In collaboration with Scientific Association of Iranian Medicinal Plants

Document Type : Research Paper

Authors

1 Department of Horticulture, Faculty of Agriculture, University of Zanjan, Iran

2 Department of Horticultural science, Faculty of Agriculture, Universit of Zanjan, Iran

3 Faculty of Agriculture- Horticulture University of Zanjan, Iran

4 Department of Chemical Engineering, Faculty of Engineering, University of Zanjan, Zanjan, Iran

Abstract

Background and objectives: Digitalis purpurea L., the most well-known species of the Digitalis genus, is a biennial plant containing cardiac glycosides (0.3 to 0.4%) in its leaves. These glycosides are extracted and used as natural medicines for treating heart diseases, with no chemical substitutes available, making them widely utilized annually. In recent years, using amino acids and growth regulators as alternatives to chemical compounds has gained popularity for enhancing secondary metabolites, improving the quality and quantity of agricultural products, and promoting sustainable agriculture. This study investigates the biochemical and growth responses of D. purpurea to foliar applications of glutamic acid and benzyladenine under greenhouse conditions to evaluate their effects on morphophysiological characteristics, yield improvement, and their potential as environmentally friendly alternatives to chemical fertilizers.
Methodology: A completely randomized design with three replications was conducted in the research greenhouse of the University of Zanjan to evaluate the effects of foliar applications of benzyladenine (0.5 and 1 mM) and glutamic acid (1 and 2 mM), along with a control treatment (distilled water). Seeds were initially planted in a seedling tray containing cocopeat and peat moss. At the four-leaf stage, three seedlings were transplanted into plastic pots filled with a culture medium of field soil, cocopeat, and perlite (1:1:3 ratio). Greenhouse conditions were maintained at average day and night temperatures of 25°C and 18°C, respectively, with 80% relative humidity. Seedlings were irrigated weekly with 100 mL of complete Hoagland solution. Foliar spraying with the treatments began after seedling establishment in late June and was repeated four times at 10-day intervals. Leaf samples were collected 10 days after the final application for laboratory analysis. Data were analyzed using SAS software (version 9), and means were compared using Duncan’s multiple range test at a 5% probability level.
Results: The application of 1 mM benzyladenine resulted in the highest total chlorophyll content (1.95 mg/g fresh weight), antioxidant activity (36.02%), and peroxidase enzyme activity (1.14 U/mg protein per minute). The maximum total carotenoid content (0.14 mg/g fresh weight), total phenol (12.52 mg gallic acid/g fresh weight), total flavonoid (2.86 mg quercetin/g fresh weight), and nitrogen content (1.46%) were achieved with one mM glutamic acid. Additionally, one mM glutamic acid application led to the highest catalase enzyme activity (4.09 U/mg protein per minute), fresh weight (123.51 g), and dry weight (47.38 g).
Conclusion: The findings suggest that applying glutamic acid and benzyladenine at varying levels can significantly enhance the biochemical and growth characteristics of Digitalis purpurea L.

