Study of antioxidant defense genes expression profile pattern of rice (Oryza sativa L.) cultivars in response to drought stress

Document Type: Research paper


1 Department of Plant Breeding and Biotechnology, Faculty of Plant Production, Gorgan University of Agricultural Science and Natural Resources, P. O. Box: 491381-5739, Gorgan, Iran.

2 Department of Plant Production, Faculty of Agriculture and Natural Resources, Gonbad Kavous University, Gonbad Kavous, Iran.

3 Nuclear Agriculture Group, Nuclear Science and Technology Research Institute, Karaj, Iran.



Drought stress is one of the important factors that restrict crop production in the world. This studywas conducted to investigate defense gene expression in response to drought stress, and also to evaluate the drought tolerance and its mechanism in rice cultivars based on randomized complete block design in two separate environments (drought stress and non-stress). The rice cultivars used included two commercial cultivars, i.e., Ahlemi-Tarom, Sepidrood, and three promising lines of fourth mutant generation called 4, 94 and 95 tested on the research farm at Gonbad Kavous University in 2018. For carrying out the drought stress-related experiment, irrigation was completely cut off 40 days after the transplantation (a stage with maximum tillering) until the end of the growth period. In seeding stage, each plant leaf was separately sampled in each block (under the stress and non-stress conditions). TBARM, superoxide andhydrogen peroxide contents and oxidative-stress related gene expression including superoxide dismutase, catalase, ascorbate peroxidase and glutathione peroxidase were measured. Results indicated that the drought-tolerant mutant lines had lower TBARM, superoxide andhydrogen peroxide contents compared to the studied cultivars. The mutant lines 94, 4 and 95 and Ahlemi-Tarom and Sepidrood had the highest to lowest levels of gene expression and yield in drought stress condition, sequentially. Accordingly, there was a complete correlation between the decreases in TBARM, superoxide andhydrogen peroxide and an increase in the level of gene expression. Mutant 94 had the highest yield and probably a suitable genetic potential in relation to drought tolerance. Therefore, the mentioned mutant is recommended for carrying out further studies.


Ahmed M., Hassen F. U., Qadeer U., and Aslam M. A. (2011). Silicon application and drought tolerance mechanism of sorghum. African Journal of Agricultural Research, 6: 594–607.

Alscher R. G., Ertrurk N., and Heath L. S. (2002). Role of superoxide dismutases (SODs) in controlling oxidative stress in plants. Journal of Experimental Botany, 53(372): 1331–1341.

Apel K., and Hirt H. (2004). Reactive oxygen species: metabolism, oxidative stress and signal transduction. Annual Review of Plant Biology, 55: 373–399.

Beyer W., Imlay J., and Fridovich I. (1991). Superoxide dismutases. Progress in Nucleic Acid Research and Molecular Biology, 40: 221–253.

Chaves M. M., Maroco J. P., and Pereira J. S. (2003). Understanding plant responses to drought from genes to the whole plant. Functional Plant Biology, 30(3): 239–264.

Chen Z., Chen B., Guo Q., Shi L., He M., Qin Z., Li L., He P., Wang Z., Hu D., and Yang S. (2015). A time-Course proteomic analysis of rice triggered by plant activator BTH. Journal of Plant Growth Regulation, 34: 392–409.

Doke N., Miura Y., Sanchez L., and Kawakita K. (1994). Involvement of superoxide in signal transduction: responses to attack by pathogens, physical and chemical shocks, and UV irradiation. In: Foyer CH, Mullineaux PM, editors. Causes of photo-oxidative stress and amelioration of defense systems in plants. Boca Raton, FL: CRC Press, 177–198.

Ebadi Almas D., Navabpour S., Yamchi A., Zaynali Nezhad K., Momeni A., and Mirzaghaderi G. (2019). Evaluation of several important defense enzyme activities in mutant rice resistant to blast disease. Journal of Nuclear Science and Technology, 17(86): 100–108. (in Persian)

Elstner E. F., and Heupel A. (1976). Inhibition of nitrite formation from hydroxyl ammonium chloride: a simple assay for superoxide dismutase. Analytical Biochemistry, 70(2): 616–620.

Ettinger W. F., and Harada J. J. (1990). Translational or post-translational processes affect differentially the accumulation of isocitrate lyase and malate synthase proteins and enzyme activities in embryos and seedlings of Brassica napus. Archives of Biochemistry and Biophysics, 281(1): 139–143.

Foyer C. H., and Noctor G. (2009). Redox regulation in photosynthetic organisms: signaling, acclimation, and practical implications. Antioxidants & Redox Signaling, 11(4): 861–905.

