Quantifying root structure response to multiple defoliation events in a grazing situation is critical in developing management plans for warm-season tallgrasses. A pasture experiment was conducted in 1999, 2000, and 2001 near Mead, NE. The objective of the experiment was to determine the effect of timing and frequency of grazing on big bluestem (Andropogon gerardii Vitman) etiolated tiller growth and root and rhizome structure. Paddocks were grazed at a stocking rate of 9.9 Animal Unit Month (AUM) ha-1 in two to four cycles from mid-May to early-September. In April 2002, five 6.6- X 132-cm soil cores were extracted from each paddock. Soil cores were subsampled at 30-cm depth increments for estimates of root mass, root surface area, and root volume. Etiolated tiller tents were used to estimate organic reserves of big bluestem in each paddock in spring 2002. Mean number and weight of etiolated tillers were reduced by up to 40% and 50%, respectively, in paddocks grazed in a sequence of June after internode elongation, early August, and early September. Root structure in the top 30 cm of the soil profile was affected most by multiple defoliation events with <40 d of recovery between grazing periods. Root mass decreased by 25%, while mean surface area and volume of roots declined 10 and 15%, respectively, in the upper 30 cm of the soil profile in paddocks grazed in the sequence of post-internode elongation in June, early-August, and early-September. To maintain vigorous big bluestem pastures, grazing management should concentrate on the elongation and postelongation periods. Grazing at the elongation stage should be rotated among paddocks in successive years and the recovery period following grazing at internode elongation should be >40 d.