Crop rotation promotes productivity, nutrient cycling, and effective pestmanagement. However, in row-crop systems, rotation is frequently limited totwo crops. Adding a third crop, especially a perennial crop, might increasecrop-rotation benefits, but concerns about disruption of agricultural andecological processes preclude grower adoption of a three-crop rotation. Theobjective of the present research was to determine whether weed seed banksdiffer between a sod-based rotation (bahiagrass–bahiagrass–peanut–cotton)and a conventional peanut–cotton rotation (peanut–cotton–cotton) and theimportance of crop phase in weed seed-bank dynamics in a long-termexperiment initiated in 1999 in Florida. Extractable (ESB) and germinable(GSB) seed banks were evaluated at the end of each crop phase in 2012 and2013, and total weed seed or seedling number, Shannon-Weiner's diversity (H′), richness, and evenness were determined. ESBincreased in H′ (36%), richness (29%), and total number ofweed seeds (40%) for sod-based compared with conventional rotation, whereasGSB increased 32% in H′, 27% in richness, and 177% in totalnumber of weed seedlings. Crop phase was a determinant factor in thedifferences between crop rotations. The first year of bahiagrass (B1)exhibited increases in weed seed and seedling number, H′,and richness and had the highest values observed in the sod-based rotation.These increases were transient, and in the second year of bahiagrass (B2),weed numbers and H′ decreased and reached levels equivalentto those in the conventional peanut–cotton rotation. The B1 phase increasedthe germinable fraction of the seed bank, compared with the other cropphases, but not the total number of weed seeds as determined by ESB. Theincreases in H′ and richness in bahiagrass phases weremainly due to grass weed species. However, these grass weed species were notassociated with peanut and cotton phases of the sod-based rotation. Theresults of the present study demonstrated that including bahiagrass as athird crop in a peanut–cotton rotation could increase weed communitydiversity, mainly by favoring increases in richness and diversity, but thestructure and characteristics of the rotation would prevent continuousincreases in the weed seed bank that could affect the peanut and cottonphases.