Gechev T., Gadjev I., Van Breusegem F., Inze D., Dukiandjiev S., Toneva V., and Minkov I. (2002). Hydrogen peroxide protects tobacco from oxidative stress by inducing a set of antioxidant enzymes. Cellular and Molecular Life Sciences, 59(4): 708–714.

Ghasemi Mesrami A., Navabpour S., Yamchi A., and Hooshmand S. (2016a).Effect of salinity stress on some morphological and biochemical characteristics of three bread wheat (Trticum aestivum L.) genotypes. Environmental Stresses in Crop Siences, 8(2): 273–283. (in Persian)

Ghasemi Mesrami A., Navabpour S., Yamchi A., and Hooshmand S. (2016b).Evaluation of genetic diversity in salinity tolerance of wheat mutant lines using ISSR molecular marker. Modern Genetics Journal,11(2): 271–279. (in Persian)

Gill S. S., and Tuteja N. (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry, 48(12): 909–930.

Golden T. A., Hinerfeld D., and Melov S. (2002). Oxidative stress and aging beyond correlation. Aging Cell, 122: 117–123.

Haddad R., and Kamangar A. (2015). The ameliorative effect of silicon and potassium on drought stressed grape (Vitis vinifera L.) leaves. Iranian Journal of Genetics and Plant Breeding, 4(2): 48–58.

Haddad R., Morris K., and Buchanan-Wollaston V. (2004). Expression analysis of two senescence involved genes in Brassica napus and Arabidopsis thaliana. Journal of Agricultural Science and Technology, 6(1-2): 63–72.

Hagege D., Nouvelot A., Boucard J., and Gaspar T. (1990). Malondialdehyde titration with thiobarbiturate in plant extracts: avoidance of pigment interference. Phytochemical Analysis, 1: 86–89.

Heidari R., and Haddad R. (2015).Silicon rule in molecular activity of barley antioxidant genes under drought stress. Crop Biotechnology, 9: 65–76. (in Persian)

Hodges D. M., Delong J. M., Forney C. F., and Prange R. K. (1999). Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation plant tissues containing anthocyanin and other interfering compounds. Planta, 207:604–611.

Hsu S. Y., and Kao C. H. (2003). Differential effect of sorbitol and polyethylene glycol on antioxidant enzymes in rice leaves. Plant Growth Regulation, 39(1): 83–90.

Kamali N., Navabpour S., Soltanloo H., and Kalate M. (2015).Changes in Metallothionein gene expression, chlorophyll content and some agronomic traits in response to salt stress in wheat. Journal of Crop Breeding, 7(15): 57–67.

Kar R. K. (2011). Plant responses to water stress: role of reactive oxygen species. Plant Signalling & Behavior, 6(11): 1741–1745.

Kazemi G., Navabpour S., and Ramezanpour S. S. (2012). Evaluation of gene Catalase expression and morphological traits under salinity stress in two susceptible and tolerant cultivars of bread wheat. Modern Genetics Journal,7(1): 79–87. (in Persian)

Kazerani B., and Navabpour S. (2019). Induced genes expression pattern in response to drought stress at seedling stage of wheat. Journal of Plant Physiology and Breeding, 9(1): 111–128.

Kazerani B., Navabpour S., Sabouri H., Ramezanpour S. S., Zaynali Nezhad K., and Eskandari, A. (2018). Determination of the best selection indices in mutant lines of rice at different moisture conditions. Crops Improvement, 20(1): 173–189. (in Persian)

Kazerani B., Navabpour S., Sabouri H., Ramezanpour S. S., Zaynali Nezhad K., and Eskandari, A. (2019a). Evaluation of proline content and enzymatic defense mechanism in response to drought stress in rice. Iranian Journal of Plant Physiology, 9(2): 2749–2758.

Kazerani B., Navabpour S., Sabouri H., Ramezanpour S. S., Zaynali Nezhad K., and Eskandari, A. (2019b). Evaluation and selection of rice mutant lines based on drought tolerance indices. Journal of Plant Production Research, 25(4): 15–31. (in Persian)

Kazerani B., Navabpour S., Sabouri H., Ramezanpour S. S., Zaynali Nezhad K., and Eskandari, A. (2019c). Evaluation of drought tolerance in rice recombinant inbred lines using stress tolerance indices. Seed and Plant Production Journal, 59–81. (in Persian)

Kiani S. P., Maury P., Sarrafi A., and Grieu P. (2008). QTL analysis of chlorophyll fluorescence parameters in sunflower (Helianthus annuus L.) under well-watered and water-stressed conditions. Plant Science, 175(4): 565–573.

