment. Conversely, the control treatment displayed the lowest results. The mean comparison of the treatments showed that the highest number of flowers (8.50) and leaves (63.80), stem length (18.97 cm) and root length (12.98 cm), fresh weight (16.91 g) and total dry weight of the plant (3.36 g), antioxidant activity (48.21% DPPH SC),total flavonoid content (0.561 mg DW-1) and tryptophan amino acid (0.56 mg DW-1) were observed at the level of 3.5 g-L of amino acid.
Conclusion
Overall, applying (3.5 g_L) of amino acid showed the most significant effect on the growth traits and phytochemical composition of periwinkle medicinal plant in Greenhouse Conditions. Based on the results obtained, it can be concluded that the continuous application of amino acid as an organic fertilizer had a positive effect on the growth traits and phytochemical composition of periwinkle. This study's results suggest that applying amino acid biostimulants to leaves enhances periwinkle yields and holds significant promise for cultivating premium periwinkles with enhanced attributes, notably elevated levels of total phenolics, flavonoids, and antioxidant capacity.
Keywords
Antioxidant
Flavonoid
Foliar application
Photosynthetic pigments
Subjects
Abd El-Aziz, N. G., Mahgoub, M. H. and Mazher, A. A. (2009). Physiological effect of phenylalanine and tryptophan on the growth and chemical constituents of Antirrhinum majus plants. Ozean Journal of Applied Science, 2(4): 399-407.
Abdelkader, M., Voronina, L., Baratova, L., Shelepova, O., Zargar, M., Puchkov, M. and Arinov, B. (2023). Biostimulants-based amino acids augment physio-biochemical responses and promote salinity tolerance of lettuce plants (Lactuca sativa L.). Horticulturae, 9(7): 807. https://doi.org/10.3390/horticulturae9070807
Al-Karaki, G. N. and Othman, Y. (2023). Effect of foliar application of amino acid biostimulants on growth, macronutrient, total phenol contents and antioxidant activity of soilless grown lettuce cultivars. South African Journal of Botany, 154, 225-231. https://doi.org/10.1016/j.sajb.2023.01.034
Almagro, L., Fernández-Pérez, F. and Pedreño, M. A. (2015). Indole alkaloids from Catharanthus roseus: bioproduction and their effect on human health. Molecules, 20(2): 2973-3000. https://doi.org/10.3390/molecules20022973
Bidaki, S. and Tehranifar, A. (2018). Post-harvest shelf-life extension of fruits of two strawberry (Fragaria× ananassa Duch.) cultivars with amino acids application in soilless culture system. Journal of Soil and Plant Interactions-Isfahan University of Technology, 9(2): 1-10. https://doi.org/10.29252/ejgcst.9.2.1
Chang, C. C., Yang, M. H., Wen, H. M. and Chern, J. C. (2002). Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of Food and Drug Analysis. 10(3). https://doi.org/10.38212/2224-6614.2748
El-Sayed, M. and Verpoorte, R.(2007). Catharanthus terpenoid indole alkaloids: biosynthesis and regulation. Reviews; 6: 277-305.
Estefan, G., Sommer, R., and Ryan, J., (2013). Methods of soil, plant, and water analysis. A manual for the West Asia and North Africa region. 3(2). https://doi.org/10.1186/s42269-019-0067-0
Haghparvar, F. R., Hashemabadi, D. and Kaviani, B. (2023). The effect of foliar application of amino acids on some nutritional properties, antioxidant capacity and some other physiologic parameters of African marigold (Tagetes erecta L.), Taishan ‘Yellow’and ‘Orange’. Acta Scientiarum Polonorum Hortorum Cultus, 22(1): 107-124. https://doi.org/10.24326/asphc.2023.4580
Hatano, T., Kagawa, H., Yasuhara, T. and Okuda, T. (1988). Two new flavonoids and other constituents in licorice root: their relative astringency and radical scavenging effects. Chemical and Pharmaceutical Bulletin. 36(6): 2090-2097. https://doi.org/10.1248/cpb.36.2090
Henderson, B. C., Sanderson, J. M. and Fowles, A. (2025). A review of the foliar application of individual amino acids as biostimulants in plants. Discover Agriculture, 3(1): 69. https://doi.org/10.1007/s44279-025- 00222-7.
Hosokawa, S., Morinishi, T., Ohara, K., Yamaguchi, K., Tada, S. and Tokuhara, Y. (2023). A spectrophotometric method for the determination of tryptophan following oxidation by the addition of sodium hypochlorite pentahydrate PLoS ONE, 18(1), e0279547. https://doi.org/10.1371/journal. pone.0279547
Hu, W., Coomer, TD., Loka, DA., Oosterhuis, DM. and Zhou, Z.(2017). Potassium deficiency affects the carbon-nitrogen balance in cotton leaves. Plant physiological chemistry; 115:408-417.
