Document Type : Research Paper


1 M.Sc. Student, Plant Breeding and Biotechnology Department, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.

2 Associate Professor, Plant Breeding and Biotechnology Department, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran

3 Associate Professor, Biotechnology Research Center, Shiraz University, Shiraz, Iran

4 Assistant Professor, Rice Research Institute of Iran (RRII), Agricultural Research Education and Extension Organization (AREEO) Rasht, Rasht, Iran.

5 Research Assistant Professor, Seed and Plant Improvement Research Department, Bushehr Agricultural and Natural Resources Research and Education Center, AREEO, Bushehr, Iran.


In this study, in order to investigate the response to salinity stress, seedlings of two rice kinds, Hashemi and its advanced mutant line, have exposed to 100 mM NaCl as a salinity stress in an experiment, conducted in 2017 at Gorgan University of Agricultural Sciences and Natural Resources, Iran. For the biochemical investigation, root sampling is performed during three and six days after the salinity stress treatment. The experiment is conducted as a split plot with randomized complete block design with three replications in hydroponic culture. Under salinity stress, the sodium ion content in both genotypes’ roots has increased significantly, while this trend is much lower in the root of mutant genotype than wild type. The induced oxidative stress of salinity stress is measured by the amount of hydrogen peroxide, indicating that the wild type is under higher oxidative stress which is confirmed by the higher amount of malondialdehyde. Evaluation of antioxidant enzymes’ activity include superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase. It reveals a significant rise in the root of the mutant genotype. Overall, this study shows that mutation in the rice genotype leads to salt tolerance, compared to the wild type, through promoting the activity of oxidative enzymes and the synthesis of some osmolytes in the root tissue.