Kim Y. H., Khan A. L., Waqas M., Shim J. K., Kim D. H., Lee K. Y., and Lee I. J. (2014). Silicon application to rice root zone influenced the phytohormonal and antioxidant responses under salinity stress. Journal of Plant Growth Regulation, 33(2): 137–149.

Kolupae Y., Karpets Y. V., and Kosakivska I. V. (2008). The importance of reactive oxygen species in the induction of plant resistance to heat stress. Plant Physiology, 34(3-4): 251–266.

Larkindale J. D., Hall J. R., Knight M., and Vierling E. (2005). Heat stress phenotypes of Arabidopsismutants implicate multiple signalling pathways in the acquisition of thermo-tolerance. Plant Physiology, 138: 882–897.

Leonowicz G., Trzebuniak K. F., Zimak-Piekarczyk P., Ślesak I., and Mysliwa-Kurdziel B. (2018). The activity of superoxide dismutases (SODs) at the early stages of wheat deetiolation. PLoS ONE, 13(3): e0194678.

Lewis N. G., and Yamamoto E. (1990). Lignin-Occurrence, biogenesis and biodegradation. Annual Review of Plant Physiology, 41: 455–496.

Loreto F., and Velikova V. (2001). Isoprene produced by leaves protects the photosynthetic apparatus against ozone damage, quenches ozone products, and reduces lipid peroxidation of cellular membranes. Plant Physiology, 127: 1781–1787.

Luna C. M., Pastori G. M., Driscoll S., Groten K., Bernard S., and Foyer C. H. (2004). Drought controls on H2O2 accumulation, catalase (CAT) activity and CAT gene expression in wheat. Journal of Experimental Botany, 56: 417–423. 

Mackerness S. A. H. (2000). Plant responses to ultraviolet-B (UV-B: 280-320 nm) stress: What are the key regulators? Invited review. Plant Growth Regulation, 32(1): 27–39.

Mazandarani A., Rahim Malek M., Navabpour S., and Ramezanpour S. (2014).Evaluation of chlorophyll content and genes expression (Catalase and DREB1) in soybean cultivars under drought stress condition. Agricultural Biotechnology, 5(1): 45–58. (in Persian)

Michalak A. (2006). Phenolic compounds and their antioxidant activity in plants growing under heavy metal stress. Polish Journal of Environmental Studies, 15(4): 523–530.

Mittler R., Vanderauwera S., Gollery M., and Van Breusegem F. (2004). Reactive oxygen gene network of plants. Trends in Plant Science, 9(10): 490–498.

Moller I. M., and Sweetlove L. J. (2010). ROS signalling-specificity is required. Trends in Plant Science, 15(7): 370–374.

Navabpour S. (2012a). Effect of abscisic acid application on enzymatic and nonenzymatic activity in barley (Hordeum vulgare L.). Cereal Research, 1(1): 39–51. (in Persian)

Navabpour S. (2012b). Molecular analysis of physiological stage of leaf senescence. Genetic Engineering and Biosafety Journal, 1(2): 63–72. (in Persian)

Navabpour S. (2012c).Study of in-situ Glutathion Peroxidase gene expression in growth stages of Brassica napus at drought stress condition. Journal of Plant Production, 19(3): 109–124. (in Persian)

Navabpour S. (2013). Induced genes expression pattern in response to drought stress in Rapeseed (Brassica napus). Seed and Plant Improvement Journal, 1-29(3):535–549. (in Persian)

Navabpour S., Bagherieh Najar M. B., and Haddad R. (2011). Comparison of induced gene response to stressful treatment in Spinacia oleracea and Brassica napus. Agricultural Biotechnology, 2(1): 1–10. (in Persian)

Navabpour S., and Mazandarani A. (2017). Molecular and biochemical evaluation of two bread wheat cultivars (Triticum aestivum L.) under oxidative stress. Cereal Research, 7(3): 357–367. (in Persian)

Navabpour S., Mirkarimi S., and Mazandarani A. (2013). Evaluation of enzymatic and non-enzymatic defense mechanism in response to Charcoal Rot disease during growth stage in soybean. Crop Biotechnology, 3(5): 63–73. (in Persian)

Navabpour S., Morris K., Harrison E., Makerness S., and Buchanan-Wollaston V. (2003). Expression of senescence-enhanced genes in response to oxidative stress. Journal of Experimental Botany, 54: 2285–2292.

