Horseweed has rapidly become a major weed in soybean and cotton production fields of the United States, and Indiana farmers ranked horseweed as one of the five worst weeds in their fields during a mail survey in 2003. Glyphosate resistance in horseweed is conferred by a single, incompletely dominant gene. Horseweed populations possess a high level of variability in their response to glyphosate. Horseweed has also evolved resistance to acetolactate synthase (ALS) inhibitors, and biotypes resistant to ALS-inhibiting herbicides and glyphosate are in many of the same areas. An experiment was designed to determine whether glyphosate resistance can be transferred by pollen. We found glyphosate-resistant plants in 1.1 to 3.8% of the progeny. Segregation ratios fit the expected 3 : 1 resistant : sensitive ratios confirming that glyphosate resistance in horseweed can transfer to closely located glyphosate-susceptible biotypes under open-pollinated conditions at low frequencies. The hypothesis of a follow-up experiment was that first-generation progeny of parent plants, selected on a continuum of low to high phenotypic response to glyphosate, will inherit respective low to high phenotypic responses to glyphosate. The variability in field-collected populations (low-level to high-level glyphosate resistance) ranged from 2 to 14 times the commonly recommended field use rate of glyphosate. However, low- and high-levels of glyphosate resistance were not observed in first-generation progeny. We conclude that differential glyphosate responses observed among parental populations was due to different frequencies of the resistance allele within the populations, rather than the presence of different resistance alleles.