Mitochondrial Genetic Diversity of Bat Species from the Maltese Islands and Applications for their Conservation

Year 2019, Volume: 4 Issue: 3, 276 - 292, 30.10.2019

Clare Marie Mifsud Adriana Vella

https://doi.org/10.28978/nesciences.646348

Cited By: 4

Abstract

This work presents the first genetic species identification and phylogenetic analyses of all six bat species known to inhabit the Maltese archipelago. The results provide a DNA-based reference library of 12S rRNA, 16S rRNA, COI, Cytb and ND1 mitochondrial sequences for Maltese bat species. Phylogenetic analyses revealed that the Maltese bat populations do not harbour cryptic diversity. Analyses of genetic diversity for Maltese bat species showed contrasting matrilineal diversity between species, Hypsugo savii exhibited the highest haplotype diversity (Hd = 0.802), while Rhinolophus hipposideros showed no haplotype diversity and Plecotus gaisleri exhibited low values for haplotype diversity (Hd = 0.091). Comparative phylogeographical analyses of mtDNA gene datasets from this study with sequences of conspecific bat populations outside of Malta indicate that mitochondrial haplotypes of Pipistrellus pipistrellus and Rhinolophus hipposideros are unique to the Maltese Islands. Hypsugo savii, Pipistrellus kuhlii, Myotis punicus and Plecotus gaisleri shared the most common mitochondrial haplotype with surrounding geographical areas, including the Ibero-Maghreb region, the Apennine Peninsula and Sicily. The observed genetic diversity and phylogenetic relationships are discussed in the context of the species’ biology and long-term conservation planning of Maltese bat populations.

Keywords

Mitochondrial DNA, Genetic diversity, DNA barcoding, Bat conservation, Insular populations

