Quantitative expression study of some key genes of durum wheat in the face of mild and severe drought stress

Document Type : Original Article

Authors

1 Department of Agronomy and Plant Breeding, Faculty of Agriculture, Ilam University, Ilam, Iran.

2 Dryland Agricultural Research Institute, Sararood Branch, Agricultural Research, Education and Extension (AREEO), Kermanshah, Iran.

3 Department of Plant breeding and Biotechnology, Ker. C., Islamic Azad University, Kermanshah, Iran.

Abstract

Introduction: Drought stress is one of the most important factors limiting the productivity of durum wheat (Triticum durum L.) in arid and semi-arid regions of the world, and understanding the response of genes at the molecular level is essential for the development of tolerant cultivars. Therefore, the present study aimed to investigate the relative expression of six key genes related to drought response, including TdDhn5 and Wdhn13 (dehydrin genes), TdAPX1 and TdCAT1 (antioxidant genes), and TdDRF1 and TdDREB1 (transcription factors), in seven durum wheat genotypes including (Zahab, G6, G8 and G9 as sensitive and genotypes G10, G12 and G14 as tolerant under no stress, mild, and severe conditions. The primary objective of the study was to identify genotype-based expression patterns and assess their potential as factors for enhancing drought tolerance.
Materials and methods: Wheat seeds were grown in greenhouse conditions in pots with a volume of approximately 2 liters containing 1.5 kg of field soil. Drought stress was applied at the three-leaf stage at two levels: mild and severe. Mild drought with biweekly irrigation and severe drought stress at the 3-leaf stage with complete cutting. Irrigation was induced according to the method described by Moradi et al. (2024). Each irrigation was 250 cc. After four weeks of exposure to stress, sampling was performed. The third fully opened leaf was harvested from three biological replicates of the pot for each genotype in each treatment, with each replicate consisting of five plants. Total RNA was extracted from the third leaves of the seedlings at the 3-4 leaf stage by the TRIzol method and after cDNA synthesis, gene expression was quantified by the technique using the Actin gene as a reference gene. Data were analyzed using ANOVA and the LSD test (p < 0.05).
Results: The results showed that the expression of the dehydrin genes TdDhn5 and TdWdhn13 increased significantly under mild and severe stress conditions (50% and 100%, respectively). In severe stress, the highest expression value was observed in the G14 genotype for TdDhn5 and in the G12 genotype for TdWdhn13. Antioxidant genes TdAPX1 and TdCAT1 were also upregulated under both mild and severe stress conditions, with the increase being greater in drought-tolerant genotypes. Notably, under severe stress, TdAPX1 expression was prominent in Genotype G9, and TdCAT1 expression was observed in genotype G12. Transcription factors TdDRF1 and TdDREB1 demonstrated their regulatory role, with increased expression under mild stress and a peak under severe stress, particularly in Genotypes G12 and G14.
Conclusion: This study clarified gene- and genotype-specific expression patterns in response to drought stress and identified G12 genotype as a prominent candidate for tolerance. The coordinated increase in dehydrin genes and transcription factors in response to severe stress confirms their potential as molecular markers in durum wheat breeding programs for drought tolerance. The need for field studies to validate these findings is recommended.

