Allozymic variation in seven enzymes coded by eight loci was studied in seven populations of terrestrial Epipactis orchids, including four populations of E. helleborine, one population of E. palustris and two of E. microphylla, from Latium (Central Italy). Heterozygosity estimates reveal that E. helleborine (EH) is the most variable species (HET=0.233; P=0.59; A=1.8), followed by E. palustris (EP) (HET=0.085; P=0.29; A=1.3) while E. microphylla (EM) is monomorphic for all the loci examined. It is suggested that the two EM populations examined, with a total lack of genetic variability, could originate from the same genetically impoverished ancestral population. Also the low variability amount of EP can be ascribed to genetic drift, probably due to a founder event. Nei's identity coefficients were also calculated. These show a high similarity in EH (Ī=0.987) and EM (Ī=1.000) populations. Average identity value between EP and EM is 0.289, between EP and EH is 0.336 and between EM and EH is 0.784. These data reveal that E. palustris presents a remarkable genetic, besides geographical and ecological, differentiation.