Aebi, H., & Lester, P. (1984). Catalase in vitro. Methods in enzymology. 121–126. https,//
Ashraf, M., & Foolad, M. R. (2007). Roles of glycinebetaine and proline in improving plant abiotic stress tolerance. Environmental and Experimental Botany. 59, 206–216. https,//
Bao, J. Y., Lee, S., Chen, C., Zhang, X. Q., Zhang, Y., Liu, S. Q., Clark, T., Wang, J., Cao, M. L & Yang, H. M (2005). Serial analysis of gene expression study of a hybrid rice strain (LYP9) and its parental cultivars. Plant Physiology.138, 1216-1231. 10.1104/pp.105.060988   
Bates, L. S., Waldren, B. P & Teare, I. D. (1973). Rapid determination of free proline of water-stress studies. Plant and Soil. 39,205–207. http,//
Beyer, W. F, & Fridovich, I (1987). Assaying for superoxide dismutase activity, some large consequences of minor changes in conditions. Analytical Biochemistry. 161, 559-566. https,//
Broadbent, P., Creissen, G. P., Kular, B., Wellburn, A. R. & Mullineaux, P. M. (1995). Oxidative stress responses in transgenic tobacco containing altered levels of glutathione reductase activity. The plant journal. 8, 247-255. 10.1046/j.1365-313X.1995. 08020247.x
Chutipaijit, S., Cha-Um, S., & Sompornpailin, K. (2009). Differential accumulation of proline and flavonoids in indica rice varieties against salinity. Pakistan Journal of Botany. 41, 2497-2506.
Ebadi, A. A. (2018) Evaluation of rice mutant lines under drought stress. Final report of the research method. Registration number 53323. Agricultural Research, Training and Extension Center.
Flowers, T. J. (2004) Improving crop salt tolerance. Journal of Experimental Botany, 55, 307-319.
Forlani, G., Bertazzini, M., and Cagnano, G. (2018). Stress-driven increase in proline levels, and not proline levels themselves, correlates with the ability to withstand excess salt in a group of 17 Italian rice genotypes. Plant Biology. 2, 336-34210.1111/plb.12916.
Forough, M., Navabpour, S., Ebrahimie, E., Ebadi, A., Kiani, D. (2018). Evaluation of salinity response through the antioxidant defense system and osmolyte accumulation in a mutant rice. Journal of Plant Molecular Breeding. 6(2), 27-37. doi, 10.22058/jpmb.2019.114746.1192
Gill, S. S. & Tuteja, N. (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology Biochemistry. 48,909–930. 10.1016/j.plaphy.2010.08.016.
Grattan, S. R., Zeng, L., Shannon, M. C & Roberts, S. R. (2002). Rice is more sensitive to salinity than previously thought. 2002. California Agriculture. 56, 189–195. https,// v056n06p189
Habibollahi, N., Mahdiyeh, M & Amirjani, M.R. (2012). Effect of salt stress on growth, proline, antioxidant enzyme activity and photosystem II efficiency in salt-sensitive and -tolerant rice cultivars. Journal of Plant Biology. 13, 1-7 (Persian).
Hazman, M., Hause, B., Eiche, E., Nick, P., & Riemann, M. (2015). Increased tolerance to salt stress in OPDA-deficient rice ALLENE OXIDE CYCLASE mutants is linked to an increased ROS-scavenging activity. Journal of Experimental Botany. 66,3339–3352. 10.1093/jxb/erv142
Heath, R. L., & Packer, L. (1968). Photoperoxidation in isolated chloroplasts, I. Kinetics and stoichiometry of fatty acid peroxidation. Archives of Biochemistry and Biophysics, 125,189-198. https,//
Jayakannan, M., Bose, J., Babourina, O., Rengel, Z., & Shabala, S. (2015). Salicylic acid in plant salinity stress signaling and tolerance. Plant growth regulation. 76,25-40.
Kaya, C., Akram, N., Ashraf, M., & Sonmez, O. (2018). Exogenous application of humic acid mitigates salinitystress in maize (Zea mays L.) plants by improving some key physico-biochemical attributes. Cereal Research Communications. 46, 67-78. 10.1556/0806.45.2017.064
Khan, M. H. & Panda, S. K. (2008). Alterations in root lipid peroxidation and antioxidative responses in two rice cultivars under NaCl-salinity stress. Acta Physiologiae Plantarum. 30,81–89. 10.1007/s11738-007-0093-7
Kiani, D., Soltanloo, H., Ramezanpour, S. S., Nasrolahnezhad Qumi, A. A., Yamchi, A., Zaynali Nezhad, Kh., & Tavakol, E. (2017). A barley mutant with improved salt tolerance through ion homeostasis and ROS scavenging under salt stress. Acta Physiologiae Plantarum, 39, 90. 10.1007/s11738-017-2359-z
Kibria, M. G., Hossain, M., Murata, Y.,  & Anamul Hoque, M. D. (2017). Antioxidant defense mechanisms of salinity tolerance in rice genotypes. Rice Science. 24, 155-162. https,//
