Yellow rust, caused by Puccinia striiformis, is one of the most devastating diseases in wheat. A synthetic by elite recombinant inbred line (RIL) population derived from a cross, Botnol/Aegilops squarrosa (666)//Kachu was evaluated for yellow rust resistance in two different environments in Mexico. The population was subjected to DArT-seq analysis for an in-depth genetic characterization. A major effect rust resistance QTL (QTL2A.K) explaining up to 45% phenotypic variance was found to be contributed by Kachee, an elite line of International Maize and Wheat Improvement Center (CIMMYT) Mexico. The QTL2A.K was found to be contributed by a segment of 2NS Chromosome of Triticum ventricosum translocated into the short arm of bread wheat chromosome 2A (QTL2A.K). The position of QTL2A.K was confirmed using T. ventricosum specific primer VENTRIUP-LN2. Identified genomic regions are being introgressed in to the popular but susceptible wheat varieties through marker-assisted breeding for enhancing yellow rust resistance.235-241