Evolution of TILLING as a Useful Tool for Durum Wheat Functional Studies and Breeding Programs

The development of improved crop varieties remains a significant challenge in agriculture, especially in light of climate change, increasing world population, and regional conflicts. To address these challenges, it is crucial to understand the genetic basis of desirable traits and harness genetic variability through breeding programs. Various strategies, including traditional breeding methods and modern biotechnological approaches, have been employed to generate genetic diversity in crops. Among these approaches, Targeting Induced Local Lesions IN Genomes (TILLING) is a reverse genetic tool that combines the use of mutagenesis and molecular techniques for the identification of point mutations in target genes. TILLING has been successfully applied to various plant species, facilitating the creation of novel allelic series and the study of gene function. In durum wheat, TILLING has been widely used to improve yield and nutritional value and give biotic and abiotic stress resilience. In addition, the TILLING platform has recently been employed to investigate promoter mutations and their effects on gene expression and phenotype modulation. Methodological advances, such as next-generation sequencing (NGS), have improved mutation discovery and reduced costs, making this approach accessible and efficient for durum wheat breeding programs. In conclusion, TILLING is a valuable tool for durum wheat improvement, offering insights into gene function, generating genetic diversity, and addressing challenges in agricultural sustainability. Continued research and technological innovations in TILLING hold promise for further enhancing crop resilience, nutritional quality, and yield in durum wheat and other major crops.