Present study aimed at molecular and phenotypic characterization of F3 population of a cross between leaf rust susceptible but well adapted bread wheat variety DWR 162 and resistant variety PBW 343 carrying highly effective alien genes Lr24 and Lr28 to identify recombinant lines with high productivity and leaf rust resistance. Fifty nine F3 lines with seven checks were evaluated for yield and yield contributing traits in randomized block design (RBD) with two replications. These lines were also screened against most virulent pathotypes of leaf rust 77-5 (121R63-1) in rust screening nursery. SCAR markers, namely SCS1302607 and SCS421570 linked to Lr24 and Lr28 genes facilitated identification of individual lines possessing the targeted genes. To identify the lines that mostly resemble ‘DWR 162’ variety, DUS (Distinctiveness Uniformity Stability) characterization was done. Nine lines were identified in F3 generation, possessing high yield potential and leaf rust resistance genes, Lr24 and Lr28 with the desirable features of DWR 162. Availability of a combination of two major rust resistance genes would facilitate the breeding program for developing new leaf rust resistant and high yielding cultivars.

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