Two geographically separated natural populations of Orchis xgennarii, a hybrid between Orchis morio and Orchis papilionacea, were examined to establish the parental lineages using nuclear ribosomal DNA and chloroplast DNA length polymorphisms. Results indicate that O. morio more frequently provides the maternal lineage in the population from the volcano Mount Vesuvius (central Campania, Italy) than in the one from Cilento (southern Campania, Italy); in the latter locality maternal genomes are preferentially provided by O. papilionacea. The possible causes of this difference in reproductive behavior are discussed in the light of the pollination biology of parental species and of environmental influences.