Transgenic maize, Zea mays L., expressing the Bacillus thuringiensis (Bt) Cry1Ab toxin has been planted to extensive areas across the United States and several other countries, but no resistance has been documented in field populations of lepidopteran target pests. This article describes the first report of resistance alleles to commercially available Cry1Ab Bt maize in a Louisiana population of sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae). Two hundred thirteen two-parent isolines of D. saccharalis were screened for Cry1Ab resistance on Bt maize leaf tissue using an F₂ screening technique. Larvae representing three isolines survived >15 d on Bt tissue in the F₂ generation. The second generation backcross progeny (B₁F₂) derived from isoline 52 completed larval development on Bt maize in the greenhouse. Segregation and resistance frequency analysis associated with isoline 52 suggested that Bt resistance is probably determined by a nearly completely recessive allele at a single locus. With this assumption, the estimated resistance allele frequency in this population is 0.0023, within a 95% confidence interval of 0.0003–0.0064.