Optimization of nitrogen fertilizer and plant spacing on the row parameters in ‎spinach cv. “Harrier” using response surface methodology

Document Type : Full Paper


1 Ph. D. Candidate, University College of Agriculture & Natural Resources, University of ‎Tehran, Karaj, Iran

2 Associate Professor. University College of Agriculture & Natural Resources, University of Tehran, Karaj, ‎Iran

3 Assistant Professor, Sugar Beet Research Department, Hamedan Agricultural and Natural Resources Research and Education ‎Center, AREEO, Hamedan, Iran

4 Assistant Professor, University College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran


In this study, optimum level of nitrogen fertilizer and plant spacing on the row were investigated in spinach cv.”Harrier” based on the aims of four scenarios: economic, health of food, marketable and optimum production, using statistical response surface methodology. The independent variables including plant spacing on the row (7, 11 and 15 cm) and nitrogen fertilizer (0, 200 and 400 kg urea ha-1), were evaluated using central composite design at Research Field of University of Tehran. Leaf fresh weight, biomass, leaf area index, nitrogen use efficiency, nitrate concentration and total chlorophyll traits were considered as dependent variables. The fitting of data to second order polynomial equation and optimization of responses was performed using Mnitab16 software. Increasing of plant spacing on the row had alone a significant effect on total chlorophyll traits. High levels of nitrogen fertilizer had an increasing effect on dependent variables, except nitrogen use efficiency. The optimum combination of variables and desirability function for economic, environmental, marketable and optimal production limit scenarios were at plant spacing on the row of 7, 7.48, 7.24 and 7 cm, nitrogen fertilizer of 189.9, 60.6, 189.9 and 149.5 kg ha-1, and 0.97, 0.77, 0.9 and 0.82, respectively. Based on desirability function results, the model possesses a high power to simulate optimum level of independent variables at economic scenario than the others.


