Evaluation and Determination of Lettuce (Lactuca sativa L.) Nitrogen Status Using Chlorophyll Meter under Greenhouse Conditions

Document Type : Research Paper


1 Horticultural Sciences Department, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran.

2 Department of Irrigation & Reclamation Engineering Faculty of Agriculture Engineering &Technology College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran.


Leaves Chlorophyll content can be used as an indicator to determine nitrogen status in plants. The soil–plant analyses development (SPAD) meter is one of the most widely used tools in crop nitrogen management studies. The current research was aimed to investigate crop nitrogen status, introduce a standardized index called the normalized chlorophyll meter index (NSPAD) and find its relationship with nitrogen nutrition in lettuce. For this purpose, lettuce was cultivated in the research greenhouse of the College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran during two periods. Different nitrogen fertilizer treatments were applied at six levels with three replications. The values of dry matter, nitrogen concentration and chlorophyll were measured seven times during the growth period. Nitrogen nutrition indices (NNI) and NSPAD were calculated for the first period and the NSPAD-NNI relationship was extracted. Then the obtained relationship was verified by the data of the second cultivation. The results showed that the chlorophyll meter readings and nitrogen concentration have a positive correlation with the increase in the level of fertilizer application. Also, the validation results of NSPAD-NNI relationship showed a good performance with R2, MBE and RRMSE values equal to 0.79, -0.01 and 11.41%, respectively. Therefore, the developed relationship can be used as a fast, non-destructive and reliable approach for sustainable nitrogen fertilizer management of lettuce grown under greenhouse conditions.


