Natural hybrids have formed in Pullman, Washington, United States between the recently formed allotetraploids Tragopogon miscellus and T. mirus. In addition to forming spontaneously, these hybrids are semifertile, propagating via achenes. Previous work indicated that the tetraploid hybrids have genetic contributions from three progenitor diploids: T. dubius, T. pratensis, and T. porrifolius. Because the hybrids contain genomes from three species, they should be karyotypically variable and have very low fertility. To better understand how these hybrids are semifertile, we applied fluorescent probes to determine chromosome composition.