Experiments were conducted in the North Carolina State University Phytotron greenhouse and field locations in Clayton, Rocky Mount, and Lewiston-Woodville, NC, in 2002 to determine the effect of glyphosate on pollen viability and seed set in glyphosate-resistant (GR) corn. Varieties representing both currently commercial GR corn events, GA21 and NK603, were used in phytotron and field studies. All glyphosate treatments were applied at 1.12 kg ai ha−1 at various growth stages. Regardless of hybrid, pollen viability was reduced in phytotron and field studies with glyphosate treatments applied at the V6 stage or later. Scanning electron microscopy of pollen from affected treatments showed distinct morphological alterations correlating with reduced pollen viability as determined by Alexander stain. Transmission electron microscopy showed pollen anatomy alterations including large vacuoles and lower starch accumulation with these same glyphosate treatments. Although pollen viability and pollen production were reduced in glyphosate treatments after V6, no effect on kernel set or yield was found among any of the reciprocal crosses in the phytotron or field studies. There were also no yield differences among any of the hand self-pollinated (nontreated male × nontreated female, etc.) crosses. Using enzyme-linked immunosorbent assay to examine CP4-5-enolpyruvlshikimate-3-phosphate synthase expression in DKC 64-10RR (NK603) at anthesis, we found the highest expression in pollen with progressively less in brace roots, ear leaf, anthers, roots, ovaries, silks, stem, flag leaf, and husk.