Koca, H., Bor, M., Özdemir, F., & Türkan, I. (2007). The effect of salt stress on lipid peroxidation, antioxidative enzymes and proline content of sesame cultivars. Environmental and Experimental Botany. 60,344–51 https,//
Kordrostami, M., Rabiei, B. & Kumleh, H.H. (2017). Different physiobiochemical and transcriptomic reactions of rice (Oryza sativa L.) cultivars differing in terms of salt sensitivity under salinity stress. Environmental Science and Pollution Research. 24, 7184. https,//
Lin, K.C., Jwo, W.S., Chandrika, N., Wu, T.M., Lai, M.H., Wang, C.S., & Hong C.Y. (2016). A rice mutant defective in antioxidant-defense system and sodium homeostasis possess increased sensitivity to salt stress. Biologia Plantarum. 60, 86-94.
Meloni, D. A., Oli, M. A., & Martinez, C. A. (2003). Photosynthesis and activity of superoxide dismutase, peroxidase and glutathione reductase in cotton under salt stress. Environmental and Experimental Botany. 49,69–76. https,//
Nabiollahi, K., Taghizadeh-Mehrjardi, R., Kerry, R. & Moradian, S. (2017). Assessment of soil quality indices for salt-affected agricultural land in Kurdistan Province, Iran. Ecological Indicators. 83, 482-494.
Nakano, Y., & Asada, K. (1981). Hydrogen peroxide is scavenged by ascorbat specific peroxidase in spinach chloroplast. Plant and cell physiology. 22, 867-880. https,//
Saeedipour, S. (2013). Relationship of grain yield, ABA and proline accumulation in tolerant and sensitive wheat cultivars as affected by water stress. Proceedings of the National Academy of Sciences of the United States of America. 83, 311–315. 10.1007/s40011-012-0147-5
Sagisaka, S. (1976). The occurrence of peroxide in a perennial plant Populas gelrica. Plant Physiology. 57,308-309.
Schnell, D. M. & Clair, D. St. (2014). Redox pioneer, Professor Joe M. McCord. Antioxidants and Redox Signaling. 20, 183–188. https,//
Shavrukov, Y. (2012). Salt stress or salt shock, which genes are we studying? Journal of Experimental Botany. 64,119-127. 10.1093/jxb/ers316
Shu, Q. Y., & Lagoda, P. J. L. (Ed.). (2007). Mutation techniques for gene discovery and crop improvement, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture.
Singh, R., & Flowers, T. (2010). Physiology and molecular biology of the effects of salinity on rice. In, M. Pessarakli (eds.) Handbook of Plant and Crop Stress. 899–939 pp., CRC Press, Boca Raton, USA.
Smith, I.K., Vierheller, T. L. & Thorne, C. A. (1988). Assay of glutathione reductase in crude tissue homogenates using 5,5-dithiobis (2-nitrobenzoic acid). Analytical Biochemistry.175, 408-413. 10.1016/0003-2697(88)90564-7
Sreenivasulu, N., Grimm, B., Wobus, U., & Weschke, W. (2000). Differential response of antioxidant compounds to salinity stress in salt tolerant and salt-sensitive seedlings of foxtail millet (Setaria italica). Physiology Plant. 109,435–442. 10.1034/j.1399-3054.2000. 100410.x
Vaidyanathan, H., Sivakumar, P., Chakrabarty, R., & Thomas, G. (2003). Scavenging of reactive oxygen species in NaCl-stressed rice (Oryza sativa L.)—differential response in salt-tolerant and sensitive varieties. Plant Science.165,1411–1418. https,//
Van Breusegem, F., & Dat, J. F. (2006). Reactive oxygen species in plant cell death. Plant Physiology.14, 384–390. https,//
Verma, S., & Mishra, S. N. (2005). Putrescine alleviation of growth in salt stressed Brassica juncea by inducing antioxidative defense system. Journal of Plant Physiology. 162,677-669. 10.1016/j.jplph.2004.08.008
Wang, M., Zhao, X., Xiao, Z., Yin, X., & Xing, T. (2016). A Wheat Superoxide Dismutase Gene TaSOD2 Enhances Salt Resistance through Modulating Redox Homeostasis by Promoting NADPH Oxidase Activity. Plant Molecular Biology. 91,115-30. 10.1007/s11103-016-0446-y
Williams, V., & Twine, S. (1960). Flame Photometric Method for Sodium Potassium and Calcium. In, K. Peach and M.V. Tracey (eds.), Modern Methods of Plant Analysis 3-5 pp, Springer-Verlag, Berlin, Germany.
Yoshida, S., Forne, D. A., Cock, J. H., & Gomez, K. A. (1976).  Laboratory manual for physiological studies of rice. 3rd end. International rice research Institute, Manila, Philipine, 67pp.
Zhu, J. K. (2001). Plant salt tolerance. Trends Plant Science. 6,66–71. https,//