The Effect of Biofertilizer Application on the Improvement of Pigments, Physiological Traits, and Yield of Cowpea (Vigna unguiculata(

بوم شناسی کشاورزی

Article 6, Volume 17, Issue 2 - Serial Number 64, June 2025, Pages 281-302   

Document Type: Research Article

DOI: 10.22067/agry.2025.89695.1213

Authors

Reza Sabaghian¹; Mehdi Baradaran firouzabadi^{* 1}; Mostafa Haydari¹; farzane bahadori²; Safiye Arab¹

¹Department of Agriculture, Faculty of Agriculture, Shahrood University of Technology, Shahrood, Iran

²Research and Education Center for Agriculture and Natural Resources of Semnan Province, Semnan, Iran.

Abstract

Introduction
Cowpea (Vigna unguiculata) an annual legume, originated in Iran. It is one of the main sources of protein, amino acids, vitamins, minerals, and energy, and therefore is important for a well-balanced diet with good nutritional value. In recent years, the application of organic fertilizers has increased tremendously in agricultural production. Humic compounds such as HA is one of the organic-mineral fertilizers, which is commonly used for the production of crops. In order to increase the quantitative and qualitative traits of plants, the extract of some plants, such as green tea extract and coffee extract, can be used.
 
Materials and Methods
In order to investigate the effect of biofertilizers, including humic acid, green tea extract, and coffee extract, on the improvement of pigments and physiological traits of cowpea, an experiment was designed and implemented in 2023 at a private farm in Semnan city. The experiment was a factorial format based on a randomized complete block design with three replications. The test treatments include seed priming at five levels (control, distilled water, humic acid (750 mg/liter), green tea extract (10% concentration) and coffee extract (10% concentration)) and foliar spraying at four levels (control, humic acid (750 mg/liter), green tea extract (concentration 10%) and coffee extract (concentration 10%). Pretreatment of seeds with desired concentrations was done for six hours, following the principles of seed aeration. Foliar spraying was done at the beginning of flowering. Seeds were sown on June 25 (as the second planting). Before planting, all the seeds were disinfected with benomyl and then in each plot, four rows of five meters were planted with a distance of 25 cm between the rows and 8 cm within the row at a depth of 5 cm.
Results and Discussion
The results showed that pretreatment of seeds with humic acid, green tea extract and coffee extract led to an increase in the traits of relative leaf water content, membrane stability index, leaf surface index, photosynthetic pigments, protein yield, and grain yield. In plants grown from pre-treated seeds, the amount of malondialdehyde and soluble carbohydrates in leaves decreased to a significant level. Foliar spraying with these substances led to the improvement of leaf surface index, the amount of photosynthetic pigments, protein yield, and seed yield. Application of foliar spraying also reduced the amount of malondialdehyde and soluble carbohydrates in leaves. Finally, within the scope of the research, the use of humic acid at the rate of 750 mg/liter and green tea at a concentration of 10% is recommended as seed pre-treatment and foliar spraying in the field. More research is needed on the recommendation of seed pretreatment and coffee extract foliar spraying. When seed yield was considered as a dependent trait, the traits of leaf surface index (0.6076), carotenoid (0.7265), membrane stability index (0.6501), leaf soluble carbohydrates (-0.6678), and Malondialdehyde (0.6096) entered the model as main variables. The results of this research showed that carotenoid (0.3764) and membrane stability index (0.2950) had the most positive direct effect on grain yield.
Conclusion
The results of this research showed that the photosynthetic pigments, protein yield, and grain yield of cowpea increased to a significant level by using humic acid, green tea extract, and coffee extract as seed pretreatment and foliar spraying. Seed protein percentage, yield components, and grain yield of cowpea increased to a significant level with the application of humic acid, green tea extract, and coffee extract as seed pretreatment and foliar spraying. Finally, within the scope of this research, the use of humic acid at the rate of 750 mg/L and green tea at a concentration of 10% is recommended as seed pretreatment and foliar spraying in the field. More research is needed on the recommendation of seed pretreatment and foliar spraying of coffee extract.



