Echolocation call frequency and mitochondrial control region variation in the closely related bat species of the genus Rhinolophus (Chiroptera: Rhinolophidae) occurring in Iran: implications for taxonomy and intraspecific phylogeny

There is a high level of morphological similarity among rhinolophid species leading to problematic taxonomic identifications. We undertook analyses of mitochondrial DNA gene sequences (D-loop), along with morphologic and acoustic examinations in order to evaluate taxonomic status and phylogenetic relationships of the horseshoe bat species in Iran. All analyses based on molecular, morphological, and sonar characteristics revealed five rhinolophid species including Rhinolophus mehelyi, R. euryale, R. ferrumequinum, R. blasii, and R. hipposideros in Iran. Genetic study revealed one lineage in R. mehelyi, two lineages in each of species R. euryale, R. ferrumequinum, and R. blasii, and three lineages in R. hipposideros. Our results showed high haplotype diversity (Hd) in the Iranian rhinolophid species. Compared with other studies of bat D-loop sequences, the genetic mean nucleotide diversity obtained for R. blasii in the current study (pi = 0.0569) was the highest value and for R. euryale was found to be the lowest value (pi = 0.0126) among nucleotide diversity values for other bat species. The average dominant frequency (peak frequency +/- standard deviation) for R. euryale, R. ferrumequinum, R. mehelyi, R. blasii, and R. hipposideros was 106.57 +/- 0.32, 82.02 +/- 0.19, 105.57 +/- 0.45, 92.85 +/- 0.95, and 110.74 +/- 1.93 respectively. As a conclusion, based on the present genetic study, there are one to three lineages including at least one southern and one northern in every rhinolophid species in Iran.