  1. Abrol, Y. P., Chatterjee, S. R., Kumar, P. A. & Jain, V. (1999). Improvement in nitrogen use efficiency: physiological and molecular approaches. Current Science, 76, 1357-1364.
  2. Ahmadil, H., Akbarpour, V., Dashti, F. & Shojaeian, A. (2010). Effect of different levels of nitrogen fertilizer on yield, nitrate accumulation and several quantitative attributes of five Iranian spinach American-Eurasian journal of Agriculture and Environmental Science, 8(4), 468-473.
  3. Andrade, F. H., Calvino, P., Cirilo, A. & Barbieri, P. (2002). Yield responses to narrow rows depend on increased radiation interception. Agronomy Journal, 94: 975-980.
  4. Arnon, D. I. (1949). Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiology, 24, 1-15.
  5. Aslan, N. (2007). Application of response surface methodology and central composite rotatable design for modeling the influence of some operating variables of a Multi-gravity Separator for coal cleaning. Fuel, 86, 769-776.
  6. Bansal, G. L., Rana, M. C. & Upadhyay, R. G. (1995). Response of grain amaranth (Amaranthus hypochondriacus) to plant density. Indian Journal of Agricultural Sciences, 65(11), 818-820.
  7. Biemond, H. (1995). Effects of nitrogen on development and growth of the leaves of vegetables. 3. Appearance and expansion growth of leaves of spinach. Netherlands Journal of Agricultural Science, 43, 247-260.
  8. Biemond, H., Vos, J. & Struik, P. C. (1996). Effects of nitrogen on accumulation and partitioning of dry matter and nitrogen of vegetables. 3. Spinach. Journal of Agricultural Science, 44, 227-239.
  9. Box, G. E. P. & Hunter, J. S. (1957). Multi-factor experimental design for exploring response surfaces. Annals of Mathematical Statistics, 28, 195-241.
  10. Bracey, R.P., Parish, R. L., Bergeron, P. E. & Constantin, R. J. )1990(. High density planting for maximizing yields in greens. Louisiana Agriculture, 34(2), 6-7.
  11. Cantliffe, D. J. (1973). Nitrate accumulation in table beets and spinach as affected by nitrogen, phosphorus, and potassium nutrition and light intensity. Agronomy Journal, 65, 563-565.
  12. Cataldo, D. A., Haroon, M., Schrader, L. E. & Youngs, V. L. (1975). Rapid colorimetric of nitrate in plant tissue by nitration of salicylic. Soil Science and Plant Analysis, 6 (1), 71-80.
  13. Chardon, F., Barthélémy, J., Daniel-Vedele, F. & Masclaux-Daubresse, C. (2010). Natural variation of nitrate uptake and nitrogen use efficiency in Arabidopsis thaliana cultivated with limiting and ample nitrogen supply. Journal of Experimental Botany, 61(9), 2293-2302.
  14. Dimri, D. C. & Gulshan, L. A. L. (1997). Effect of nitrogen fertilization spacing and method of planting on yield parameters and quality of tomato cultivar ‘Pant Banar’. Vegetable Science, 15(2), 105-112.
  15. Dong, H., Li, W., Tang, W., Li, Z., Zhang, D. & Niu, Y. (2006). Yield, quality and leaf senescence of cotton grown at varying planting dates and plant densities in the Yellow River Valley of China. Field Crops Research, 98, 106–115.
  16. Dreccer, M. F. (2006). Nitrogen use at the leaf and canopy level: A framework to improve crop N use efficiency. Journal of Crop Improvement, 15(2), 97-125.
  17. Evans, J. R. & Terashima, I. (1987). Effects of nitrogen nutrition on electron transport components and photosynthesis in spinach. Functional Plant Biology, 14, 59-68.
  18. Gutrierrez-Rodriguez, E., Lieth, H. J., Jernstedt, J. A., Labavitch, J. M., Suslow, T. V. & Cantwell, M. I. (2013). Texture, composition and anatomy of spinach leaves in relation to nitrogen fertilization. Journal of the Science of Food and Agriculture, 93, 227-237.
  19. Goel, J., Kadirvelu, K., Rajagopal, C. & Garg V. K. (2006). Cadmium (II) uptake from aqueous solution by adsorption onto carbon aerogel using a response surface methodological approach. Industrial and Engineering Chemistry Research, 45(19), 6531-6537.
  20. Hamidi A. & A. Dabagh Mohamadinasab. (1996). Evaluation of grain yield and its components, biomass and harvest index in two corn hybrids in plant density and nitrogen rates. Agricultural Science Journal, 10, 39-53.
  21. Heuvelink, E. (2005). Tomatoes. (pp. 339). CABI Publishing.
  22. Heins, B. & Schenk, M. (1987). Nitrogen. Journal of Plant Nutrition, 10(9),1743-1751.
  23. Hirel, B., Le Gouis, J., Ney, B. & Gallais, A. (2007). The challenge of improving nitrogen use efficiency in crop plants: towards a more central role for genetic variability and quantitative genetics within integrated approaches. Journal of Experimental Botany, 58(9), 2369-2387.
  24. Jamieson, P. D., Porter, J. R., & Wilson, D. R. (1991). A test of the computer simulation model ARCWHEAT1 on wheat crops grown in New Zealand. Field Crops Research, 27, 337–
  25. Kant, S., Bi, Y.M. & Rothstein, S. (2011). Understanding plant response to nitrogen limitation for the improvement of crop nitrogen use efficiency. Journal of Experimental Botany, 62(4), 1499-1509.
  26. Kalavathy, H. M., Regupathib, I., Pillai, M. G. & Miranda, L. R. (2009). Modelling, analysis and optimization of adsorption parameters for H3PO4 activated rubber wood sawdust using response surface methodology (RSM). Colloids and Surfaces B: Biointerfaces, 70, 35–
  27. Koocheki, A., Nassiri Mahallati, M., Moradi, R., Mansouri, H. (2012). Optimization of water, nitrogen and density in canola cultivation by central composite design. Journal of Agroecology, 1(3),1 -16. (in Farsi).
