Pattern of antioxidant enzyme activities under drought stress and exogenous application of proline in sunflower

Document Type : Research Paper

Authors

1 Breeding for Abiotic Stresses Lab., Dept. of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Tabriz 5166616471, Iran.

2 Seed and Plant Improvement Institute, Karaj, Iran

3 Department of Plant Breeding and Biotechnology, Faculty of Agriculture,University of Tabriz, Tabriz, Iran.

Abstract

Background and objectives: Drought stress disturbs metabolic pathways in plant cells that leads to increase in reactive oxygen species (ROS) abundance. Plants have high efficient defensive system that prevents oxidative stress and can eliminate free radicals. Therefore high activity of antioxidant enzymes is essential for drought stress tolerance. Knowledge of antioxidant enzymes function and variation can be suitable marker to identification of morphological characteristics linked to this enzyme activities in plant breeding programs under drought conditions.
Materials and methods: In order to study the effect of drought stress and endogenous treatment of proline on the anti-oxidant enzymes superoxide dismutase (SOD), catalase (CAT), and peroxidase (POX), drought treatments (0, -4, -8 bar created by adding polyethylene glycol-6000) and 3 levels of proline (0,5, 10 mM) were applied on sunflower in seedling stage. This research was conducted in a factorial experiment based on completely randomized design, including 10 sunflower genotypes (in two susceptible and tolerant groups).
Results: Based on the changes in proline content and shoot dry weight, two inbred lines BGK 329 and RGK 221 were identified as the most susceptible and most sensitive among 10 inbred lines. Then, stripe pattern of enzymes was studied in 7.5 percent horizontal acrylamide gel electrophoresis. Quantitate evaluation of activity of observed isozymes and statistical analysis of the data indicated that there is a significant difference among the activity of four enzymes SOD1, SOD2, CAT1, and POX1, out of all 7 enzymes. No significant change was observed in activity of isozyme SOD1 with increasing water stress and proline levels. Interactions of genotype× proline was significant (p < 0.05) for POX1 and SOD2. Drought stress had significant effect on isozyme CAT1 and interaction of stress× genotype affected the activity of isozyme SOD2. While the activity of isozyme POX1 was significant at stress× proline interaction, the triple effect of triple stress× proline× genotype was significant (p < 0.01) in the activity of isozyme POX1. SOD2 activity was largely influenced by the genotype so that at different levels of stress and proline, some genotypes increased and some others showed decreased activity. The exogenous 5mM proline treatment reduced the activity of isozyme POX1 and CAT1 under stress. Data analysis based on two susceptible and tolerant sunflower groups under drought stress showed that only POX1 activity was significant between the two groups and therefore.
Conclusion: Proline as an important amino acid induces drought stress tolerance. An efficient antioxidant defensive system with inducible expression system to increase the accumulation of proline have a vital role in the tolerance of drought stress environments. Isozyme of POX could be suggested to identify drought tolerant drought genotypes in seedling growth stage.

