The effect of different molybdenum levels on some biochemical traits and nitrate accumulation in the spinach plant (Spinacea oleracea L.)

Mafton, Leila; Esna-Ashari, Mahmood; Amerian, Masoomeh

doi:10.22034/iuvs.2023.2013641.1324

The effect of different molybdenum levels on some biochemical traits and nitrate accumulation in the spinach plant (Spinacea oleracea L.)

Document Type : Original Article

Authors

Leila Mafton ¹

Mahmood Esna-Ashari ²

masoomeh amerian ³

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

² Professor, Department of Horticultural Sciences, Faculty of Agriculture, Bu-Ali Sina University, Hamedan, Iran

³ Assistant Professor, Department of Horticultural Sciences and Engineering, Faculty of Agricultural Sciences and Engineering, Razi University, Kermanshah, Iran

10.22034/iuvs.2023.2013641.1324

Abstract

Extended Abstract
1. Introduction: There is significant concern regarding the occurrence of diseases caused by the consumption of vegetables high in nitrate and nitrite, such as spinach. Nitrate accumulation is a common problem in most leafy vegetables and it happens when the amount of nitrate (NO-3) absorption exceeds the amount consumed by the plant. Different nitrogen (N) levels significantly affect the activities and expressions of N metabolism enzymes and concentration of mineral elements in crop plants. Molybdenum (Mo) is an essential micronutrient for plants, playing a critical role in various physiological processes, growth, and development. Although plants require molybdenum in very small amounts compared to macronutrients, it is crucial for several key functions within plants. Molybdenum is an essential component of enzymes involved in nitrogen metabolism and other biochemical processes. Molybdenum deficiency affects the amount of nitrogen in plants, and its presence reduces nitrate accumulation in their tissues. This element is an essential part of the nitrate reductase enzyme necessary to reduce nitrate nitrogen to the amino form, resulting in more and effective utilization of nitrogen by plants. Nitrogenase and nitrate reductase, two enzymes crucial to nitrogen-fixing and nitrogen reduction, need molybdenum to carry out their respective activities. Molybdenum is required as a co-factor for nitrogenase enzymes in symbiotic N fixation, where it catalyzes the redox process that transforms elemental N into ammonium (NH4+) ions. In this study, the effect of different molybdenum concentration on nitrate accumulation in spinach plant under soilless conditions was investigated.
2. Materials and Methods: This study was conducted as a factorial experiment with two factors in a completely randomized design with three replications. The first factor was the concentration of nitrogen in the form of calcium nitrate salt [Ca (NO3) 2.4H2O] at three levels including 0 (control), 10 and 20%. and the second factor was molybdenum concentration in the form of sodium molybdate salt [Na2Mo4O] at four levels including 0 (control), 0.5, 1.5 and 3 μM, all added to the Hoagland's standard solution. Therefore, 12 treatments were applied, and three plants were considered for each replication, so that 108 plants were totally used. Depending on the plants growth rate, the nutrient solution was applied once a day at the beginning of the experiment, but it was increased to 2 times a day at the end of the experiment, when the plants biomass had been increased. The harvesting index in spinach were the size and the number of leaves before flowering, which is recognized as the commercial harvesting stage. Finally, the measured traits included certain biochemical properties, some mineral elements, and the amount of nitrate as well as the activity of nitrate reductase enzyme in root and leaf.
3. Results and Discussion: Based on the obtained results, titratable acidity (TA), soluble solids contents (TSS) and pH increased with increasing molybdenum up to the level of 3 μM, and the simultaneous application of molybdenum and nitrate did not have a significant effect on the mentioned traits. The interaction effect between nitrate and molybdenum on leaf and root nitrate levels, and leaf nitrate reductase enzyme activity was significant at the probability level of 1%. The highest amount of nitrate in leaf (3.2474 g kg-1 FW) and root (2.9191 g k-1g FW) was observed in the combined treatment of 20% nitrate and control molybdenum. On the other hand, the highest level of nitrate reductase enzyme activity was observed when 3 and 1.5 μM molybdenum applied for each of three level nitrate. The highest amount of nitrogen (3.14%), phosphorus (0.064%), iron (0.004%) and potassium (0.78%) elements was obtained from the combined treatment of 3 μM molybdenum and 20% nitrate. The amount of molybdenum element in the control nitrate and 3 μM molybdenum treatment was at its highest level (0.549 mg kg-1 FW).
4. Conclusion: In general, the use of sodium molybdate can be recommended as the way to reduce the accumulation of nitrate and, besides, molybdenum enrichment in spinach increases the nutritional value of this plant. Based on the results, the use of molybdenum in nutrient solution had a reducing effect on nitrate accumulation and an increasing effect on nitrate reductase enzyme activity inside the spinach plant. Therefore, adding 3 μM sodium molybdate to the Hoagland nutrient solution in the hydroponic system, can be recommended for growing spinach to achieve the minimum amount of nitrate accumulation.