  28. Law-Ogbomo, K. E. & Egharevba, R. K. A. (2009). Effects of planting density and NPK fertilizer application on yield and yield components of tomato (Lycospersicon esculentum Mill) in forest Location. World Journal of Agricultural Sciences, 5(2), 152-158.
  29. Lefsrud, M. G., Kopsell, D. A. & Kopsell, D. E. (2007). Nitrogen levels influence biomass, elemental accumulations, and pigment concentrations in spinach. Journal of Plant Nutrition, 30(2), 171-185.
  30. Li, P., Dong, H., Zheng, C., Sun, M., Liu, A., Wang, G., Liu, S., Zhang, S., Chen, J., Li, Y., Pang, C. & Zhao, X. (2017). Optimizing nitrogen application rate and plant density for improving cotton yield and nitrogen use efficiency in the North China Plain. Plos One, 12(10), 1-15.
  31. Liu, C., Sung, Y., Chen, B. & Lai, H. (2014). Effects of nitrogen fertilizers on the growth and nitrate content of lettuce (Lactuca sativa). International Journal of Environmental Research and Public Health, 11, 4427-4440.
  32. Machado, R. M. A., Alves-Pereira, I. & Ferreira, R. M. A. (2018). Plant growth, phytochemical accumulation and antioxidant activity of substrate-grown spinach. Heliyon, 4, e00751.
  33. Majeed, A., Muhmood, A., Niaz, A., Javid, S, Ashfaq Ahmad, Z., Hussain Shah, S. S., Hussain Shah, A. (2015). Bed planting of wheat (Triticum aestivum) improves nitrogen use efficiency and grain yield compared to flat planting. The Crop Journal, 3, 118-124.
  34. Mansouri, H., Banayan Aval, M., Rezvani Moghaddam, P. & Lakzian, A. (2014a). Management of nitrogen, irrigation and planting density in persian shallot (Allium hirtifolium) by using central composite optimizing method. Journal of Agricultural Science and Sustainable Production, 41-60. (in Farsi).
  35. Mansouri, H., Banayan, M., Rezvani Moghaddam, P. & Lakzian, A. (2014b). Management of nitrogen fertilizer, irrigation and plant density in onion production using response surface methodology as optimization approach. African Journal of Agricultural Research, 9(7), 676-687.
  36. Mansouri, H., Rael, Y., Nokhbe Zaeim, A. (2015). Simulating future wheat yield under climate change, carbon dioxide enrichment and technology improvement in Iran. Case study: Azarbaijan region. Spanish Journal of Agricultural Research, 13(4),1-12.
  37. Moll, R. H., Kamprath, E. J. & Jackson, W. A. (1982). Analysis and interpretation of factors which contribute to efficiency of nitrogen utilization. Agronomy Journal, 74, 562-564.
  38. Mondal, S. & Nad, B. K. (2012). Nitrate accumulation in spinach as influenced by sulphur and phosphorus application under increasing nitrogen levels. Journal of Plant Nutrition, 35 (14), 2081-2088.
  39. Nash, J. E. & Sutcliffe, J. V. (1970). River flow forecasting through conceptual models. Part I: A discussion of principles. Journal Hydrology, 10, 282-290.
  40. Neeteson, J. J. & Carton, O. T. (1999). The environmental impact of nitrogen in field vegetable production. Acta Horticulturae, 563, 21-28.
  41. Okazaki, K., Oka, N., Shinano, T., Osaki, M. & Takebe, M. (2008). Differences in the metabolite profiles of spinach (Spinacia oleracea) leaf in different concentrations of nitrate in the culture solution. Plant Cell Physiology, 49(2), 170-177.
  42. Peoples, M. B., Freney, J. R. & Mosier, A. R. (1995). Grapes. In: P.E. Bacon (Ed), Nitrogen fertilization in the environment. (pp. 565-602.) New York: Marcel Dekker Inc.
  43. Santamaria, P. (2006). Nitrate in vegetables: toxicity, content, intake and EC regulation. Journal of the Science of Food and Agriculture, 86, 10-17.
  44. Sharma, S. K. (1994). Effect of nitrogen and spacing on plant growth seed yield and quality of spinach seed. Annals of Agricultural Research, 15, 462-464.
  45. Sinclair, T. R. & Muchow, R. C. (1995). Effect of nitrogen supply on maize yield: I. modeling physiological responses. Agronomy Journal, 87(4), 632-641.
  46. Stagnari, F., Bitetto, V. D. & Pisante, M. (2007). Effects of N fertilizers and rates on yield, safety and nutrients in processing spinach genotypes. Scientia Horticulturae, 114, 225-233.
  47. Sun, Y. J., Sun, Y. Y., Li, X. Y., Guo, X. & Ma, J. (2009). Relationship of nitrogen utilization and activities of key enzymes involved in nitrogen metabolism in rice under water-nitrogen interaction. Acta Agronomica Sinica, 35, 2055-2063.
  48. Tafteh, A. & Sepaskhah, A. )2012(. Yield and nitrogen leaching in maize field under different nitrogen rates and partial root drying irrigation. International Journal of Plant Production, 6, 93-113.
  49. Wang, Z. & Li, S. (2007). Effects of nitrogen and phosphorus fertilization on plant growth and nitrate accumulation in vegetables. Journal of Plant Nutrition, 27(3), 539-556.
  50. Wolfe, A. & Patz, J. A. (2002). Reactive nitrogen and human health: acute and long-term implications. Journal of the Human Environment, 31(2), 120-125.
  51. Xu, G., Fan, X. & Miller, A. J. (2012). Plant nitrogen assimilation and use efficiency. Annual Review of Plant Biology, 63, 153-182.
  52. Zhang, J., Sha, Z., Zhang, Y., Bei, Z. & Cao, L. (2015). The effects of different water and nitrogen levels on yield, water and nitrogen utilization efficiencies of spinach (Spinacia oleracea). Canadian Journal of Plant Science, 95(4), 671-679.