 

 




 
 

Keywords

Antioxidant; Biological fertilizers; Foliar spraying; Pretreatment

References

1. Abada, M.A. (2014). A comparative study for the effect of green tea extract and some antioxidants on Thompson Seedless grapevines. International Journal of Plant & Soil Science, 3(10), 1333-1342.‏ https://doi.org/9734/ljpss/2014/8611
2. Abdulameer, O.Q., & Ahmed, S.A. (2019). Role of humic acid in improving growth characters of corn under water stress. The Iraqi Journal of Agricultural Science, 50(1), 420-430.‏
3. Aggag, A.M., Alzoheiry, A.M., & Abdallah, A.E. (2015). Effect of kaolin and fulvic acid anti-transpirants on tomato plants grown under different water regimes. Alexandria Sciene Exchange Journal, 36, 2-15. https://doi.org/10.21608/asejaiqjsae.2015.2875
4. Ahmed, F.F., Kamel, M.K., & Ibrahim, H.I.M. (2014). The synergistic effects of using plant extracts and salicylic acid on yield and fruit quality of Keitte mango trees. Stem Cell, 5(2), 30-39.
5. Alipour, H., Razavinasab, A., & Hosseinifard, S.J. (2023). The effect of humic acid application on growth characteristics, quality and yield of pistachios. Journal of Soil Management and Sustainable Production, 12(4), 117-133. (In Persian with English abstract). https://doi.org/22069/ejsms.2023.20659.2077  
6. Arnaout, S.M., Kamel, N.H., El-Mosallamy, H.M.H., & Boghdady, M.S. (2019). Effect of unihumic and aminomore on cowpea plants (Vigna unguiculata) grown under drought stress conditions. Zagazig Journal of Agricultural Research, 46(4), 999-1019. https://doi.org/10.21608/zjar.2019.47053
7. Balasundram, N., Sundram, K., & Samman, S. (2006). Phenolic compounds in plants and agri-industrial by-products: Antioxidant activity, occurrence, and potential uses, Food Chemistry, 103, 1288-1296. https://doi.org/org/10.1016/j.foodchem.2005.07.042
8. Balzano, M., Loizzo, M.R., Tundis, R., Lucci, P., Nunez, O., Fiorini, D., Giardinieri, A., & Pacetti, D. (2020). Spent espresso coffee grounds as a source of anti-proliferative and antioxidant compounds. Innovative Food Science & Emerging Technologies, 59, 102-125. https://doi.org/10.1016/j.ifset.2019.102254
9. Bidyabhusan, B., Kangujam, B., Barkha, S., & Arambam, A. (2024). Effect of humic acid on growth, yield and soil properties in rice: A review. International Journal of Plant & Soil Science, 36(6), 26-35. https://doi.org/9734/ijpss/2024/v36i64603
10. Bili, C. (2015). Mechanism of silicon alleviation of drought stress in tomato (Solanum lycopersicum). Shandong Agricultural University, 214 pp.
11. Castaldo, L., Toriello, M., Izzo, L., Sessa, R., Lombardi, S., Trombetti, S., & Grosso, M. (2022). Effect of different coffee brews on tryptophan metabolite-induced cytotoxicity in HT-29 human colon cancer cells. Antioxidants, 11(12), 24-38. https://doi.org//10.3390/antiox11122458
12. Cavusoglu, D., Macar, O., Kalefetoglu Macar, T., Cavusoglu, K., & Yalcın, E. (2022). Mitigative effect of green tea extract against mercury (II) chloride toxicity in Allium cepa model. Environmental Science and Pollution Research, 29(19), 27862-27874. https://doi.org//10.1007/s11356-021-17781-z
13. Cordeiro, F.C., Santa-Catarina, C., Silveira, V., & Souza, S.R. (2011). Humic acid effect on catalase activity and the generation of reactive oxygen species in corn (Zea mays). Bioscience Biotechnology and Biochemistry, 75, 70–74. https://doi.org/10.1271/bbb.100553
14. Dahmarde, M., Mirbahaodin, M., & Khomri, A. (2017). The effect of biofertilizer application on the quantitative and qualitative characteristics of cowpea under drought stress conditions. Environmental Stresses in Agricultural Sciences, 11(1), 23-33. (In Persian with English abstract).
15. Daylami, S., & Mojaddam, M. (2018). Studying the effect of application time of humic acid foliar spraying on qualitative and quantitative characteristics of cowpea (Vigna unguiculata) under different irrigation regimes. Environmental Stresses in Crop Sciences, 12(1), 85-94. (In Persian with English abstract). http://dx.doi.org/10.22077/escs.2018.1056.1209
16. Du, Z., & BramLey, W.J. (1992). Modified thiobarbituric acid assay for measuring lipid oxidation in sugar-rich plant tissue extracts. Journal Agricultural and Food Chemistry, 40(9), 1566-1570. https://doi.org/10.1021/jf00021a018