Keywords

Nitrate reductase

Nitrogen

Soluble solids

Titratable acidity

Abasifar, A.R, ValizadehKaji, B. & Iravani, M. A. (2020). Effect of green synthesized molybdenum nanoparticles on nitrate accumulation and nitrate reductase activity in spinach. Journal of Plant Nutrition, 43, 13-2. ‏https://doi.org/10.1080/01904167.2019.165934

Abd-El-Latif, F. M., Bakry, K. A., El-Gioushy, S. F., Hussein, A. M. & Mohamed, M. S. (2020). Effect of foliar spray with molybdenum and iron on vagative growth and nutritional status of pear trees. Journal of Plant Production, 11, 655-659.

https://doi.org/10.21608/jpp.2020.110574

Adiloglu, S., Adiloglu, A., Sümer, A. & Satana, A. (2011). Molybdenum Application on the Growth and Nutrient Element Contents of Head Lettuce (Lactuca sativa L.) in Acid Soils. Asian Journal of Chemistry, 23, 937-938.

Ali, M. M. Li, B. Zhi, C. Yousef, A. F. & Chen, F. (2021). Foliar-Supplied Molybdenum Improves Phyto-Nutritional Composition of Leaves and Fruits of Loquat (Eriobotrya japonica Lindl). Agronomy, 11, 1-16.

https://doi.org/10.3390/agronomy11050892

Amerian, M. (2023). Health risk assessment due to heavy metals and nitrates in greenhouse okra of Kermanshah county. Journal of Research in Environmental Health, 8, 406-418. https://doi.org/10.22038/jreh.2023.67299.1539

AOAC. Official method of analysis. Association of official analytical. Chemists, washington, DC. 1990, USA.

Ashoori, S., Abdollahi, F., Jafari, L. (2024). Effect of fertilizer and seed inoculation with Pantoea agglomerans on pigments, vegetative, qualitative, and quantitative characteristics of lettuce (Lactuca sativa). Journal of Vegetables Sciences, 8(1), 50-67. https://doi.org/10.22034/iuvs.2023.2003849.1292

Bambara, S. & Ndakidemi, P. A. (2010). The potential roles of lime and molybdenum on the growth, nitrogen fixation and assimilation of metabolites in nodulated legume: A special reference to Phaseolus vulgaris L. African Journal of Biotechnology, 9, 2482–2489.

‏Biscaro, G. A., Freitas Junior, N. A. D., Soratto, R. P., Kikuti, H., Goulart Junior, S. A. R. & Aguirre, W. M. (2011). Nitrogênio em cobertura e molibdênio via foliar no feijoeiro irrigado cultivado em solo de cerrado. Acta Scientiarum. Agronomy, 33, 665-670. https://doi.org/10.4025/actasciagron.v33i4.6387

Biswas, S., Banerjee, A., Acharyya, P. & Chakraborty, N. (2020). Response of French bean (Phaseolus vulgaris L. cv. Arka Arjun) to Rhizobium inoculation under varied levels of nitrogen and molybdenum. International Journal of Current Microbiology and Applied Sciences, 9, 2759-67.