Navabpour S., Ramezanpour S., and Kazemi G. (2015a).Molecular analysis of hypersensitive reaction and senescence process in wheat leaves. Modern Genetics Journal, 10(1): 59–68. (in Persian)

Navabpour S., Ramezanpour S., and Mazandarani A. (2016).Evaluation of enzymatic and non-enzymatic defense mechanism in response to drought stress during growth stage in soybean. Plant Production Technology, 7(2): 39–54. (in Persian)

Navabpour S., Ramezanpour S. S., Soltanloo H., and Vakili Bastam S. (2015b). Drought stress changed expression profile of some genes in tillering and pollination stages of adult wheat. Applied Science Reports, 9(2): 100–109.

Neill S. J., Desikan R., Clarke A., Hurst R., and Hancock J. T. (2001). Hydrogen peroxide and nitric oxide as signalling molecules in plants. Journal of Experimental Botany, 52: 9–17.

Noctor G., and Foyer C. H. (1998). Ascorbate and glutathione: keeping active oxygen under control. Annual Review of Plant Physiology and Plant Molecular Biology, 49: 249–279.

Pfaffl M. W., Horgan G. W., and Dempfle L. (2002). Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Research, 30(9): 1–10.

Rossatto T., Amaral M. N., Benitez L. C., Vighi I. L., Braga E. J. B., Junior A. M. M., Maia M. A. C., Pinto L. S. (2017). Gene expression and activity of antioxidant enzymes in rice plants, cv. BRS AG, under saline stress. Physiology and Molecular Biology of Plants, 23(4): 865–875.

Sabouri, H., Biabani, A., Fakhzri Moghaddam, A., Katouzi, M., and Ebadi, A. A. (2008a). Genetic analysis of agronomic and qualitative traits in Iranian rice. Final Report of Project, Gonbad Kavous University, Gonbad Kavous, pp. 48. (In Persian)

Sabouri, H., Biabani, A., Fakhzri Moghaddam, Mollashahi, M., Sabouri, A., and Katouzi, M. (2008b). Genetic analysis of agronomic traits in Iranian rice using diallel method. Final Report of Project, Gonbad Kavous University, Gonbad Kavous, pp. 49. (In Persian)

Sabouri, H., Sabouri, A., Jafarzadeh, M. R., Sajjadi, S.J., Mollashahi, M., and Jafarian, H. A. (2011). Introduction of tolerant rice cultivars for Gonbad Kavous region. Final report of project, Gonbad Kavous University, Gonbad Kavous, pp. 46. (In Persian)

Sharma P., Jha A. B., Dubey R. S., and Pessarakli M. (2012). Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. Journal of Botany, 2012: 1–26.

Shigeoka S., Ishikawa T., Tamoi M., Miyagawa Y., Takeda T., Yabuta Y., and Yoshimura K. (2002). Regulation and function of ascorbate peroxidase isoenzymes. Journal of Experimental Botany, 53(372): 1305–1319.

Ślesak I., Libik M., Karpinska B., Karpinski S., and Miszalski Z. (2007). The role of hydrogen peroxide in regulation of plant metabolism and cellular signalling in response to environmental stresses. Acta Biochimica Polonica, 54(1): 39–50.

Szymańska R., Ślesak I., Orzechowska A., and Kruk J. (2017). Physiological and biochemical responses to high light and temperature stress in plants. Environmental and Experimental Botany, 139: 165–177.

Velikova V., Yordanov I., and Edreva A. (2000). Oxidative stress and some antioxidant system in acid rain treated bean plants: protective role of exogenous polyammines. Plant Science, 151:59–66.

Xu K., Chen S., Li T., Ma X., Liang X., Ding X., Liu H., and Luo L. (2015). OsGRAS23, a rice GRAS transcription factor gene, is involved in drought stress response through regulating expression of stress-responsive genes. BMC Plant Biology, 15(14): 1–13.

Yoo Y., Nalini Chandran A. K., Park J., Gho Y., Lee S., An G., and Jung K. (2017). OsPhyB-Mediating novel regulatory pathway for drought tolerance in rice root identified by a global RNA-Seq transcriptome analysis of rice genes in response to water deficiencies. Frontiers in Plant Science, 8: 1–19.

Yoshida, S. (1981). Fundamentals of rice crop science. International Rice Research Institute, Los Banos, Laguna, Philippines, pp. 269.

Zhang J., Zhang S., Chang M., Jiang H., Zhang X., Peng C., Lu X., Zhang M., and Jin J. (2018). Effect of drought on agronomic traits of rice and wheat: A meta-analysis. International Journal of Environmental Research and Public Health, 15(5): 1–14.