17. Eryigit, T., & Husamalddin, A.H. (2023). Effects of different humic acid doses on yield and quality properties of corn (Zea mays) in Iraq-Sulaymaniyah conditions. Journal of the Institute of Science and Technology, 13(2), 1377-1393.‏ https://doi.org/10.21597/jist.1241745
18. Fathi, A., Farnia, A., & Maleki, A. (2016). Effect of nitrogen and phosphorus biological fertilizers on vegetative characteristics, dry matter and yield of corn. Journal of Agriculture, 110, 1-7. (In Persian with English abstract), https://doi.org/10.22092/aj.2016.109214
19. Fowler, B.D., Brydon, J., & Baker, R.J. (1989). Nitrogen fertilization of no till winter wheat and rye. II: Influence of grain protein. Agronomy Journal, 81, 72-77. https://doi.org/2134/agronj1989.00021962008100010013x
20. Garcia-Larez, F.L., Murillo-Hernandez, J.L., Vargas-Sanchez, R.D., Torrescano-Urrutia, G.R., Torres-Martinez, B.D.M., & Sanchez-Escalante, A. (2021). Effect of extraction solvent on metabolites content, antioxidant, and antibacterial activity of coffee bagasse. TIP. Revista especializada en ciencias quimico-biologicas. https://doi.org//10.22201/fssz.23958723e.2021.363
21. Grzesiak, M.T., Grzesiak, S., & Skoczowski, A. (2006). Changes of leaf water potential and gas exchange during and after drought in triticale and maize genotypes differing in drought tolerance. Photosynthetica, 44, 561–568. https://doi.org/1007/s11099-006-0072-z
22. Hajihasani, N., Ameriyan, M.R., Asghari, H.R., & Rahimi, M. (2014). Effect of mycorrhiza, vermicompost and humic acid application on development and yield of Vigna unguiculata M.Sc. Thesis, Shahrood University of Technology, Shahrood, Iran. 135 pp.
23. Haranal, K., Hanumanthappa, M., Mavarkar, N.S., Salimath, S.B., & Nadukeri, S. (2023). Foliar application of humic acid with varied nutrient levels on the performance of sweet corn.‏ International Journal of Research in Agronomy, 6(1), 18-21. https://doi.org/10.33545/2618060x.2023.v6.i1a.157
24. Hashimi, R., Afghani, A.K., Karimi, M.R., & Habibi, H.K. (2019). Effect of organic and inorganic fertilizers levels on spinach (Spinacia oleracea) production and soil properties in Khost Province, Afghanistan. International Journal of Advanced Research, 5(7), 83-87.
25. Hassanen, S.A., Mousa, W.M., & Sultan, F.M. (2020). Effect of foliar application of humic acid, yeast and garlic extracts on growth, yield and quality in forage cowpea. The Middle East Journal, 9(4), 1079-1087. https://doi.org/10.36632/mejar/2020.9.4.84
26. Heidari, M., Paydar, A., Baradarn Firozabad, M., & Abedinin Esfalati, M. (2020). The Effect of drought stress and application of humic on quantitative yield, photosynthetic pigments, and mineral nutrient content in sunflower seeds. Iranian Journal of Field Crop Science, 50(4), 51-62. (In Persian with English abstract). https://doi.org/22059/ijfcs.2018.253008.654448.
27. Hiscox, J.D., & Israelstom, G.F. (1979). A method for the extraction of chlorophyll from leaf tissue without maceration. Canadian Journal of Botany, 57(2), 1332-1339. https://doi.org/10.1139/b79-163.
28. Huculak-Maczka, M., Hoffmann, J., & Hoffmann, K. (2018). Evaluation of the possibilities of using humic acids obtained from lignite in the production of commercial fertilizers. Journal of Soils Sediments, 18, 2868-2880. https://doi.org/10.1007/s11368-017-1907-x  
29. Irigoyen, J.J., Emerich, D.W., & Sanchez-Diaz, M. (1992). Water stress induced changes in concentration of prolun and total soluble sugarsin nodulated alfalfa (Medicago sativa). Plant Physiology, 84, 55-60.
30. Jaleel, C., Manivannan, P., Wahid, A., Farooq, M., Al-Juburi, H., Somasundaram, R., & Panneerselvam, R. (2009). Drought stress in plants: A review on morphological characteristics and pigments composition. International Journal of Agriculture and Biology, 11(2), 43-56.
31. Kahraman, A. (2017). Effect of humic acid doses on yield and quality parameters of cowpea (Vigna unguiculata (L.) Walp) cultivars. Legume Research, 2(1), 1-14. https://doi.org/18805/lr.v0iof.3763 .
32. Kramer, P.S. (1983). Water relation of plants. Academic Press. pp. 342-415.