Keywords

Main Subjects

- Abdel-Aziz, N.G., Taha Lobna, S. and Ibrahim Soad, M.M., 2009. Some studies on the effect of putrescine, ascorbic acid and thiamine on growth, flowering and some chemical constituents of Gladiolus plants at Nubaria. Ozean Journal of Applied Sciences, 2(2): 169-179.
- Abdel-Aziz, N., El-Quesni F.E.M. and Farahat, M.M., 2007. Response of vegetative growth and some chemical constituents of Syngonium podophyllum L. to foliar application of thiamine, ascorbic acid and kinetin at Nubaria. World Journal of Agricultural Science, 3(3): 301-305.
- Abou Rayya, S.M., Thanaa, SH.M. and Nabila, E.K., 2015. Photosynthetic pigments and fruit quality of Manzanillo olive as affected by 6-benzyl adenine and studying the chemical constituents in leaves using Fourier transform infrared spectroscopy technique. International Journal of ChemTech Research, 8(6): 514-522.
- Abshahi, M., Zarei, H., Ghasemnezhad, A. and Aghdasi, M., 2014. Changes in chlorophyll contents, external quality characters, dry weight, and vase life of strelitzia (Strelitzia spp.) leaves affected by different preservative solutions. Journal of Plant Physiology and Breeding, 4(2): 1-8.
- Amini Fard, M., Gholami, M., Bayat, H. and Moradi Nejad., 2017. Investigating the effect of fulvic acid and amino acid fertilizers on phenolic compounds, flavonoids, antioxidant activity and photosynthetic pigments of the medicinal plant (Coriandrum sativum L.). Ecophytochemistry Quarterly of Medicinal Plants, 25(7): 25-39. (In Persian)
- Aremu, A.O., Fawole, O.A., Makunga, N.P., Masondo, N.A., Moyo, M., Buthelezi, N.M.D., Amoo, S.O., Spichal, L. and Dolezal, k., 2020. Applications of cytokinins in horticultural fruit crops. Trends and future prospects. Biomolecules, 10(9): 1222.
- Arnon, A.N., 1967. Method of extraction of chlorophyll in the plants. Agronomy Journal, 23:112-121.
- Ashrafi, S., 2013. The effect of foliar spraying of urea and some amino acids on the morphological, physiological and metabolic characteristics of Tanacetum balsamita L. Master's thesis in the field of horticultural sciences, Urmia University, Iran. (In Persian)
- Azza, A.M., Zaghloul, S.M., Mahmoud, S.A. and Hanan, S., 2011. Stimulatory effect of kinetin, ascorbic acid and glutamic acid on growth and chemical onstituents of Codiaeum variegatum L. plants. American-Eurasian Journal of Agricultural and Environmental Sciences, 10(3): 318-323.
- Brand-Williams, W., Cuvelier, M.E. and Berset, C., 1995. Use of a free radical method to evaluate antioxidant activity. LWT- Food Science and Technology, 28(1): 25-30.
- Chang, C., Yang, M., Wen, H. and Chern, J., 2003. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of Food and Drug Analaysis, 10(3): 178-182.
- Chen, G. and Gao, X., 2002. Effect of partial replacement of nitrate by amino acid and urea on nitrate content of nonheading Chinese cabbage and lettuce in hydroponics (Chinese). Scientia Agricultura Sinica, 35(3): 187–191.
- Dhindsa, R.S., Plumb‐Dhindsa, P. and. Thorpe, T.A., 1981. Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. Journal of Experimental Botany, 32(126): 93–101.
- Duck, M.W., Gregg, B.M., Fernandez, R.T., Royal, D.H. and Cardoso, F.F., 2004. Height control of Picea spp. and Chamaecyparis lawsoniana with uniconazole and 6-benzyladenine. Journal of Environmental Horticulture, 22(3): 165–169.
- Emami, A., 1996. Methods of Plant Decomposition. Agricultural Research, Education and Extension Organization, Soil and Water Research Institute, Tehran, Iran, 126p. (In Persian)
- Fallahi, H.R., Amini Fard, M.H. and Jorkesh, A., 2018. Effects of thiamine spraying on bio-chemical and morphological traits of basil plants under greenhouse conditions. Journal of Horticulther and Postharvest Research, 1(1): 27-36.
