3 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture Drought is one of the major ecological factors limiting crop production and food quality globally, especially in the arid and semi-arid areas of the world. Among crop plants, wheat (Triticum aestivum), which often experiences water-limited conditions, is an attractive study system because of the natural genetic variation in traits related to drought tolerance (Loggini et al., 1999). 4 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture Extreme environmental conditions that induce oxidative stress have been associated to an increased carbonyl groups content and to an induction in protease activity. Objective: The objective of the present study was to investigate mechanisms responsible for acclimatization to drought in two wheat cultivars, Veery (drought tolerant) and Sids-1 (drought susceptible) subjected to two drought phases interrupted by 2d re-watering period. 6 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture 7/31/2017 For these purposes the glutathione system of the tissue and its relation to protein synthesis was investigated in both cultivars upon dehydration and rehydration. Also to understand the role of proteolysis in the response stress. 7 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture to drought 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture 8 7/31/2017 Two wheat (T. aestivum L.) cultivars, namely Veery (drought tolerant) and Sids-1 (drought susceptible) were used in this study for: Estimation of Relative Water Content (RWC) Determination of Oxidative stress indices (protein oxidation) Glutathione determination Glutathione reductase (GR) assay Measurement of azocaseinolytic activity 9 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture 7/31/2017 Cont. Sids AA Cv Sids 1 Cont. Veery * * * * 60 (%) Cv Veery * * 80 * * * * * * * 40 * 20 0 Carbonyl group content (%) Relative water content 100 200 B 150 * * * * * * * * * * * * * 100 * * 50 0 0 4 8 2 Rew First drought phase 4 8 12 Fig. (1): relative water content (%) (A) and Carbonyl group content (B) of well watered and drought stressed wheat cultivars CV Sids-1 (sensitive) and CV Veery (Tolerant) subjected to two drought phases interrupted by 2d rewatering. Each value represents the mean ±SE of five replicates. Cont. Sids-1 = Control of sensitive cultivar; cv Sids-1= drought stressed sensitive cultivar, Cont. Veery; Control of tolerant cultivar, Cv Veery; drought stressed tolerant cultivar, 2 Rew; 2 days rewatring. Significant differences (P<0.05) between treatments according to LSD test are shown by an asterisk. Second drought phase Treatments (d) Fig. (1 ): 11 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture 7/31/2017 Fig. (2) Changes in glutathione status, GSH, GSSG and GSG/GSSG ratio in wheat cultivars cv sids-1 and cv Veery exposed to two drought cycles interrupted by 2d re-watering (2Rew). Each value represents the mean ±SE of five replicates. Cont. Sids-1; Control of sensitive cultivar, cv Sids-1; drought stressed sensitive cultivar, Cont. Veery; Control of tolerant cultivar, Cv Veery; drought stressed tolerant cultivar, 2 Rew; 2 days re-watering. Significant differences (P<0.05) between treatments according to LSD test are shown by an asterisk. 12 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture 25 * * GR (U/mg protein) 20 * * 15 * * * * * * * 10 5 0 2 4 8 2 Rew First drought phase 4 8 12 * Fig. 3 : Glutathione reductase (GR) activity of well watered and drought stressed wheat cultivars CvSids-1 and Cv Veery subjected to two drought phases interrupted by 2d re-watering. Each value represents the mean ±SE of five replicates. Cont. Sids-1; Control of sensitive cultivar, cv Sids-1; drought stressed sensitive cultivar, Cont. Veery; Control of tolerant cultivar, Cv Veery; drought stressed tolerant cultivar, 2 Rew; 2 days rewatring. Significant differences (P<0.05) between treatments according to LSD test are shown by an asterisk. Second drought phase Treatments (d) 13 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture Cont. Sids 100 Cv Sids 1 Cont. Veery A 80 * * Total soluble protein mg g-1 DW A Azocasinolytic activity unitsmg -1protein h-1 Cv Veery 60 * * * * * * * 40 * 20 0 10 8 6 4 B * * * * * * * * * * * * * * * * * * * * * * * 2 0 2 4 8 First drought phase 2 Rew Fig. (4): Total protein content (A) and Azocaseinolytic activity (B) of well watered and drought stressed wheat cultivars CV Sids-1 (sensitive) and CV Veery (tolerant) subjected to two drought phases interrupted by 2d re-watering. Each value represents the mean ±SE of five replicates. Cont. Sids-1; Control of sensitive cultivar, cv Sids-1; drought stressed sensitive cultivar, Cont. Veery; Control of tolerant cultivar, Cv Veery; drought stressed tolerant cultivar, 2 Rew; 2 days rewatring. Significant differences (P<0.05) between treatments according to LSD test are shown by an asterisk. 4 8 12 Second drought phase Treatments (d) 2nd International e-Conference on Agricultural Biosciences 2009 website: http://www.e-conference.elewa.org/agriculture 14 protein content (A)Conference and Azocasinolytic activity (B) of well watered and Fig. (3): Total drought stressed wheat cltivars CV sids (sensitive) and CV veery ( Tolerant) subjected to 15 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture A significant increase of carbonyl concentration in response to drought stress in both cultivars tested and after prolonged drought stress, indicating the presence of oxidized protein which are selectively recognized and degraded by proteolytic enzymes (Fig.1B) The ratio of GSH to GSSG was directly related to the level of drought stress, however, after 2d re-watering, the acclimatized wheat leaves for both cultivars showed higher values at the end of the second drought phase when compared to the results obtained after the completion of the first drought phase (Fig. 2C). 16 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture 7/31/2017 The high activity of GR, especially during the first drought phase, maintained glutathione pool in its reduced status. This is confirmed by the different behavior of cultivars, in which a relative low GR activity, also at high degree of drought, was associated with low GSH and high GSSG levels (Fig. 2 A, B and Fig. 3). 17 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture Lower azocaseinolytic activity was referred to cultivar type in order of increasing drought stress and associated with a distinct reduction of soluble protein content in the wheat leaves. Drought has been associated with decreased protein levels, increased proteolytic activity, and the up-regulation of cysteine protease genes (reviewed in Beers et al., 2004). 18 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture The recent literature confirms a central role of glutathione metabolism in plant responses to environmental stress. The results showed that antioxidant protection in both cultivars are mainly due to the enzymes and molecules involved in glutathione metabolism and re-watering regulates the timing of drought effect in part by delaying the increased proteolytic activity and subsequent protein 20 degradation. 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture Genetic, Physiological and Biochemical responses of some plants to interactive biotic and abiotic stress. Effect of some cumulative stress on the ultrastructure and antioxidant isozymes of some economically important plants. 21 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture 7/31/2017 This research was supported through the research center of Teachers College at King Saud University, project no. 12/1427. The author would also like to thank Dr. Salah Barakat, Professor of Plant Physiology, Botany Dept., Faculty of Science, Alexandria University for his comments and revision on the manuscript. 22 2nd International e-Conference on Agricultural Biosciences 2009 Conference website: http://www.e-conference.elewa.org/agriculture 7/31/2017
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