https://doi.org/10.20546/ijcmas.2020.903.316

Bittner, F. (2014). Molybdenum metabolism in plants and crosstalk to iron. Frontiers in plant science, 5, 1-6. ‏ https://doi.org/3389/fpls.2014.00028

Boussadia, O., Steppe, K., Zgallai, H., Ben El Hadj, S., Braham, M., Lemeur, R. & Van Labeke, M. C. (2010). Effects of nitrogen deficiency on leaf photosynthesis, carbohydrate status and biomass production in two olive cultivars ‘Meski’and ‘Koroneiki’. Scientia Horticulturae, 123, 336–342. https://doi.org/10.1016/j.scienta.2009.09.023

Brengi, S. H. & Abouelsaad, I. A. (2019). The role of different nitrogen sources combined with foliar applications of molybdenum, selenium or sucrose in improving growth and quality of edible parts of spinach (Spinacia oleracea L). Alexandria Science Exchange Journal, 40, 156-168. https://doi.org/10.21608/asejaiqjsae.2019.29731

Cataldo, D. A., Haroon, M., Schrader, L. E. & Youngs, V. L. (1975). Rapid colorimetric determination of nitrate in plant tissue by nitration of salicylic acid. Communications in Soil and Science Plant Analysis, 6, 71–80. https://doi.org/10.1080/00103627509366547

Daniel, R. M. & Curran, M. P. (1981). A method for the determination of nitrate reductase. Journal of Biochemical and Biophysical methods, 4, 131-132. ‏ https://doi.org/10.1016/0165022X(81)90026-9

Elrys, A. S., Abdo, A. I. E. & Desoky, E. S. M. (2018). Potato tubers contamination with nitrate under the influence of nitrogen fertilizers and spray with molybdenum and salicylic acid. Environmental Science and Pollution Research, 25, 7076–89. https://doi.org/10.1007/s11356-017-1075-y

Hassani Moghaddam, E., Bazdar, A. R. & Shaaban, M. (2019). Study of nitrate rate in some vegetables cultivated in Poldokhtar and Khorramabad, Lorestan province. Iranian Journal of Health and Environment, 12, 101-112. (In Persian).

Hille, R. (2013). The molybdenum oxotransferases and related enzymes. Dalton Transactions, 42, 3029–3042.

Jalali, M. & Salehi Chegeni, N. (2020). The Positive Effect of Selenium on Nitrate Accumulation in Spinach (Spinacia oleracea L.) and Lettuce (Lactuca sativa L.). Journal of Horticultural Science, 34, 321-334. https://doi.org/0.22067/jhorts4.v34i2.85540

Kaiser, B. N., Gridley, K. L., Ngaire Brady, J., Phillips, T. & Tyerman, S. D. (2005). The role of molybdenum in agricultural plant production. Annals of botany, 96, 745-754. https://doi.org/10.1093/aob/mci226

Kovács, B., Puskás-Preszner, A., Huzsvai, L., Lévai, L. & Bódi, É. (2015). Effect of molybdenum treatment on molybdenum concentration and nitrate reduction in maize seedlings. Plant Physiology and Biochemistry, 96, 38-44. ‏ https://doi.org/10.1016/j.plaphy.2015.07.013

La Bella, S., Consentino, B. B., Rouphael, Y., Ntatsi, G., De Pasquale, C., Iapichino, G., & Sabatino, L. (2021). Impact of ecklonia maxima seaweed extract and mo foliar treatments on biofortification, spinach yield, quality and nue. Plants (Basel), 10, 1-15. https://doi.org/10.3390/plants10061139

Li, L. I. U., Wei, X. I. A. O., JI, M. L., Chao, Y. A. N. G., Ling, L. I., GAO, D. S. & FU, X. L. (2017). Effects of molybdenum on nutrition, quality, and flavour compounds of strawberry (Fragaria× ananassa Duch. cv. Akihime) fruit. Journal of integrative agriculture, 16, 1502-1512.