Keywords


  1. Abogadallah, M.J. 2010. Antioxidative defense under salt stress. Plant Signal Behav. 5: 4. 369-374.
  2. Anderson, M.D., Prasad, T.K., and Stewart, C.R. 1995. Changes in isozyme profiles of catalase, peroxidase and glutathione reductase during acclimation to chilling in mesocotyls of maize seedling. J Plant Physiol. 109: 4. 1247-1257.
  3. Anwar Hossain, M., and Fujita, M. 2010. Evidence for a role of exogenous Glycine betaine and proline in antioxidant defense and methylglyoxal detoxification systems in mung bean seedlings under salt stress. Plant Mol. Biol. 16:1.19-28.
  4. Ashraf, M., and Foolad, M.R. 2007. Roles of glycinebetaine and proline in improving plant to abiotic stress tolerance. Environ. Exp. Bot. 59: 2. 206-216.
  5. Baloglu, M.C., Kavas, M., Aydin, G., Oktem, H.A., and Yucel, A.M. 2012. Antioxidative and   physiological responses of two sunflower cultivars under PEG mediated drought stress. Turk J Botany. 36: 6. 707-714.
  6. Bai, L.P., and Sui, F.G. 2006. Effect of soil drought stress on leaf water status, membrane permeability and enzymatic antioxidant system of maize. Pedosphere. 16: 3. 326-332.
  7. Blokhina, O., Virolainen, E., and Fagerstedt, K.V. 2003. Antioxidants, oxidative damage and oxygen deprivation stress. A review. Ann Bot. 91: 2. 179-194.
  8. Casano, L.M., Martin, M., and Sabater, B. 1994. Sensitivity of superoxide dismutase transcript levels and activities to oxidative stress is lower in mature-senescent than in young barley leaves. Plant Physiol. 106: 3. 1033-1039.
  9. Celina, M.L., Gabriela, M.P., and Simon, D. 2004. Drought and CAT gene expression in wheat. J. Exp. Bot. 56: 2. 417-423.
  10. Jiang, Y., and Huang, N. 2001. Drought and Heat stress injury to two cool-season turfggrasses in relation to antioxidant metabolism and lipid peroxidation. Crop Sci. 41: 2. 436-442.
  11.  Jung, S. 2004. Variation in antioxidant metabolism of young and mature leaves of (Arabidopsis thaliana).subjected to drought. Plant Sci. 166: 2. 459-466.    
  12.  Maribel, L., Dionisio, S., and Satoshi, T. 1998. Antioxidant responses of rice seedlings to            salinity stress. Plant Sci. 135: 1. 1-9.
  13.  Manivannan, P., AbdulJaleel, C., Sankar, B., Kishorekumar, A., Somasundaram, R., Lakshmanan, G. M.A., and Panneerselvam, R. 2007. Growth, biochemical modifications and proline metabolism in (Helianthus annuus L.). as induced by drought stress. Colloids Surf B Biointerfaces.59: 2. 141-149.
  14. Masoumi, H., Masoumi, M., Darvish, F., Daneshian, J., Nourmohammadi, G.H., and Habibi, D. 2010. Change in several antioxidant anzymes activity and seed yield by water deficit stress in soybean (Glycine max L.) cultivars, Notulae Botanicae Horti Agrobotanici. Cluj-Napoca. 38: 3. 50-59. (In Persian)
  15. Pan, Y., Jun Wu, L., and Liang Yu Z. 2006. Effect of salt and drought stress on antioxidant enzymes activities and SOD isoenzymes of liquorice (Glycyrrhiza uralensis Fisch), J. Plant Growth Regul. 49: 2. 157-165.
  16. Sairam, k., Chandrasekhar, V., and srivasta, G.C. 2001. Comparison of hexaploid and tetraploid wheat cultivars in their responses to water stress. Biologia Plantarum. 44: 1. 89-94.
  17. Sairam, K., Deshmuh, P.S., and Shukla, D.S. 1997. Tolerance of drought and temperature stress in relation to increased antioxidant enzyme activity in wheat. J. Agron. Crop Sci. 178: 3. 171-178.      
  18. Sayfzadeh, S., Habibi, D., and Fathollah, D.F. 2010. Response of antioxidant enzyme activities and root yield in sugar beet to drought stress. Int. J. Agric. Biol. 13: 3. 358-362. (In Persian)
  19. Sharma, P., and Dubey, R.S. 2005. Drought induces oxidative stress and enhances the activities of antioxidant enzymes in growing rice seedlings. J. Plant Growth Regul. 46: 3. 209-221.
  20. Sharp, R.E., and Boyer, J.S. 1986. Photosynthesis at low water potential in sunflower: lack of photoinhibitory effects. Plant Physiol. 82: 1. 90-95.     
  21. Singh, A.M.L., and Saini, R.K. 2004. Seed germination and seedling growth of citrus (Cytrus species) root stocks under different salinity regimes. J. Agri. Sci. 74: 5. 246-248.
  22. Soltis, D., and Soltis, P.S. 1990. Isozymes in Plant Biology. Dioscorides Press. Portland. USA. 97: 13. 7051-7057.
  23. Soshinkova, T.N., Radyukina, N.L., Korolkova, D.V., and Nosov, A.V. 2013. Proline and functioning of the antioxidant system in (thellungiella salsuginea) plants and cultured cells subjected to oxidative stress. Russ. J. Plant Physiol. 60: 1. 41-54.
  24. Stepien, P., and Klobus, G. 2005. Antioxidant defense in the leaves of C3 and C4 plants under salinity stress. Physiol. Plant. 125: 1. 31-40.
  25. Sun, C., Du, W., Cheng, X., Xu, X., Zhang, Y., Sun, D., and Shi, J. 2010. The effects of drought stress on the activity of acid phosphatase and its protective enzymes in pigweed leaves. Afr. J. Biotechnol. 9: 6. 825-833.
  26. Valizadeh, M., Mohayeji, M., Yasinzadeh, N., Nasrullazade, S., and Moghaddam, M. 2011. Genetic diversity of synthetic alfalfa generations and cultivars using tetrasomic inherited allozyme markers. J. Agri. Sci. 13: 425-430. (In Persian)
  27. Yong, T.L., Zongsuo, S., and Feng, D. 2006. Effect of water deficits on the activity of anti-oxidative enzymes and osmoregulation among three different genotypes of (Radix astragali) at seedling stage. Colloids Surf B Biointerfaces. 49: 1. 60-65.