RUDN Journal of Agronomy and Animal IndustriesRUDN Journal of Agronomy and Animal IndustriesВестник Российского университета дружбы народов. Серия: Агрономия и животноводство2312-797X2312-7988Peoples’ Friendship University of Russia named after Patrice Lumumba2030710.22363/2312-797X-2026-21-1-41-54DZOGIDGenetics and plant breedingГенетика и селекция растенийResearch ArticleIdentification of drought-tolerant bread wheat Triticum aestivum L. genetic resources using molecular markersИдентификация генетических ресурсов засухоустойчивоcти продовольственной пшеницы Triticum aestivum L. с использованием молекулярных маркеровhttps://orcid.org/0000-0002-4884-6481WalliMalek H.ВаллиМалек Хубаиш

Ph.D Student of Department of Agrobiotechnology, Agrarian and Technological Institute, RUDN University; Department of Biology, College of Nursing, Al-Muthanna University

аспирант агробиотехнологического департамента, аграрно-технологический институт, Российский университет дружбы народов; кафедра биологии, Колледж сестринского дела, Университет Аль-Мутанна

malek_h88@yahoo.comhttps://orcid.org/0000-0002-7353-78169937-6393DuksiFatimaДуксиФатима

Ph.D Student of Department of Agrobiotechnology, Agrarian and Technological Institute

аспирант агробиотехнологического департамента, аграрно-технологический институт

f.duksi@gmail.comhttps://orcid.org/0009-0005-6908-5385Al-jubouriZinaАль-ДжабориЗина

lecturer Department of Statistics, Faculty of Pharmacy

лектор кафедры статистики, фармацевтический факультет

Zinah480@gmail.comhttps://orcid.org/0000-0002-5208-0861ZargarMeisamЗаргарМейсам

PhD in Agricultural sciences, professor, Department of Agrobiotechnology, Agrarian and Technological Institute

доктор сельскохозяйственных наук, профессор агробиотехнологического департамента, аграрно-технологический институт

zargar-m@rudn.ruhttps://orcid.org/0000-0003-2817-8807AlhasnawiArshadАльхаснавиАршад

Ph.D., Department of Biology, College of education for pure Sciences

доктор наук кафедра биологии, Педагогический колледж чистых наук

arshad@mu.edu.iqRUDN UniversityРоссийский университет дружбы народовAl-Muthanna UniversityУниверситет Aль-Mутанна09052026211415411052026Copyright ©; 2026, Walli M.H., Duksi F., Al-jubouri Z., Zargar M., Alhasnawi A.Copyright ©; 2026, Валли М.Х., Дукси Ф., Аль-Джабори З., Заргар М., Альхаснави А.2026Walli M.H., Duksi F., Al-jubouri Z., Zargar M., Alhasnawi A.Валли М.Х., Дукси Ф., Аль-Джабори З., Заргар М., Альхаснави А.https://creativecommons.org/licenses/by-nc/4.0https://agrojournal.rudn.ru/agronomy/article/view/20307

To mitigate climate change and support the global agricultural economy, plant breeding and improvement programmes have received significant attention to obtain genetic resources rich in tolerant genes. In our study, twenty ISSR primers were used to evaluate genetic variation for 23 genotypes. SSR markers and the Sanger sequencing method were used to detect drought-tolerant genotypes and identify target gene loci. ISSR-PCR results showed a total of 820 DNA bands, of which 172 bands were polymorphic (117 non-unique bands and 55 unique bands) with a polymorphism percentage of 88.6. In the similarity matrix and dendrogram, genotypes were divided into clusters according to the genetic origins of the parents. These results demonstrated that ISSR markers are a valuable method for determining genetic variation, and identification of wheat genetic origins. Using primers SSR-PCR (Malek 1, Malek 2), genotypes (AB, NF, CL, UD, SA, TB) were detected as having drought tolerance genes. DNA sequencing by Sanger sequencing method of the genotypes, genes (DRF1, NAC20L) were identified in the wild parent (Triticum turgidum L), which is considered the source of the tolerant genes. Sample sequences and genotypes were recorded in the Gene Bank and The NCBI Bankit platforms. DNA sequencing technology has proven its effectiveness in confirming field results and identifying targeted genetic sites in the wheat genome.

