A Preliminary Assessment of the Phylogenetic Relationship of the Bats Referred to Miniopterusfuliginosus (Miniopteridae) in Sri Lanka.

Abstract

The family Miniopteridae is represented by at least 38 species of bats distributed mostly over of the Eastern Hemisphere. In South Asia, this genus is represented by three species, including Miniopterus pusillus, Miniopterus magnater, and Miniopterus fuliginosus. Miniopterus fuliginosus, with type locality in Nepal, was long considered a subspecies of Miniopterus schreibersii but recent molecular phylogenetic studies have recognized the widespread Asian form as a distinct species. The phylogenetic relationship of the Sri Lankan bat referred to M. fuliginosus has never been assessed using molecular techniques and here we have evaluated it for the first time. Tissue samples of Miniopterus from two different locations in Sri Lanka were collected and the 16S rRNA region (540 bp) of the mitochondrial genome was amplified and sequenced to assess the phylogenetic affinities of the Sri Lankan populations. DNA extraction, quantification, PCR and gel electrophoresis were conducted at the Molecular Biology laboratory in Rajarata University of Sri Lanka, and sequencing of PCR products was done at Genetech, Sri Lanka. DNA sequence results obtained from the two samples were processed using the program Geneious Pro. 16S rRNA reference sequences and (9) of seven Miniopterus species from Papua New Guinea (2), Vietnam (1), China (3), Kenya (2) and Tunisia (1) were downloaded from Genbank for comparison and Molossus sinaloae (Molossidae) was used as an out- group as this was considered as the sister group of the family Miniopteridae. Phylogenetic analysis and pairwise genetic distances were calculated using MEGA X software. The Maximum Likelihood tree recovered the two Sri Lankan bats in a strongly supported (bootstrap support >70) monophyletic clade, which was sister to M. fuliginosus (all from China). The two sister clades were separated by a genetic distance of 2.4-2.6%. According to the range of genetic distances between other species of Miniopterus (2.4-7.7%), populations in Sri Lanka referred to M. fuliginosus might actually represent a different species. However, additional geographic and genetic sampling and flanking morphometric analysis will be required to ascertain their relationships.