Root growth is important to the competitive ability of plants, and understanding how herbage defoliation affects root growth has implications for development of management strategies. Objectives were to determine the effects of defoliation intensity and frequency on root characteristics and herbage production of slender wheatgrass (Elymus trachycaulus [Link.] Shinners), Nebraska sedge (Carex nebrascensis C. Dewey), and “Steadfast” birdsfoot trefoil (Lotus corniculatus L.). Plants of each species were transplanted into containers that had been placed in the ground at wet meadow field sites the prior year. There were eight replications of a control and five defoliation treatments, which were combinations of different frequencies (two or five times) and intensities (light or heavy) and haying. Treatments were applied for a single growing season, and aboveground biomass was collected. Containers were extracted in October, and plant crowns, rhizomes, and roots were separated from the soil. Defoliation treatment did not affect total root weight, length, and surface area of Nebraska sedge or birdsfoot trefoil (P > 0.10). Slender wheatgrass total root weight was less when defoliated five times (4.46 g · container⁻¹) than when defoliated twice (6.62 g · container⁻¹) during the growing season. More frequent defoliation of slender wheatgrass also reduced length (20%) and surface area (21%) compared to less frequent defoliation. However, defoliation frequency did not affect aboveground biomass. Defoliation intensity did not affect aboveground production or root characteristics of the three species. Abundant soil moisture in meadows likely buffers negative effects of defoliation. For all species, two defoliation events (e.g., haying followed by grazing) does not appear to negatively affect root growth and herbage production.