Для смягчения последствий изменения климата и поддержки мировой аграрной экономики значительное внимание уделяется программам селекции и улучшения растений с целью получения генетических ресурсов, богатых генами засухоустойчивости. Оценку генетической изменчивости 23  генотипов проводили с использованием двадцати ISSR-праймеров. Выявляли засухоустойчивые генотипы и осуществляли идентификацию целевых локусов генов примененеим SSR-маркеров и метода секвенирования по Сэнгеру. Результаты ISSR-ПЦР показали в общей сложности 820 полос ДНК, из которых 172 полосы были полиморфными (117 неуникальных полос и 55 уникальных полос) с процентом полиморфизма — 88,6. В матрице сходства и дендрограмме генотипы были разделены на кластеры в соответствии с генетическим происхождением материнских форм. Полученные результаты продемонстрировали, что ISSR-маркеры являются ценным методом определения генетической изменчивости и идентификации генетического происхождения пшеницы. С помощью праймеров SSR-PCR Malek 1, Malek 2 выявили гены засухоустойчивости у генотипов AB, NF, CL, UD, SA, TB. Секвенирование ДНК генотипов методом Сэнгера позволило идентифицировать гены DRF1 и NAC20L у дикого родительского растения (Triticum turgidum L.), которое считается источником генов засухоустойчивости. Образцы последовательностей и генотипов зарегистрировали в Gene Bank и на платформах NCBI Bankit. Технология секвенирования ДНК доказала свою эффективность в подтверждении результатов полевых исследований и идентификации целевых генетических участков в геноме пшеницы.