Liu, L., Shi, H., Li, S., Sun, M., Zhang, R., Wang, Y. & Ren, F. (2020). Integrated analysis of molybdenum nutrition and nitrate metabolism in strawberry. Frontiers in Plant Science, 11, 1-14. https://doi.org//10.3389/fpls.2020.01117

Longbottom, M. L., Dry, P. R. & Sedgley, M. (2010). Effects of sodium molybdate foliar sprays on molybdenum concentration in the vegetative and reproductive structures and on yield components of Vitis vinifera cv. Merlot. Australian Journal of Grape and Wine Research, 16, 477-490.

Luetic, S., Knezovic, Z., Jurcic, K., Majic, Z., Tripkovic, K. & Sutlovic, D. (2023). Leafy Vegetable Nitrite and Nitrate Content: Potential Health Effects. Foods, 12, 1655.

Machado, R. M. A., Alves-Pereira, I., Lourenço, D. & Ferreira, R. M. A. (2020). Effect of organic compost and inorganic nitrogen fertigation on spinach growth, phytochemical accumulation and antioxidant activity. Heliyon, 6, e05085. ‏ https://doi.org/10.1016/j.heliyon.2020.e05085

Mendel, R. R. (2013). The molybdenum cofactor. Journal of Biological Chemistry, 288, 13165–13172.

https://doi.org/10.1074/jbc.R113.455311

Min, Y. U., Hu, C. X., Sun, X. C. & Wang, Y. H. (2010). Influences of Mo on nitrate reductase, glutamine synthetase and nitrogen accumulation and utilization in Mo-efficient and Mo-inefficient winter wheat cultivars. Agricultural Sciences in China, 9, 355–361.

Mohammadi, H., Kashefi, B. & Khosh ghalb, H. (2017). Investigation of Salicylic acid effect in different growth stages on quality and quantity of Grape varieties rish baba in Shahrood. Journal of Plant Research (Iranian Journal of Biology), 29, 886-895. (In Persian).

https://doi.org/20.1001.1.23832592.1395.29.4.18.6

Moncada, A., Miceli, A., Sabatino, L., Iapichino, G., D’Anna, F. & Vetrano, F. (2018). Effect of molybdenum rate on yield and quality of lettuce, escarole, and curly endive grown in a floating system. Agronomy, 8, 1-16. https://doi.org/10.3390/agronomy8090171

Mondal, S. & Nad, B. K. (2012). Nitrate accumulation in spinach as influenced by sulfur and phosphorus application under increasing nitrogen levels. Journal of plant nutrition, 35, 2081-2088.

Najm, A. A., Hadi, M. R. H. S., Darzi, M. T. & Fazeli, F. (2013). Influence of nitrogen fertilizer and cattle manure on the vegetative growth and tuber production of potato. International Journal of Agriculture and Crop Sciences, 5, 147–154.

Olfati Chirani, J.A., Babalar, M., Kashi, A. A. K., Dadashpour, A. & Shahmoradi, K. (2008). The Effect of ammonium and molybdenum on nitrate concentration in two cultivavs (species) of greenhouse cucumbers. Journal of Horticultural Science (Agricultural Sciences and Technology), 22, 69-77. (In Persian).

Pandey, S. & Dawson, J. (2023). Response of Boron and Molybdenum on Growth and Yield of Black Gram (Vigna mungo L.). International Journal of Plant & Soil Science, 35, 1214-1220.

Rana, M., Bhantana, P., Sun, X. C., Imran, M., Shaaban, M., Moussa, M., Saleem, M. H., Elyamine, A., Binyamin, R., Alam, M. & Afzal, J. (2020). Molybdenum as an essential element for crops: an overview. Biomedical Journal of Scientific & Technical Research, 25, 18535-18547.