References

• Andriollo, T., Naciri, Y., & Ruedi, M. (2015). Two mitochondrial barcodes for one biological species: The case of European Kuhl’s pipistrelles (Chiroptera). PLoS ONE, 10(8), e0134881.
• Arif, I. A., Khan, H. A., Bahkali, A. H., Al Homaidan, A. A., Al Farhan, A. H., Al Sadoon, M., & Shobrak, M. (2011). DNA marker technology for wildlife conservation. Saudi Journal of Biological Sciences, 18(3), 219–225.
• Avise, J. C. (2009). Phylogeography: Retrospect and prospect. Journal of Biogeography, 36(1), 3–15.
• Balakrishnan, R. (2005). Species concepts, species boundaries and species identification: A view from the tropics. Systematic Biology, 54(4), 689–693.
• Benda, P., Kiefer, A., Hanák, V., & Veith, M. (2004). Systematic status of African populations of long-eared bats, genus Plecotus (Mammalia: Chiroptera). Folia Zoologica, 53(1), 1–47.
• Benda, P., Spitzenberger, F., Hanák, V., Andreas, M., Reiter, A., Ševčik, M., Šmid, J., & Uhrin, M. (2014). Bats (Mammalia: Chiroptera) of the Eastern Mediterranean and Middle East. Part 11. On the bat fauna of Libya II. Acta Societatis Zoologicae Bohemicae, 78, 1–162.
• Bogdanowicz, W., Hulva, P., Černá Bolfíková, B., Buś, M. M., Rychlicka, E., Sztencel-Jabłonka, A., Cistrone, L., & Russo, D. (2015). Cryptic diversity of Italian bats and the role of the Apennine refugium in the phylogeography of the western Palaearctic. Zoological Journal of the Linnean Society, 174(3), 635–648.
• Çoraman, E., Furman, A., Karataş, A., & Bilgin, R. (2013). Phylogeographic analysis of Anatolian bats highlights the importance of the region for preserving the Chiropteran mitochondrial genetic diversity in the Western Palaearctic. Conservation Genetics, 14(6), 1205–1216.
• Dietz, C., & von Helversen, S. (2004). Illustrated Identification key to the bats of Europe. Electronic publication Version 1.0., Tuebingen & Erlangen, Germany.
• Edgar, R. C. (2004). MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research, 32(5), 1792–1797.
• Felsenstein, J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Society for the Study of Evolution, 39(1), 1–15.
• García-Mudarra, J. L., Ibáñez, C., & Juste, J. (2009). The Straits of Gibraltar: Barrier or bridge to Ibero-Moroccan bat diversity? Biological Journal of the Linnean Society, 96(2), 434–450.
• Hulva, P., Fornůsková, A., Chudárková, A., Evin, A., Allegrini, B., Benda, P., & Bryja, J. (2010). Mechanisms of radiation in a bat group from the genus Pipistrellus inferred by phylogeography, demography and population genetics. Molecular Ecology, 19(24), 5417–5431.
• Hutterer, R., Ivanova, T., Meyer-Cords, C., & Rodrigues, L. (2005). Bat migrations in Europe - A review of banding data and literature (Vol. 28). Federal Agency for Nature Conservation, Bonn, Germany.
• Ibáñez, C., García-Mudarra, J. L., Ruedi, M., Stadelmann, B., & Juste, J. (2006). The Iberian contribution to cryptic diversity in European bats. Acta Chiropterologica, 8(2), 277–297.
• Juste, J., Ibáñez, C., Muñoz, J., Trujillo, D., Benda, P., Karataş, A., & Ruedi, M. (2004). Mitochondrial phylogeography of the long-eared bats (Plecotus) in the Mediterranean Palaearctic and Atlantic Islands. Molecular Phylogenetics and Evolution, 31(3), 1114–1126.
• Juste, J., Benda, P., Garcia-Mudarra, J. L., & Ibáñez, C. (2013). Phylogeny and systematics of Old World serotine bats (genus Eptesicus, Vespertilionidae, Chiroptera): An integrative approach. Zoologica Scripta, 42(5), 441–457.
• Juste, J., Ruedi, M., Puechmaille, S. J., Salicini, I., & Ibáñez, C. (2019). Two new cryptic bat species within the Myotis nattereri species complex (Vespertilionidae, Chiroptera) from the Western Palaearctic. Acta Chiropterologica, 20(2), 285–300.
• Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Cheung, M., Sturrock, S., Buxton, S., Cooper, A., Markowitz, S., Duran, C., Thierer, T., Ashton, B., Meintjes, P., & Drummond, A. (2012). Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics, 28(12), 1647–1649.
• Kiefer, A., Mayer, F., Kosuch, J., Von Helversen, O., & Veith, M. (2002). Conflicting molecular phylogenies of European long-eared bats (Plecotus) can be explained by cryptic diversity. Molecular Phylogenetics and Evolution, 25(3), 557–566.
• Kiefer, A., & Veith, M. (2001). A new species of long-eared bat from Europe (Chiroptera : Vespertilionidae). Myotis, 39, 5–16.