Keywords

Main Subjects

References

Close, T.J. 1997. Dehydrins: a commonalty in the response of plants to dehydration and low temperature. Physiologia Plantarum, 100(2), 291-296.
Dubouzet, J.G., Sakuma, Y., Ito, Y., Kasuga, M., Dubouzet, E.G., Miura, S., Seki, M., Shinozaki, K. & Yamaguchi Shinozaki, K. 2003. OsDREB genes in rice, Oryza sativa L., encode transcription activators that function in drought, high salt and cold responsive gene expression. The Plant Journal, 33(4), 751-763.
Eftekhari, A., Baghizadeh, A., Yaghoobi, M.M., & Abdolshahi, R. 2017. Differences in the drought stress response of DREB2 and CAT1 genes and evaluation of related physiological parameters in some bread wheat cultivars. Biotechnol Equipmwnt, 31, 709-716.
FAO. 2023. World Food and Agriculture – Statistical Yearbook. Rome: FAO.
Farooq, M., Hussain, M. & Siddique, K.H. 2014. Drought stress in wheat during flowering and grain-filling periods. Critical Reviews in Plant Sciences, 33(4), 331-349.
Feki, K., Tounsi, S., Kamoun, H., Al-Hashimi, A. & Brini, F. 2024. Decoding the role of durum wheat ascorbate peroxidase TdAPX7B-2 in abiotic stress response. Functional & Integrative Genomics, 24(6), 223.
Ghorbel, M., Zribi, I., Besbes, M., Bouali, N. & Brini, F. 2023. Catalase gene family in durum wheat: genome-wide analysis and expression profiling in response to multiple abiotic stress conditions. Plants, 12(14), 2720.
Hassan, N.M., El-Bastawisy, Z.M., El-Sayed, A.K., Ebeed, H.T. & Alla, M.M.N. 2015. Roles of dehydrin genes in wheat tolerance to drought stress. Journal of Advanced Research, 6(2), 179-188.
Hekimi, S., Noë, A., Branicky, R., & Wang, Y. 2018 Superoxide dismutases: Dual roles in controlling
ROS damage and regulating ROS signaling. Journal of Cell Biology, 217, 1915–1928.
Kovacs, D., Agoston, B. & Tompa, P. 2008. Disordered plant LEA proteins as molecular chaperones. Plant Signaling & Behavior, 3(9), 710-713.
Lata, C. & Prasad, M. 2011. Role of DREBs in regulation of abiotic stress responses in plants. Journal of Experimental Botany, 62(14), 4731-4748.
Latini, A., Cantale, C., Thiyagarajan, K., Ammar, K. & Galeffi, P. 2022. Expression analysis of the TdDRF1 gene in field-grown durum wheat under full and reduced irrigation. Genes, 13(3), 555.
Lesk, C., Rowhani, P. & Ramankutty, N. 2016. Influence of extreme weather disasters on global crop production. Nature, 529(7584), 84-87.
Livak, K.J. & Schmittgen, T.D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 25(4), 402-408.
Miller, G., Suzuki, N., Ciftci Yilmaz, S. & Mittler, R. 2010. Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant, Cell & Environment, 33(4), 453-467.
Mohammadi, R. & Amri, A. 2013. Phenotypic diversity and relationships among worldwide durum wheat (Triticum turgidum L. var. durum) germplasm collection under rainfed conditions of Iran. Crop and Pasture Science, 64(2), 87-99.
Moradi, M., Etminan, A., Mohammadi, R., Shooshtari, L., Mehrabi, A.M. 2024. Assessment of expression pattern of some antioxidant and dehydrin genes in durum wheat genotypes under water deficit conditions. Agricultural Biotechnology Journal, 16 (4), 189-208.
Noctor, G. & Foyer, C.H. 1998. Ascorbate and glutathione: keeping active oxygen under control. Annual Review of Plant Biology, 49(1), 249-279.
Ozturk, Z.N., Talamé, V., Deyholos, M., Michalowski, C.B., Galbraith, D.W., Gozukirmizi, N., Tuberosa, R. & Bohnert, H.J. 2002. Monitoring large-scale changes in transcript abundance in drought-and salt-stressed barley. Plant Molecular Biology, 48(5), 551-573.
Pantha, S., Kilian, B., Oezkan, H., Zeibig, F. & Frei, M. 2025. A comparison of drought responses in wild wheat relatives and domesticated wheat grown under irrigated and rainfed field conditions. Field Crops Research, 321, 109678.
Shinozaki, K. & Yamaguchi-Shinozaki, K. 2007. Gene networks involved in drought stress response and tolerance. Journal of Experimental Botany, 58(2), 221-227.
Smirnoff, N., 2000. Ascorbate biosynthesis and function in photoprotection. Philosophical transactions of the Royal Society of London. Series b: Biological Sciences, 355(1402), 1455-1464.
Souza, C.C., Oliveira, F.A., Silva, I.F., & Amorim Neto, M.S. 2000. Evaluation of methods of available
water determination and irrigation management in ‘‘terra roxa’’ under cotton crop. Revista Brasileira de Engenharia Agrícola e Ambiental, 4, 338-342

Files

Share

How to cite

Statistics