33. Li, S., Bingshu, H., Wang, J., Liu, J., & Xianmin, H. (2020). Risks of caffeine residues in the environment: Necessity for a targeted Eco pharmacovigilance program. Chemosphere, 243, 125-143. https://doi.org/1016/j.chemosphere.2019.125343 .
34. Lotfi, B., Fotohi, F., Siadat, S.A., & Sadeghi, M. (2018). The effect of nitrogen chemical fertilizer and nitroxin biofertilizer on yield and protein percentage of cowpea seed (Vigna unguiculata). Journal of Crop Ecophysiology, 1(45), 123-138. (In Persian with English abstract).
35. Mahmoodi, R., Nasri, M., & Oveisi, M. (2016). Effects of humic acid spraying on yield and yield components of wheat in drought stress condition. Journal of Crop Ecophysiology, 8(1), 10-26.
36. Maiwan, N., Tunçtürk, M., & Tunçtürk, R. (2023). Effect of humic acid applications on physiological and biochemical properties of soybean (Glycine max) grown under salt stress conditions. Journal of Agricultural Sciences, 33(1), 1-9. https://doi.org/10.29133/yyutbd.1057288
37. Moradi, P., & Amini, K. (2017). Extraction and identification of Urtica dioica l extract and its antibacterial and antifungal properties. Journal of Mazandaran University Medical Science, 27(151), 74-85. (In Persian with English abstract).
38. Nezami, A., Khazaee, H.R., Mirhashemi, S.M., & Hasanzade, F. (2013). The effect of seed hydropriming on germination and seedling growth of maize (Zea mays) in controlled condition. Iranian Journal of Seed Science and Technology, 2(1), 39-54. (In Persian with English abstract).
39. Noghani, F., Kamali, S., & Zaree, G. (2020). Investigation of sensory characteristics and analysis of antioxidant levels of green tea extract and acceptance in minced meat of silver carp (Hypophtalmichthys molitrix) stored at refrigerator temperature 4°C. Advanced Aquaculture Sciences Journal, 3(3), 63-74.
40. Rahmanian, M., & Ayazpour, K. (2020). Evaluation of the effect of salicylic acid, harmal and tea extracts on mechanical transmission of cucumber mosaic virus. Research in Plant Pathology, 6(2), 1-9. (In Persian with English abstract).
41. Rousta, M.J., Enayati, K., Soltani, V., Shiran, M., Ghane, N., Besharat, N., & Neshat, E. (2019). The effects of humic acid application on yield and yield components of wheat and some chemical properties of a saline-sodic soil. Journal of Soil Management and Sustainable Production, 8(4), 95-109. (In Persian with English abstract). http://ejsms.gau.ac.ir.
42. Roy, D., Sarkar, S., Sardar, S., & Sengupta, K. (2017). Effect of nutrient management on the incidence of major insect and disease pests in rice-mustard cropping system. International Journal of Chemical Studies, 5(6), 1408-1412.
43. Sairam, R.K., & Srivastava, G.C. (2001). Water stress tolerance of wheat Triticum aestivum: Variation in hydrogen peroxide accumulation and antioxidant activity in tolerant and susceptible genotypes. Journal of Agronomy and Crop Science, 186, 63-70. https://doi.org/10.1046/j.1439-037x.2001.00461.x
44. Sakinejad, T., Hossaini, S.M., & Hyvari, M. (2011). Calculate changes of bean germination process in the presence of various compounds of biological fertilizer humic acid mixed with micro and macro elements. Journal of American Science, 7(6), 10-14.
45. Shahsavani, S., Abaspour, A., Parsaeian, M., & Unesi, Z. (2017). Effect of fish waste, chemical fertilizer and biofertilizer on yield and yield components of bean (Vigna sinensis) and some soil properties. Iranian Journal of Pulses Research, 8(1), 45-59. (In Persian with English abstract). https://doi.org/22067/ljpr.v8l1.4728
46. Tajik, N., Tajik, M., Mack, I., & Enck, P. (2017). The potential effects of chlorogenic acid, the main phenolic components in coffee, on health: A comprehensive review of the literature. European Journal of Nutrition, 56, 22-35. https://doi.org/1007/s00394-017-1379-1
47. Vigardt, A. (2012). Influence of coffee vermicompost on growth and nutrient quality of greenhouse spinach and field grown green bell peppers. Ph.D. Dissertation, Southern Illinois University Carbondale.

Yoo, Y., Yoo, Y.H., Lee, D.Y., Jung, K.H., Lee, S.W., & Park, J.C. (2023). Caffeine produced in rice plants provides tolerance to water-deficit stress. Antioxidants, 12(11), 19-34.‏ https://doi.org/10.3390/antiox12111984