Identification of markers associated with major physiological and yield component traits under moisture deficit stress conditions in preferred donor lines paves the way for marker-assisted selection (MAS). In the present study, a set of 183 backcross inbred lines (BILs) derived from the cross HD2733/2*C306 were genotyped using 35K Axiom genotyping array and SSR markers. The multi-trait, multi-location field phenotyping of BILs was done at three locations covering two major wheat growing zones of India, north-western plains zone (NWPZ) and central zone (CZ) under varying moisture regimes. A linkage map was constructed using 705 SNPs and 86 SSR polymorphic markers. A total of 43 genomic regions and QTL × QTL epistatic interactions were identified for 14 physiological and yield component traits, including NDVI, chlorophyll content, CT, CL, PH, GWPS, TGW and GY. Chromosomes 2A, 5D, 5A and 4B harbors greater number of QTLs for these traits. Seven Stable QTLs were identified across environment for DH (QDh.iari_6D), GWPS (QGWPS.iari_5B), PH (QPh.iari_4B-2, QPh.iari_4B-3) and NDVI (QNdvi1.iari_5D, QNdvi3.iari_5A). Nine genomic regions identified carrying major QTLs for CL, NDVI, RWC, FLA, PH, TGW and biomass explaining 10.32-28.35% of the phenotypic variance. The co-segregation of QTLs of physiological traits with yield component traits indicate the pleiotropic effects and their usefulness in the breeding programme. Our findings will be useful in dissecting genetic nature and marker-assisted selection for moisture deficit stress tolerance in wheat.