Molecular approach to the identification and characterization of natural hybrids between Orchis pauciflora Ten. and Orchis quadripunctata Cyr. ex Ten. (Orchidaceae)

Seven individuals of Orchis x pseudoartatolica Fleishm., a natural hybrid b etween Orchis pauciflora Ten. and O. quadripunctata Cyr. ex Ten. (Orchidace ae), collected in two close localities in Southern Italy, were characterize d by employing molecular methods in order to identify their parents and to define their maternal lineages. Restriction analysis of PCR-amplified fragm ents containing the Internal Transcribed Spacer II (ITS2) of the nuclear ri bosomal DNA showed that the plants are indeed hybrids between the two above mentioned species and allowed exclusion of O. provincialis Balbis, formerl y indicated as potential parent, from the lineage. The chloroplast trnL (UA A) intron pattern of the hybrids indicates that O. pauciflora invariably pr ovides the maternal line.