Our objective was to determine the value of corn (Zea mays) inbred Oh516 as a source of resistance to Aspergillus ear rot and aflatoxin accumulation in grain. Types and magnitudes of gene action associated with resistance were determined with generation means analysis. Molecular markers associated with resistance were identified from BCP1S1 families developed from the cross of the susceptible inbred B73 and Oh516. In 2001 and 2002, B73 (P1), Oh516 (P2), and the F1, F2, F3, BCP1, BCP2, and BCP1S1 generations were evaluated for aflatoxin concentration in grain, percent bright greenish yellow fluorescence (BGYF), and severity of Aspergillus ear rot following inoculation in Urbana, IL. BCP1S1 families testcrossed with LH185 also were evaluated at three locations in 2002. Molecular marker-quantitative trait loci (QTL) associations with all traits were determined with single factor analysis of variance. Dominance gene action was associated with aflatoxin concentration in grain and percent BGYF. QTL associated with aflatoxin accumulation in grain were identified on chromosomes 2, 3, and 7 (bins 2.01 to 2.03, 2.08, 3.08, and 7.06). Alleles from inbred Oh516 on chromosomes 2, 3, and 7 should improve resistance of commercially used, B73-type inbreds.