Triticeae Genomics and Genetics, 2025, Vol.16, No.2, 63-71 http://cropscipublisher.com/index.php/tgg 69 8.3 Role of CRISPR and gene editing in validating candidate genes CRISPR/Cas9 and other gene editing techniques provide very direct tools for verifying the candidate genes identified by GWAS (Kumar et al., 2024). By site-specific modification or even knockout of a certain gene, researchers can clarify its role in salt tolerance and thereby create new allelic variations, preparing for breeding applications. In this way, the achievements of GWAS do not have to remain at the theoretical stage but can enter the actual genetic improvement process more quickly, and also open up a direct path for breeding barley strains with stronger salt tolerance. 9 Concluding Remarks In salt tolerance studies, GWAS has identified many important SNPS and candidate genes related to Na+/K+ balance, ionic homeostasis and antioxidant activity. Key loci like HKT1;5, as well as genes involved in ion transport, signaling, and stress response, are located on different chromosomes, and these regions are also considered "hotspots" for salt tolerance. When GWAS WAS combined with transcriptome analysis, the roles of these candidate genes were further verified, and their physiological functions in response to salt stress in plants also became clearer. These genetic markers are not only research achievements in the laboratory but can also directly enter the breeding process. They provide available tools for marker-assisted selection and genomic prediction, enabling breeders to more specifically introduce favorable alleles into favorable backgrounds, thereby enhancing yield stability and stress resistance under salt stress. Meanwhile, the stress memory of intergenerational and even cross-generational ones also provides new ideas for long-term adaptive breeding. For future salt-tolerant barley breeding to truly achieve breakthroughs, it is likely to rely on the coordination of several methods - GWAS, high-throughput phenotypic analysis, transcriptomics, and more advanced genomic selection models. Gene editing technologies like CRISPR will also come into play. They can not only verify the functions of key genes but also precisely modify them. When data-driven methods become more mature, the breeding speed of salt-tolerant varieties is expected to accelerate significantly. By then, these varieties will not only be able to maintain stable yields under conditions of increasing salinization and frequent climate change, but also provide solid support for global food security. Acknowledgments We express the profound sense of reverence to the entire research team, friends, and any other person who contributed; we have deep gratitude for you so much. Conflict of Interest Disclosure The authors affirm that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest. References Abdelrady W., Ma Z., Elshawy E., Wang L., Askri S., Ibrahim Z., Dennis E., Kanwal F., Zeng F., and Shamsi I., 2024, Physiological and biochemical mechanisms of salt tolerance in barley under salinity stress, Plant Stress, 11: 100403. https://doi.org/10.1016/j.stress.2024.100403 Allel D., Ben-Amar A., Badri M., and Abdelly C., 2016, Salt tolerance in barley originating from harsh environment of North Africa, Australian Journal of Crop Science, 10: 438-451. https://doi.org/10.21475/AJCS.2016.10.04.P6663X Allel D., Ben-Amar A., Badri M., and Abdelly C., 2019, Evaluation of salinity tolerance indices in North African barley accessions at reproductive stage, Czech Journal of Genetics and Plant Breeding, 55(2): 61-69. https://doi.org/10.17221/50/2017-CJGPB Alqudah A., Elkelish A., Alammari B., Alsubeie M., Hamed S., and Thabet S., 2024, Genetic mapping determining the key genomic Loci/QTNs for stress resilience via controlling antioxidant defenses in barley under salt stress, Plant Molecular Biology Reporter, 43: 324-335. https://doi.org/10.1007/s11105-024-01488-9 Boussora F., Triki T., Bennani L., Bagues M., Ali S., Ferchichi A., Ngaz K., and Guasmi F., 2024, Mineral accumulation, relative water content and gas exchange are the main physiological regulating mechanisms to cope with salt stress in barley, Scientific Reports, 14: 14931. https://doi.org/10.1038/s41598-024-65967-5
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