Jahanbani, S., Mumivand, H., Zahedi, B. and Argento, S. (2024). Foliar Application of Urea and Amino Acids Regulates Growth, Photosynthesis, Pigments, Antioxidant Activity, and the Essential Oil Content and Composition of Basil (Ocimum basilicum L.). Agronomy, 14(12): 2950. https://doi.org/10.3390/agronomy14122950
Jalili Moghadam, Z. J., Kalantari, S., Estahbanati, A. H. and Garrido, D. V. (2019). Effect of Amino acids and Pepton foliar applications on quality and storage life of pomegranate fruit cv. Malas-e-Save. Iranian Journal of Horticultural Science, 50(3), 527-537. https://doi.org/ 10.22059/ijhs.2018.261060.1465
Khalifa, Y. A., El-Naem, A., Gamal, F. and Mahmoud, M. A. (2020). Effect of tryptophan and ascorbic acid on yield and some chemical constituents of lupine (Lupines termis L.) plants. Egyptian Journal of Agronomy, 42(1), 47-61.
Khan, A. S., Ahmad, B., Jaskani, M. J., Ahmad, R. and Malik, A. U. (2012). Foliar application of mixture of amino acids and seaweed (Ascophylum nodosum) extract improve growth and physicochemical properties of grapes. International Journal of Agriculture and Biology, 14(3): 383-388.
Khazaei, Z., and Sayari, M. (2015). Effect of 5 Aminololic acid leaf application on growth and some physiologic factors and sweet pepper (Capsicum annuum L.) function in drought stress conditions. Journal of Plant Research (Iranian Journal of Biology), 28 (5): 952-961. https://doi.org/10.22077/jsr.2021.4762.1172
Lambers, H., Chapin III, F. S. and Pons, T. L. (2008). Plant physiological ecology. Springer Science and Business Media.
Lichtenthaler, H. K. and Buschmann, C. (2001). Chlorophylls and carotenoids: Measurement and characterization by UV‐VIS spectroscopy. Current Protocols in Food Analytical Chemistry. 1(1): F4-3. https://doi.org/10.1002/0471142913.faf0403s01
Moore, MR., Williams, and DJ., Chia J.(1967). Factor affecting concentrations of dimethylated indole alkylamines in phalaris tuberosa L. Journal Biologist Science.; 20:1131-40.
Noviyanti, R., Sari, R. L. K., Kristanti, A. N., Yachya, A. and Manuhara, Y. S. W. (2017). Biomass and flavonoid production of Gynura procumbens adventitious roots induced by sucrose, phenylalanine and tyrosine. Bioscience Research, 14(4): 934-941. https://www.researchgate.net/publication/322558300
Oraghi Ardebili, Z., Laden Moghadam, AS. R. and Moradi, M. (2011). Effect of Leaf Seedling on Selenium and Amino Acid on Parsley. Garmsar Azad University. Garmsar master’s Thesis.
Oudin, A., Courtois, M., Rideau, M. and Clastre M.(2007). The pathway in Catharanthus roseus alkaloid biosynthesis. Phytochemistry Reviews; 6 (2): 259-76.
Pirvash, A., Sheikh-Mohseni, M. and Nejadhabibvash, F. (2021). Morphological characteristics and total phenol concentration in hydroalcoholic extract of Echinacea purpurea L. influenced by different nitrogen sources. Iranian Journal of Medicinal and Aromatic Plants Research, 37(3): 457-474. https://doi: 10.22092/ijmapr.2021.353236.2936
Rabiei, V., Akbari, A. and Barzegar, T. (2024). The effect of low temperature on fruit yield, quality and antioxidant properties of physalis under foliar spray of amino acids and selenium. Journal of Plant Production Research, 31(3): 43-65. https://doi: 10.22069/jopp.2023.21517.3060
Rady, M. R. (2019). Plant biotechnology and milk thistle. In Plant Biotechnology and Medicinal Plants: Periwinkle, Milk Thistle and Foxglove Cham: Springer International Publishing. 97-147.
Ramzan, T., Shahbaz, M., Maqsood, M. F., Zulfiqar, U., Saman, R. U., Lili, N. and Haider, F. U. (2023). Phenylalanine supply alleviates the drought stress in mustard (Brassica campestris) by modulating plant growth, photosynthesis, and antioxidant defense system. Plant Physiology and Biochemistry, 201, 107828:https://doi.org/10.1016/j.plaphy.2023.107828
Rani, V., Kumar, V., Meena, R., Jain, S. K. and Birla, D. (2023). Impact of Nitrogen Levels on the Growth and Medicinal Properties of Periwinkle (Catharanthus roseus) in an Inceptisol of Varanasi, India. International Journal of Plant and Soil Science. 35(19): 954-962. https://doi.org/10.9734/IJPSS/2023/v35i193630
Reham, M. S., Khattab, M. E., Ahmed, S. S. and Kandil, M. A. M. (2016). Influence of foliar spray with phenylalanine and nickel on growth, yield quality and chemical composition of genoveser basil plant. African Journal of Agricultural Research, 11(16): 1398-1410. https://doi.org/10.5897/AJAR2015.10699
Rezakhani, A. and Hadi, M. R. H. S. (2017). Effect of manure and foliar application of amino acids on growth characteristics, seed yield and essential oil of coriander (Coriandrum sativum L.). Iranian Journal of Field Crop Science, 48(3): 777-786.