• Kruskop, S. V., Borisenko, A. V., Ivanova, N. V., Lim, B. K., & Eger, J. L. (2012). Genetic diversity of northeastern Palaearctic bats as revealed by DNA barcodes. Acta Chiropterologica, 14(1), 1–14.
• Kumar, S., Stecher, G., & Tamura, K. (2016). MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular Biology and Evolution, 33(7), 1870–1874.
• Lanfear, R., Calcott, B., Ho, S. Y. W., & Guindon, S. (2012). PartitionFinder: Combined selectio of partitioning schemes and substitution models for phylogenetic analyses. Molecular Biology and Evolution, 29(6), 1695–1701
• Lanfear, R., Frandsen, P. B., Wright, A. M., Senfeld, T., & Calcott, B. (2017). Partitionfinder 2: New methods for selecting partitioned models of evolution for molecular and morphological phylogenetic analyses. Molecular Biology and Evolution, 34(3), 772–773.
• Mayer, Frieder, Dietz, C., & Kiefer, A. (2007). Molecular species identification boosts bat diversity. Frontiers in Zoology, 4(4), 1–5.
• Mifsud, C. M., & Vella, A. (2018). Acoustic characterization of bats from Malta: setting a baseline for monitoring and conservation of bat populations. Bioacoustics, 28(5), 427-442.
• Miller, M. A., Pfeiffer, W., & Schwartz, T. (2010). Creating the CIPRES Science Gateway for inference of large phylogenetic trees. Proceedings of the Gateway Computing Environments workshop (GCE) 2010, New Orleans, 1–8.
• Mitchell-Jones, A. J., & McLeish, A. P. (Eds.). (2004). Bat Workers’ Manual. 3rd Edition, Joint Nature Conservation Committee, Peterborough.
• Mucedda, M., Kiefer, A., Pidinchedda, E., & Veith, M. (2002). A new species of long-eared bat (Chiroptera, Vespertilionidae) from Sardinia (Italy). Acta Chiropterologica, 4(2), 121–135.
• Nei, M., & Kumar, S. (2000). Molecular evolution and phylogenetics. Oxford University Press, New York.
• Paradis, E. (2010). pegas: an R package for population genetics with an integrated-modular approach. Bioinformatics, 26(3), 419–420.
• Puechmaille, S. J., Allegrini, B., Benda, P., Gürün, K., Šrámek, J., Ibañez, C., Juste, J., & Bilgin, R. (2014). A new species of the Miniopterus schreibersii species complex (Chiroptera: Miniopteridae) from the Maghreb region, North Africa. Zootaxa, 3794(1), 108–124.
• Puechmaille, S. J., Gouilh, M. A., Piyapan, P., Yokubol, M., Mie, K. M., Bates, P. J., Satasook, C., New, T., Bu, S. S. H., MacKie, I. J., Petit, E. J., & Teeling, E. C. (2011). The evolution of sensory divergence in the context of limited gene flow in the bumblebee bat. Nature Communications, 2(573), doi:10.1038/ncomms1582.
• Robertson, J. M., Langin, K. M., Sillett, T. S., Morrison, S. A., Ghalambor, C. K., & Funk, W. C. (2014). Identifying evolutionarily significant units and prioritizing populations for management on islands. Monographs of the Western North American Naturalist, 7(1), 397–411.
• Rozas, J., Ferrer-Mata, A., Sánchez-DelBarrio, J. C., Guirao-Rico, S., Librado, P., Ramos-Onsins, S. E., & Sánchez-Gracia, A. (2017). DnaSP 6: DNA Sequence Polymorphism Analysis of Large Data Sets. Molecular Biology and Evolution, 34(12), 3299–3302.
• Saitou, N., & Nei, M. (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4(4), 406–425.
• Salicini, I., Ibáñez, C., & Juste, J. (2011). Multilocus phylogeny and species delimitation within the Natterer’s bat species complex in the Western Palearctic. Molecular Phylogenetics and Evolution, 61(3), 888–898.
• Spitzenberger, F., Strelkov, P. P., Winkler, H., & Haring, E. (2006). A preliminary revision of the genus Plecotus (Chiroptera, Vespertilionidae) based on genetic and morphological results. Zoologica Scripta, 35(3), 187–230.
• Swofford, D. L. (2002). Phylogenetic Analysis Using Parsimony (*and Other Methods). Version 4. Sinauer Associates, Suderland, Massachusetts.
• Turmelle, A. S., Kunz, T. H., & Sorenson, M. D. (2011). A tale of two genomes: Contrasting patterns of phylogeographic structure in a widely distributed bat. Molecular Ecology, 20(2), 357–375.
• Veith, M, Mucedda, M., Kiefer, A., & Pidinchedda, E. (2011). On the presence of pipistrelle bats (Pipistrellus and Hypsugo; Chiroptera: Vespertilionidae) in Sardinia. Acta Chiropterologica, 13(1), 89–99.
• von Helversen, O., Heller, K. G., Mayer, F., Nemeth, A., Volleth, M., & Gombkötö, P. (2001). Cryptic mammalian species: A new species of whiskered bat (Myotis alcathoe n. sp.) in Europe. Naturwissenschaften, 88(5), 217–223.
• Worthington Wilmer, J., & Barratt, E. (1996). A non-lethal method of tissue sampling for genetic studies of chiropterans. Bat Research News, 37, 1–3.