Phylogenetic Relationships Inferred from Ribosomal ITS Sequences and Biogeographic Patterns in Representatives of the Genus Calopteryx (Insecta: Odonata) of the West Mediterranean and Adjacent West European Zone

Western Europe is a reinvasion zone for the riverine dragonfly genus Calopteryx (Insecta: Odonata). Reinvasion may have been from central West Asia or from the West Mediterranean refugium. Phylogenetic relationships of West Mediterranean and West European taxa of the genus Calopteryx from different localities were inferred from sequences of the internal transcribed spacers (ITS1 and ITS2) of the nuclear ribosomal RNA genes. Twenty-six taxa belonging to the species groups C. splendens, C. meridionalis, C. haemorrhoidalis, C. virgo, C. xanthostoma, and C. exul were analyzed, with two North American species, C. amata and C. aequabilis, as outgroup. Sequence data and phylogenetic analyses were used to infer biogeographical patterns. The ribosomal spacers (ITS1 and ITS2) and the intervening 5.8S rDNA gene were amplified by PCR and sequenced. The ITS2 sequences of the West Mediterranean and West European calopterygids show no length variation but the ITS1 region was slightly variable in length. The sequence variation for ITS1 and ITS2 regions between different West Mediterranean and West European calopterygids was 14.5 and 6.1%, respectively. Phylogenetic relationships inferred from ITS sequences only partly confirm morphological data. A monophyletic origin of all West Mediterranean and West European species emerged. They are separated into two main clades; the splendens-like forms and the virgo/meridionalis/haemorrhoidalis group. Intraspecific variability, indicating different stages of speciation, was detected only in West Mediterranean representatives (e.g., C. xanthostoma) but not in invasive representatives in West Europe. The North African endemic C. exul is more closely related to the Italian C. s. caprai than to C. splendens sensu strictu. Based on the present information, Cretan populations are the only splendens-like taxa in addition to C. s. caprai that deserve subspecies status.