Knowledge about how the type, timing, and arrangement of cultural practices influence weed species composition is important for understanding the ecological results of control strategies and designing alternative crop management systems. We evaluated weed seed density, diversity, and community composition from 1997 to 1999 in a 35-yr-old study comparing cropping sequences (continuous corn, corn–soybean, corn–oat–hay) and tillage systems (conventional, minimum, and no-tillage) in Wooster, OH. Weed seedbank diversity, as measured by species richness (S), evenness (J), and the Shannon–Weiner index (H′), was influenced by crop diversity; mean values for each of the indices were generally higher for all combinations of the three-crop sequence than for the corn monoculture or the corn–soybean rotation. Except for 1998, mean seed density (to a depth of 10 cm) was higher in continuous corn (x̄ = 5,220 m−2) than in corn and soybean rotations (x̄ = 2,360 m−2). Species richness and seed density were also affected by tillage. Mean values for S (x̄ = 10.8 species) and mean germinable seeds (x̄ = 5,960 m−2) were greatest in the no-tillage system, where the soil was disturbed only by the coulter units of the planter. Differences in weed seedbank community composition among tillage and rotation treatments were examined using two multivariate analyses. Using a multiresponse permutation procedure and canonical discriminant analysis, results suggest that the weed seed community in a corn–oat–hay rotational system differs in structure and composition from communities associated with continuous corn and corn–soybean systems. Additionally, germinable weed seed communities in no-tillage differed in composition from those in conventional and minimum tillage. Crop sequence and tillage system influenced weed species density and diversity and therefore community structure. Manipulation of these factors could help reduce the negative impact of weeds on crop production.Nomenclature: Oat, Avena sativa L.; soybean Glycine max L. Merr.; alfalfa, Medicago sativa L.; corn, Zea mays L.