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Summary Data are given on the genetic structure, morphology and chorology of the Long-Spurred orchid, Orchis longicornu Poiret (1789) from Sardinia and the occurrence in this island of the morphologically similar Orchis morio L. (1753), often recorded for Sardinia, is investigated. The genetic analysis of 27 enzyme loci in population samples from locations where both species had been recorded showed that: (i) in the population samples from Sardinia the polymorphic loci are in Hardy-Weinberg equilibrium; (ii) these samples are genetically poorly differentiated from each other (average Nei's D = 0.01): less than 2% of the overall genetic variation observed is attributable to differences between populations (G ST = 0.015); (iii) a high rate of gene flow was estimated between Sardinian populations: Nm ≈ 4, possibly owing to seed dispersal by wind; (iv) when Sardinian samples are compared with O. morio from continental Italy, significant differences in genetic variation were observed: average He = 0.16 in the former, 0.12 in the latter; (v) the genetic distance found between Sardinian populations and those of O. morio from continental Italy is relatively high: average Nei's D = 0.18, average Rogers' D = 0.22; (vi) highly significant differences in allele frequencies were found at a number of loci (Mdh-2, Sod-3, Pgm-1, Gpi-1, Gpi-2) between populations from Sardinia and continental Italy, each giving a probability of correct identification that varies from 0.90 to 1 (diagnostic loci). These findings provide evidence that O. morio is not present in the material genetically analyzed from Sardinia, which includes a single species, apparently corresponding to O. longicornu. The slight heterogeneity observed at some loci in the Sardinian population samples can be attributed in some cases (e.g. Mdh-1, Adh, Est-6) to local differences, possibly adaptive; in others (e.g. Pgm-1 and Gpi-1) to genetic drift effects. The hypothesis that some alleles (Gpi-1 100, Dia 101, NADHdh-2 96) found in O. morio and recorded at low frequency in a few Sardinian populations reflect palaeointrogressive phenomena (owing to sporadical immigrant individuals of O. morio having diluted their genes in O. longicornu genome through multiple generations of backcrosses) is not sufficiently supported by the available data. The genetic data are in agreement with the results from morphological studies. These involved the examination of about 1500 specimens (both alive and dried) from all over Sardinia, and their comparison with herbarium specimens of O. longfcornu from patria typica (Algeria) and of O. morio from continental Italy. The typification of O. longicornu is given. The specimens from Sardinia correspond well, on a morphological basis, to the Algerian ones and therefore were all assigned to O. longicornu. Several characters were considered, differentiating O. longicornu from O. morio both in fresh and dried specimens. However, some of them, currently used in diagnostic keys, show much overlap between the two species. This explains the quotations of O. morio from Sardinia, that appear to be misidentifications of O. longicornu. A description of O. longicornu from Sardinia is provided, and its distribution in the island is defined, on the basis of (i) a critical revision of literature records, and (ii) the examination of fresh and dried Sardinian specimens. O. longicornu is shown to be widespread throughout Sardinia, living in many different habitats from 0 to 1500 m a.s.l.