Anecdotal reports suggest waterhemp is becoming more difficult to control with glyphosate. Waterhemp accessions collected primarily in Illinois before 1996, in 1998 to 1999, and in 2003, were evaluated for differential responses to glyphosate treatment. The pre-1996 group consisted of 10 accessions, whereas the 1998 to 1999 and 2003 groups consisted of 45 accessions each. Plants 10 to 12 cm in height from each waterhemp accession were treated with 0 or 220 g ae ha−1 glyphosate. Although shoot dry biomass 14 d after treatment, expressed as a percentage of the untreated, varied within and among accessions, the mean responses of the three groups were similar. However, when glyphosate responses were divided into 10 arbitrary classes (0 to 10, 11 to 20, …, and 81 to 90; > 91% of untreated), the number of plants within each class differed among groups. Most notably, the proportion of plants in the range of 0 to 30% of the untreated control decreased with time when progressing from the pre-1996 to the 2003 accessions. The relative frequency of waterhemp plants that were ≥ 100% of the untreated plants (i.e., uninjured by glyphosate) in the pre-1996, 1998 to 1999, and 2003 groups were 5, 6, and 5%, respectively. In a separate trial, waterhemp populations that exhibited the least sensitivity to glyphosate at 220 g ha−1 were controlled by 870 g ha−1 glyphosate. These results suggest that waterhemp, as a whole, within Illinois has not become less sensitive to glyphosate over the past few years. However, the proportion of plants that are most sensitive to glyphosate may be decreasing within populations.