- Farouk, S., Youssef, S.A. and Ali, A., 2012. Exploitation of bio stimulants and vitamins as an alternative strategy to control early blight of tomato plants. Asian Journal of Plant Science, 11(1): 36-43.
- Faten, S.A., Shaheen, A.M. Ahmed, A.A. and Mahmoud, A.R., 2010. Effect of foliar application of amino acids as antioxidants on growth, yield and characteristics of Squash. Agriculture and Biological Science, 5(6): 583- 588.
- Garcia, L.A., Madrid, R., Gimeno, V., Rodriguez-Ortega, W.M., Nicolas, N. and Garcia-Sanchez, F., 2011. The effects of amino acids fertilization incorporated to the nutrient solution on mineral composition and growth in tomato seedlings. Spanish Journal of Agricultural Research, 9(3): 852- 861.
- García-Ramírez, M.G. Gonzáles, E.Q. Mendoza, M. . Seijo, M.L. Cárdenas, L.J. Moreno-Bermúdez, O. and. Ribalta, H., 2014. Effect of BA treatments on morphology and physiology of proliferated shoots of Bambusa vulgaris Schrad. Ex Wendl in temporary immersion. American Journal of Plant Sciences, 5(2): 205-211.
- Ghazi Manas, M., Banj Shafiee, S., Haj Seyed Hadi, M.R. and Darzi, M.T., 2013. Effects of vermicompost and nitrogen on quantitative and qualitative yeild of chamomile (Matricaria chamomilla L.). Iranian Journal of Medicinal and Aromatic Plants research, 29(2): 269-280.
- Gorni, P.H. and Pacheco, A.C., 2016. Growth promotion and elicitor activity of salicylic acid in Achillea millefolium L. African Journal of Biotechnology, 15(16): 657-665.
- Hanaa, H., Hussein, A., Game, M.M., and El-Baroty, S., 2008. Algal extracts improve antioxidant defense abilities and salt tolerance of wheat plant irrigated with sea water. International Journal of Irrigation and Water Management, 7(11): 2812-2832.
- Hasnaa, S., Ayad, M. Karima, M. and EL-Din, G., 2011. Effect of atonik and benzyladenine on growth and some biochemical constituents of lupine plant (Lupinus termis L.). American-Eurasian Journal of Agriculture and Environmental Science, 10 (4): 519-524.
- Hassan, E.H. and Ali, E.F., 2010. Response of Coriandrum sativum L. plant to cutting as well as glycine and salisylic acid treatments. Minia Journal of Agricultural Research and Development, 30(1): 15-29.
- Hazrati Yadekori, S. and Tahmasebi Sarvestani, Z., 2012. Effects of different nitrogen fertilizer levels and hormone benzyl adenine (BA) on growth and ramet production of Aloe vera L. Iranian Journal of Medicinal and Aromatic Plants research, 28(2): 210-223. (In Persian)
- Hendawy, S.F. and Ezz-Din, A.A., 2010. Growth and yield of Foeniculum vulgare var. azoricum as influenced by some vitamins and amino acids. Journal of Applied Sciences, 3(1): 113-123.
- Hoagland, D.R. and Arnon, D.J., 1950. The water culture method for growing plants without soil. 2nd Ed. California Agricultural Experimental Station, Berkeley, 32p.
- Hussein, A.A., 2005. Biochemical and Physiological Studies on some Alkaloids Ph.D. Thesis, Faculty of Agriculture Cairo University.
- Jahani., R., Hosni, A. and Samadi, A., 2016. The effect of spraying urea, aspartic acid and glutamic acid on the hypertensive, physiological and biochemical characteristics of Agastache foeniculum. Journal of soil use research. 5(2): 95-107.
- Karima, A., Gamal El-Din, K.M. and Abdel-Wahed, M.S.A., 2005. Effect of some amino acids on growth and essential oil content of chamomile plant. International Journal of Agriculture and Biology, 7(3): 376-380.
- Khattab, M., Shehata, A., Abou El -Saadate, E. and Al-Hasni, Kh., 2016. Effect of glycine, methionine and tryptophan on the vegetative growth, flowering and corms production of gladiolus plant. Alexandria Science Exchange Journal, 37(4): 647-659.
- Lopez, M.L., Peralta-Videa, J.R., Benitez, T. and Gardea-Torresdey, J.L., 2005. Enhancement of lead uptake by alfalfa (Medicago sativa) using EDTA and a plant growth promoter. Chemosphere, 61(4): 595-598.
