Our overall objective was to use DNA Amplification Fingerprinting (DAF) to determine the relationships between Petunia × hybrida and four wild petunia species,P. axillaris, P. inflata, P. parodii, and P. violacea. This research was to optimize the DAF amplification reaction for petunias, check for variability in the fingerprints among different seedlings of the same species and screen primers to be used for Identifying polymorphisms between cultivars of P. × hybrida end the four wild species. Optimization of the DAF procedure was accomplished by varying concentrations of DNA template (O - 10 ng), MgCl2(0 - 10 mM), and primers (0 - 30 μM). Optimum concentrations were found to be 1.0 ng DNA template and 2.0 mM MgCl2. Clearly resolved banding patterns were produced using primer concentrations from 3.0 μM to 30 μM. When separate seedlings of each wild species from the same seed source were fingerprinted, profiles were consistent. Seeds from other sources are presently being collected to investigate variation between sources. Twenty-five heptamer and octomer primers varying in GC content were screened and ten produced clear banding patterns for the Petunia species. These primers have produced polymorphic profiles between the pink-flowering species and the white-flowering species. Several primers have shown distinct polymorphisms between P. axillaris and P. parodii, the two white-flowering species, which have very similar morphological traits. Similarities in the banding patterns have been found between P. × hybrida and these wild species.