Adhikari, C., Bronson, K., Panuallah, G., Regmi, A., Saha, P., Dobermann, A., et al. (1999). On-farm soil N supply and N nutrition in the rice–wheat system of Nepal and Bangladesh. Field Crops Res. 64, 273–286.doi: 10.1016/S03784290(99)00063-5
Ata-Ul-Karim, S., Cao, Q., Zhu, Y., Tang, L., Rehmani, M. and Cao, W., (2016). Non-destructive Assessment of Plant Nitrogen Parameters Using Leaf Chlorophyll Measurements in Rice. Frontiers in Plant Science, 7.
Ata-Ul-Karim, S. T., Liu, X., Lu, Z., Zheng, H., Cao, W., and Zhu, Y. (2017). Estimation of nitrogen fertilizer requirement for rice crop using critical nitrogen dilution curve. Field Crops Res. 201, 32–40.doi: 10.1371/journal.pone. 0104540
Barutcular, C., Yıldırım, M., Koç, M., Akıncı, C., Toptaş, I., Albayrak, O., et al. (2016). Evaluation of SPAD chlorophyll in spring wheat genotypes under different environments. Fresenius Environmental Bulletin, 25(4), 1258–1266.
Blackmer, T.M., Schepers, J.S., Varvel, G.E., (1994). Light reflectance compared nitrogen stress measurements in corn leaves. Agron. J. 86,934–938.
Conversa G, Elia A. (2019) Growth, Critical N Concentration and Crop N Demand in Butterhead and Crisphead Lettuce Grown under Mediterranean Conditions. Agronomy. 9(11):681.
Debaeke, P., Rouet, P., and Justes, E. (2006). Relationship between the normalized SPAD index and the nitrogen nutrition index: application to durum wheat. J. Plant Nutr. 29, 75–92.doi: 10.1080/01904160500416471
Djidonou, D., & Leskovar, D. (2019). Seasonal Changes in Growth, Nitrogen Nutrition, and Yield of Hydroponic Lettuce. HortScience. 54. 76-85. 10.21273/HORTSCI13567-18.
FAOSTAT, 2021. Fao.org. URL http://www.fao.org/faostat.
Fox, R.H., Piekielek, W.P., Macneal, K.M., (1994). Using a chlorophyll meter to predict nitrogen fertilizer needs of winter wheat. Commun. Soil Sci. Plant Anal. 25, 171–181.
Habibi R., Delshad M., Rahimikhoob H. (2022). Determination of critical nitrogen concentration curve based on dry matter and leaf area index of Lettuce (Lactuca sativa L.) grown under greenhouse conditions. Master of Science Thesis.
Huang, J., F. He, K. Cui, R.J. Buresh, B. Xu, W. Gong, and S. Peng. (2008). Determination of optimal nitrogen rate for rice varieties using a chlorophyll meter. Field Crops Res. 105:70–80. doi:10.1016/j. fcr.20 07.07.0 06
Jamieson PD, Porter JR, Wilson DR (1991) A test of computer simulation model ARC-WHEAT1 on wheat crops grown in New Zealand. Field Crops Res. 27: 337–350.
Justes, E., Mary, B., Meynard, J. M., Machet, J. M., and Thelier-Huche, L. (1994). Determination of a critical nitrogen dilution curve for winter wheat crops. Ann. Bot. 74, 397–407.doi: 10.1006/anbo.1994.1133
Lemaire, G., Meynard, J.-M., (1997). Use of the nitrogen nutrition index for the analysis of agronomical data. In: Lemaire, G. (Ed.), Diagnosis of the Nitrogen Status in Crops, pp. 45–55.
Lemaire, Gilles, Marie-Hélène Jeuffroy, and François Gastal. (2008). Diagnosis Tool for Plant and Crop N Status in Vegetative Stage: Theory and Practices for Crop N Management. European Journal of Agronomy .28 (4): 614–24.
Madakadze, I. C., K. A. Stewart, R. M., Madakadze, P. R. Peterson, B. E. Coulman, D. L. Smith. 1999.  Field evaluation of the chlorophyll meter to predict yield and nitrogen concentration of             switchgrass. J. Plant Nutr. 22 (6): 1001-1010.
Muniandy JM, Yusop Z, Askari M (2016) Evaluation of reference evapotranspiration models and determination of crop coefficient for Momordica charantia and Capsicum annuum. Agric. Water Manage. 169:77–89
Pavlou GC, Ehaliotis CD, Kavvadias VA. (2007) Effect of organic and inorganic fertilizers applied during successive crop seasons on growth and nitrate accumulation in lettuce. Sci Hortic. 111: 319–325.
 Prohens, J. and Nuez, F., 2008. Vegetables. New York: Springer Chang, S. and Robison, D., 2003. Nondestructive and rapid estimation of hardwood foliar nitrogen status using the S 50 chlorophyll meter. Forest Ecology and Management, 181(3), pp.331-338.
Rahimikhoob, H., Sohrabi, T. and Delshad, M. (2020a) Development of a Critical Nitrogen Dilution Curve for Basil (Ocimum basilicum L.) Under Greenhouse Conditions. Journal of Soil Science and Plant Nutrition. https://doi.org/10.1007/s42729-020-00174-5
Rahimikhoob, H., Sohrabi, T. and Delshad, M. (2020b) Estimation of In-season Basil Nitrogen Requirement Using the Nitrogen Nutrition Index. Iranian journal of soil and water research. 51(8)-2039-2049.
Ravier, C., Quemada, M. and Jeuffroy, M., (2017). Use of a chlorophyll meter to assess nitrogen nutrition index during the growth cycle in winter wheat. Field Crops Research, 214, pp.73-82.
Santhi C, Arnold JG, Williams JR, Dugas WA, Srinivasan R, Hauck LM (2001) Validation of SWAT model on a large river basin with point and nonpoint source. Wiley online libaray.
Still, D.W. (2007). Lettuce. In: Kole, C. (eds) Vegetables. Genome Mapping and Molecular Breeding in Plants, vol 5. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-34536-7_2
Umar S, Anjana Anjum NA, Khan NA (2013) Nitrate management approaches in leafy vegetables. In: Umar S, Anjum NA, Khan NA (eds) Nitrate in leafy vegetables: toxicity and safety measures. IK International Publishing House Pvt. Ltd, New Delhi, pp 166–181
Yan X., Gong W (2010). The role of chemical and organic fertilizers on yield, yield variability and carbon sequestration— results of a 19year experiment. Plant Soil 331(1–2):471–480. https://doi.org/10. 1007/s11104-009-0268-7
Yuan, Z., Ata-Ul-Karim, S. T., Cao, Q., Lu, Z., Cao, W., Zhu, Y., et al. (2016). Indicators for diagnosing nitrogen status of rice based on chlorophyll meter readings. Field Crops Res. 185, 12–20.doi: 10.1016/j.fcr.2015.10.003
Yue, X., Hu, Y., Zhang, H. et al. Evaluation of Both SPAD Reading and SPAD Index on Estimating the Plant Nitrogen Status of Winter Wheat. Int. J. Plant Prod. 14, 67–75 (2020). https://doi.org/10.1007/s42106-019-00068-2
Zhao, B. (2014). Determining of a critical dilution curve concentration for plant nitrogen in winter barley. Field Crops Res. 160, 64–72.
Zhao, B., Liu, Z., Ata-Ul-Karim, S. T., Xiao, J., Liu, Z., Qi, A., et al. (2016). Rapid and nondestructive estimation of the nitrogen nutrition index in winter barley using chlorophyll measurements. Field Crops Res. 185, 59–68.doi: 10.1016/j.fcr.2015.10.021
Zhao, B., Ata-Ul-Karim, S., Liu, Z., Zhang, J., Xiao, J., Liu, Z., Qin, A., Ning, D., Yang, Q., Zhang, Y. and Duan, A., 2018. Simple Assessment of Nitrogen Nutrition Index in Summer Maize by Using Chlorophyll Meter Readings. Frontiers in Plant Science, 9.