Rocha, D. C., da Silva, B. F. I., Moreira dos Santos, J. M., Tavares, D. S., Pauletti, V. & Gomes, M. P. (2020). Do nitrogen sources and molybdenum affect the nutritional quality and nitrate concentrations of hydroponic baby leaf lettuce? Journal of food science, 85, 1605-1612.

Rohilla, P. & Yadav, J. P. (2020). Nitrate reductase structure, role and factors affecting its regulation: a review. Plant Archives, 20, 5787-5793. doi:

Sabatino, L., D'Anna, F., Iapichino, G., Moncada, A., D'Anna, E. & De Pasquale, C. (2019). Interactive effects of genotype and molybdenum supply on yield and overall fruit quality of tomato. Frontiers in Plant Science, 9, 1-10.

https://doi.org/10.3389/fpls.2018.01922

Seilsepour, M. (2020). Study of nitrate concentration in Varamin plain leafy vegetables and evaluation of its risk for human. Horticultural Plants Nutrition, 3, 69- 86.

Seilsepour, M. (2022). Study of the Effect of Municipal Solid Waste Compost and Nitrogen on Spinach Yield, Soil and Spinach Nitrate Content and Evaluation of Its Risk Aversion for Human Health, Journal of Vegetables Sciences, 6(1),. 158-175.

Soares Filho, S. I. B., Lazarini, E., Júnior, V. O. & Bernardes, J. V. S. (2020). Sowing dates and molybdenum foliar application for two peanut cultivars. Revista Ciência Agrícola, 18, 27-34.

Soltani, A., Robertson, M. J. & Manschadi, A. M. (2006). Modeling chickpea growth and development: Nitrogen accumulation and use. Field Crops Research, 99, 24-34.

https://doi.org/10.1016/j.fcr.2006.02.006

Song, W., Yue, L., Fan, X., Luo, Y., Ying, B., Sun, S., Zheng, D., Liu, Q., Hamdy, M. S. & Sun, X. (2023). Recent progress and strategies on design of catalysts for electrochemical ammonia synthesis from nitrate reduction. Inorganic Chemistry Frontiers, 10, 3489-3514.

https://doi.org/10.1039/D3QI00554B

Steiner, F., Zoz, T., Zuffo, A. M., Pereira-Machado, P., Zoz, J. & Zoz, A. (2018). Foliar application of molybdenum enhanced quality and yield of crispleaf lettuce (Lactuca sativa L., cv. Grand Rapids). Acta Agronómica, 67, 73-78.

https://doi.org/10.15446/acag.v67n1.59272

Swami, Y. P., Waikar, S. L. & Sallawar, S. C. (2021). Impact of ammonium molybdate supplementation on growth, yield and nutrient uptake of pigeon pea (Cajanus cajan). Journal of Pharmacognosy and Phytochemistry, 10, 2145-2150. ‏

Sym, G. J. (1984). Optimisation of the in‐vivo assay conditions for nitrate reductase in barley (Hordeum vulgare L. cv. Igri). Journal of the Science of Food and Agriculture, 35, 725-730.

https://doi.org/10.1002/jsfa.2740350703‏

Wang, M., Hasegawa, T., Beier, M., Hayashi, M., Ohmori, Y., Yano, K., Teramoto, S., Kamiya, T. & Fujiwara, T. (2021). Growth and nitrate reductase activity are impaired in rice osnlp4 mutants supplied with nitrate. Plant and Cell Physiology, 62, 1156-1167. doi: 10.1093/pcp/pcab035

Wang, Y., Gong, Z., Friml, J. & Zhang, J. (2019). Nitrate Modulates the Differentiation of Root Distal Stem Cells. Plant Physiology, 180, 22-25.

https://doi.org/10.1104/pp.18.01305

Zeka, N., Mero, G., Skenderasi, B. & Gjançi, S. (2014). Effects of nitrogen sources and levels on yield and nutritive values of spinach (Spinacia oleracea L). Journal of International Academic Research for Multidisciplinary, 2, 327–337.