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Geomorphologists have long recognized that eolian sand transport pathways extend over long distances in desert regions. Along such pathways, sediment transport by wind can surmount topographic obstacles and cross major drainages. Recent studies have suggested that three distinct eolian sand transport pathways exist (or once existed) in the Mojave and Sonoran Desert regions of the southwestern United States. One hypothesized pathway is eolian sand transport from the eastern Mojave Desert of California into western Arizona, near Parker, and would require sand movement across what must have been at least a seasonally dry Colorado River valley. We tested this hypothesis by mineralogical, geochemical and magnetic analyses of eolian sands on both sides of the Colorado River, as well as sediment from the river itself. Results indicate that dunes on opposite sides of the Colorado River are mineralogically distinct: eastern California dunes are feldspar-rich whereas western Arizona dunes are quartz-rich, derived from quartz-rich Colorado River sediments. Because of historic vegetation changes, little new sediment from the Colorado River is presently available to supply the Parker dunes. Based on this study and previous work, the Colorado River is now known to be the source of sand for at least three of the major dune fields of the Sonoran Desert of western Arizona and northern Mexico. On the other hand, locally derived alluvium appears to be a more important source of dune fields in the Mojave Desert of California. Although many geomorphologists have stressed the importance of large fluvial systems in the origin of desert dune fields, few empirical data actually exist to support this theory. The results presented here demonstrate that a major river system in the southwestern United States is a barrier to the migration of some dune fields, but essential to the origin of others.