Roy, A. A review on the alkaloids an important therapeutic compound from plants.(2017). International Journal of Plant and  biotechnology.; 3:1-9.
Shafie, F., Bayat, H., Aminifard, M. H. and Daghighi, S. (2021). Biostimulant effects of seaweed extract and amino acids on growth, antioxidants, and nutrient content of yarrow (Achillea millefolium L.) in the field and greenhouse conditions. Communications in Soil Science and Plant Analysis, 52(9): 964-975. https://doi.org/10.1080/00103624.2021.1872596
Shekari, G. and Javanmardi, J. (2017). Application of cysteine, methionine and amino acid containing fertilizers to replace urea: The effects on yield and quality of Broccoli. Advances in Crop Science and Technology, 5(3): 1-4. https://doi.org/10.4172/2329-8863.1000283
Singleton, V.L., Orthofer, R. and Lamuela-Raventos, R.M. (1999). Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods in Enzymology. 299: 152-178.
https://doi.org/10.1016/S0076-6879(99)99017-1
Stiegler, J. C., Richardson, M. D., Karcher, D. E., Roberts, T. L. and Norman, R. J. (2013). Foliar absorption of various inorganic and organic nitrogen sources by creeping bentgrass. Crop Science, 53(3): 1148-1152. https://doi.org/10.2135/cropsci2012.08.0511
Sun, W. and Shahrajabian, M. H. (2023). Therapeutic potential of phenolic compounds in medicinal plants—Natural health products for human health. Molecules, 28(4): 1845. https://doi.org/10.3390/molecules28041845
Sun, W., Shahrajabian, M. H., Kuang, Y., and Wang, N. (2024). Amino Acids Biostimulants and Protein Hydrolysates in Agricultural Sciences. Plants, 13(2):210. https://doi.org/10.3390/plants13020210
Taha, HS., El- Bahr, MK. and Seif El Nasr, M. M. (2009). In vitro studies on egyptian Catharanthus roseus (L.) G.Don. IV: manipulation of some amino acids as precursors for enhanced of indole alkaloids production in suspension cultures. Aust. Journal Basic Application of Science, 3: 3137-44.
Tarasevičienė, Ž., Velička, A. and Paulauskienė, A. (2021). Impact of foliar application of amino acids on total phenols, phenolic acids content of different mints varieties under the field condition. Plants, 10(3): 599. https://doi.org/10.3390/plants10030599
Teixeira, W. F., Fagan, E. B., Soares, L. H., Soares, J. N., Reichardt, K. and Neto, D. D. (2018). Seed and foliar application of amino acids improve variables of nitrogen metabolism and productivity in soybean crop. Frontiers in Plant Science, 9, 396. https://doi.org/10.3389/fpls.2018.00396
Teixeira, W. F., Fagan, E. B., Soares, L. H., Umburanas, R. C., Reichardt, K. and Neto, D. D. (2017). Foliar and seed application of amino acids affects the antioxidant metabolism of the soybean crop. Frontiers in plant science, 8, 327. https://doi.org/10.3389/fpls.2017.00327
Tzin, V. and Galili, G. (2010). The biosynthetic pathways for shikimate and aromatic amino acids in Arabidopsis thaliana. The Arabidopsis book/American Society of Plant Biologists, 8p.
Wassel, A. H. M., Gobara, A. A., Ibrahiem, H. I. M. and Shaaban-Mai, M. (2015). Response of wonderful pomegranate trees to foliar application of amino acids, vitamins B and silicon. World Rural Observations, 7(3): 91-95.
Whitmer, S., Van Der Heijden, R. and Verpoorte, R. (2002). Effect of precursor feeding on alkaloid accumulation by a tryptophan decarboxylase over-expressing transgenic cell line T22 of Catharanthus roseus. Journal of Plant and  biotechnology.; 96: 193-203.
Zhou, W., Zheng, W., Lv, H., Wang, Q., Liang, B. and Li, J. (2022). Foliar application of pig blood-derived protein hydrolysates improves antioxidant activities in lettuce by regulating phenolic biosynthesis without compromising yield production. Scientia Horticulturae, 291:110602. https://doi.org/10.1016/j.scienta.2021.110602
Volume 18, Issue 1
September 2026
Pages 1-13