drought stressISSRSSRDNA sequencingSanger sequencingзасухаISSRSSRсеквенирование ДНКгенетикасеквенирование по СэнгеруQureshi AS, Al-Falahi AA. Extent, Characterization and causes of soil salinity in Central and Southern Iraq and possible reclamation strategies. International Journal of Engineering Reseach and Applications. 2015;5(1).Al-Jassim KAH. Natural reasons causing soil salinity and its impact of plant production in Ali-Algharbi district. Iraqi Journal of Desert Studies. 2021;11(2):164–186. doi: 10.36531/desert.2022.172743 EDN: DODRDXBraun HJ, Atlin G, Payne T. Multi-location testing as a tool to identify plant response to global climate change. In: Climate change and crop production. Wallingford, UK: CABI; 2010. p.115–138. doi: 10.1079/9781845936334.0115Walli MH, AL-Jubouri Z, Madumarov MM, Margaryta M, Aldibe AAA. Genetic and environment diversity to improve wheat (Triticum spp.) productivity: a review. Research on Crops. 2022;23(2):295–306. doi: 10.31830/2348-7542.2022.041 EDN: ATEWCGJha S. Transgenic approaches for enhancement of salinity stress tolerance in plants. In: Singh SP, Upadhyay SK, Pandey A, Kumar S. (eds.) Molecular Approaches in Plant Biology and Environmental Challenges. Singapore: Springer; 2019. p. 265–322. doi: 10.1007/978-981-15-0690-1_14 EDN: DWRDUXJabari M, Golparvar A, Sorkhilalehloo B, Shams M. Investigation of genetic diversity of Iranian wild relatives of bread wheat using ISSR and SSR markers. Journal of Genetic Engineering and Biotechnology. 2023;21(1):73. doi: 10.1186/s43141-023-00526-5 EDN: JZLXAKAslanparviz M, Rashidi V, Omidi M, Etminan A, Ahmadzadeh A. Evaluation of population structure and estimation of genetic parameters in breeding lines and landraces populations of durum wheat using ISSR markers. Plant Genetic Researches. 2022;8(2):23–32. doi: 10.52547/pgr.8.2.2 EDN: IGJEZIAtsbeha G, Tesfaye K, Mekonnen T, Haileselassie T, Kebede M. Genetic diversity and population structure analysis of bread wheat (Triticum aestivum L.) germplasms as revealed by ISSR markers. Genetic Resources and Crop Evolution. 2024;71(6):2721–2735. doi: 10.1007/s10722-023-01791-6Shaheenuzzamn M, Liu T, Shi Sh, An P, Wu H, Wang Zh. Development of sequencing technology and role of next generation sequencing technologies in wheat research: a review. Pakistan Journal of Botany. 2020;52(5):1867–1878. doi: 10.30848/PJB2020-5(33) EDN: FGGOARDellaporta SL, Wood J, Hicks JB. A plant DNA minipreparation: Version II. Plant Molecular Biology Reporter. 1983;1(4):19–21. doi: 10.1007/BF02712670 EDN: RKVWRQPorebski S, Bailey LG, Baum BR. Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Molecular Biology Reporter. 1997;15(1):8–15. doi: 10.1007/BF02772108 EDN: XSNALFMandal AM, Alhasnawi A, Jasim H, Mohamad A. Evaluation of salt stress and molecular analysis of genetic variation of Iraqi rice cultivars. Biodiversitas. 2019;20(11):3309–3314. doi: 10.13057/biodiv/d201125 EDN: VIEVRBMohamad A, Alhasnawi AN, Kadhimi AA, Isahak A, Wan Yusoff WM, Che Radziah CMZ. DNA Isolation and Optimization of ISSR-PCR Reaction System in Oryza sativa L. International Journal on Advanced Science, Engineering and Information Technology. 2017;7(6):2264. doi: 10.18517/ijaseit.7.6.1621Alhasnawi AN. β-glucan-mediated alleviation of NaCl stress in Ocimum basilicum L. in relation to the response of antioxidant enzymes and assessment DNA marker. Journal of Ecological Engineering. 2019;20(8):90–99.doi: 10.12911/22998993/110790Chesnokov YuV, Artemyeva AM. Evaluation of the measure of polymorphism information of genetic diversity. Agricultural Biology. 2015;50(5):571–578. doi: 10.15389/agrobiology.2015.5.571eng EDN: UXSRIXTonk FA, Tosun M, Ilker E, Istipliler D, Tatar O. Evaluation and comparison of ISSR and RAPD Markers for assessment of genetic diversity in triticale genotypes. Bulgarian Journal of Agricultural Science. 2014;20(6):1413–1420.Sneath PHA, Sokal RR. Numerical Taxonomy: The Principles and Practice of Numerical Classification. W.H. Freeman; 1973.Niu X, Luo T, Zhao H, Su Y, Ji W, Li H. Identification of wheat DREB genes and functional characterization of TaDREB3 in response to abiotic stresses. Gene. 2020;740:144514. doi: 10.1016/j.gene.2020.144514 EDN: YFERCVHaque MS, Saha NR, Islam MT, Islam MM, Kwon SJ, Roy SK, WOO SH. Screening for drought tolerance in wheat genotypes by morphological and SSR markers. Journal of Crop Science and Biotechnology. 2021;24(1):27–39. doi: 10.1007/s12892-020-00036-7 EDN: YNFVMQThungo Z, Shimelis H, Odindo A, Mashilo J, Shayanowako A. Genetic relationship among selected heat and drought tolerant bread wheat genotypes using SSR markers, agronomic traits and grain protein content. Acta Agriculturae Scandinavica, Section B — Soil & Plant Science. 2020;70(7):594–604. doi: 10.1080/09064710.2020.1818818 EDN: ERXVWZVaja KN, Gajera HP, Katakpara ZA, Patel SV, Golakiya BA. Microsatellite markers based genetic diversity analysis for salt tolerance in wheat genotypes. Indian Journal of Agricultural Biochemistry. 2016;29(2):140–145. doi: 10.5958/0974-4479.2016.00023.XPeng J, Sun D, Nevo E. Wild emmer wheat, ‘Triticum dicoccoides’, occupies a pivotal position in wheat domestication process. Australian Journal of Crop Science. 2011;5(9):1127–1143.Breseghello F, Sorrells ME. Association mapping of kernel size and milling quality in wheat (Triticum aestivum L.) cultivars. Genetics. 2006;172(2):1165–1177. doi: 10.1534/genetics.105.044586Balter M. Seeking agriculture’s ancient roots. Science. 2007;316(5833):1830–1835. doi: 10.1126/science.316.5833.1830Börner A, Korzun V, Worland AJ. Comparative genetic mapping of loci affecting plant height and development in cereals. Euphytica. 1998;100(1–3):245–248. doi: 10.1023/A:1018364425150 EDN: XOCJIDBolot S, Abrouk M, Masood-Quraishi U, Stein N, Messing J, Feuillet C, Salce J. The ‘inner circle’ of the cereal genomes. Current Opinion in Plant Biology. 2009;12(2):119–125. doi: 10.1016/j.pbi.2008.10.011Buckler ES, Thornsberry JM, Kresovich S. Molecular diversity, structure and domestication of grasses. Genetics Research. 2001;77(3):213–218. doi: 10.1017/S0016672301005158 EDN: FOCLGL