- Lukaszewska, A., Monika, P. and Karol, C.H., 2008. Effect of drought and benzyl adenine on scarlet salvia (Salvia splendens Sello) and geranium (Pelargonium hortorum L.). Annals of Warsaw University of Life Sciences–SGGW 29(6): 45–52.
- Meda, A., Lamien, CE. Romito, M. Millogo, J. and Nacoulma, O.G., 2005. Determination of the total phenolic, flavonoid and pralin contents in Burkina Fasan honey, as well as their scavenging activity. Food Chemistry, 91(1): 571-577.
- Michalak, A., 2006. Phenolic compounds and their antioxidant activity in plants growing under heavy metal stress. Polish Journal of Environmental Studies, 15(4): 523- 530.
- Natesan, R., Kandasamy, S. Thiyageshwari, S. and Boopathy, P.M., 2007. Influence of lignite humic acid on the micronutrient availability and yield of blackgram in an alfisol. Scientific World Journal, 7(11):1198-1206.
- Nguyen, P.H.M., Kwee, E.M. and Niemeyer, E.D., 2010. Potassium rate alters the antioxidant capacity and phenolic concentration of basil (Ocimum basilicum L.) leaves. Food Chemistry, 123(4): 1235-1241.
- Nisar, B., Sultan, A. and Rubab, S.L., 2018. Comparison of medicinally important natural products versus synthetic drugs-a short commentary. Natural Products Chemistry and Research, 6(2): 1-2.
- Olmstead, R.G.; Pamphilis, C.W., Wolfe, A.D., Young, N.D., Elisons, W.J., and Reeves, P.A., 2001. Disintegration of the Scrophulariaceae. American Journal of Botany, 88 (2): 348–361.
- Omidbeigi, R., 2007. Production and Processing of Medicinal Plants. Vol. II, Behnashr, Mashhad, 442p. (In Persian)
- Oraghi Ardebili, Z., Ladan Moghadam, A.R. Oraghi Ardebili, N. and Pashaie, A.R., 2012. The induced physiological changes by foliar application of amino acids in Aloe vera L. plants. Plant Omics Journal, 5(3): 279-284.
- Paciorek, T., Zazimalova, E., Ruthardt, N., Petrasek, J., Stierhof, Y.D., Kleine-Vehn, J., Morris, D.A., Emans, N., Jurgens, G. and Geldner, N., 2005. Auxin inhibits endocytosis and promotes its own efflux from cells. Nature, 435: 1251–1256.
- Piotrowska, A.; and Czerpak, R., 2009. Cellular response of light/dark-grown green alga Chlorella vulgaris Beijerinck (Chlorophyceae) to exogenous adenine and phenylurea-type cytokinins. Acta Physiologiae Plantarum, 31(3): 573–585.
- Ramtin, A., Kalatejari, S., Naderi, R. and Matinizadeh, M., 2016. Effect of benzyl adenine and salicylic acid on biochemical traits of two cultivars of carnation. Journal of Experimental Biology and Agricultural Sciences, 4(4): 427-434.
- Reda, F., Baroty, G.S.A., Talaat, I.M., Abdel-Rahim, I.A. and Ayad, H.S., 2007. Effect of some growth regulators and vitamins on essential oil, phenolic content and activity of oxidoreductase enzymes of Thymus vulgaris L. World Journal of Agricultural Sciences, 3(5): 630-638.
- Shehata, S.M., Azem, S. and El-Yazied, A., 2011. Effect of Foliar Spraying with Amino Acids and Seaweed Extract on Growth Chemical Constitutes, Yield and its Quality of Celeriac Plant. European Journal of Scientific Research, 58(2): 257-265.
- Taiz, L. and Zeiger, E., 2006. Plant Physiology. 4th ed. Sinauer Associates Inc. Publishers, Massachusetts, US. 623p.
- Tajik, N. and Danaee, E., 2016. Study the effect of spraying pre-harvest glotamin, arginine and phenylalanine on some physicochemical and enzymatic traits and longevity gerbera jamesonii flower cv. Sorbet Cellular and Molecular Plant Biology Journal, 8(3): 4-12.
- Verma, S.K., Gantait, S. Jeong, B.R. and Hwang, S.J., 2018. Enhanced growth and cardenolides production in Digitalis purpurea under the influence of different LED exposures in the plant factory. Journal of Scientific Reports, 8(1): 1-12.
- Wang, S., Wang, H.B., Wang, X.D., Shi, X.B., Wang, B.L., Zheng, X.C. and Liu, F.Z., 2015. Effects of selenium and 6-BA on leaf senescence and active oxygen metabolism in grape. Journal of fruit science, 32(11): 206–214.