Limitations of Mitochondrial Cytochrome-b Gene in Resolving Phylogenetic Relationships within the Genus Myotis Kaup, 1829

Öz

Despite being one of the most widespread bat genera worldwide, Myotis Kaup, 1829 is a highly problematic group from a systematic perspective. In this study, several genetic characteristics (nucleotide composition and transition/transversion rate) of this genus were determined through mitochondrial Cytochrome-b region analyses and phylogenetic relationships between species were evaluated. The obtained Bayesian Inference tree did not comply with the literature, with pairwise distances between species ranging from 1.4% to 21%. The low genetic distance between Myotis yumanensis and Myotis velifer (1.4%) suggests that these two species must be re-evaluated taxonomically. Some species belonging to the Myotis mystacinus, Myotis nigricans, and Myotis nattereri species complexes were also located in different branches. Furthermore, Myotis martiniquensis sequences did not cluster in the Bayesian Inference tree and low posterior probability values were observed on some branches. These findings indicate that the CYTB gene may have limited resolution when used alone to reconstruct phylogenetic relationships within Myotis. Therefore, the integration of mitochondrial and nuclear markers may provide more robust insights into the phylogeny of this genus.

Anahtar Kelimeler

• Chiroptera
• marker
• phylogeny
• systematics

Myotis Kaup, 1829 Cinsinde Filogenetik İlişkilerin Çözümlenmesinde Mitokondriyal Sitokrom-b Geninin Sınırlamaları

Öz

Myotis Kaup, 1829 Dünya’daki en yaygın yarasa cinslerinden biri olmasının yanısıra, sistematik açıdan oldukça problemli bir gruptur. Bu çalışmada, mitokondriyal Sitokrom-b bölgesi analizleri ile söz konusu cinsin birtakım genetik özelliklerinin (nükleotid bileşimi ve transisyon/transversiyon oranı) ortaya konulmasının yanısıra, türler arası filogenetik ilişkiler değerlendirilmiştir. Elde edilen Bayesian Çıkarım ağacı literatürdeki veriler ile uyuşmamıştır ve türler arası ikili uzaklık değerleri %1.4-21 olarak bulunmuştur. Myotis yumanensis ve Myotis velifer arası düşük genetik uzaklık (%1.4) bu iki türün taksonomik açıdan tekrar değerlendirilmesi gerektiğini ortaya koymaktadır. Myotis mystacinus, Myotis nigricans ve Myotis nattereri tür komplekslerine ait bazı türler de farklı dallara yerleşmişlerdir. Ek olarak, Myotis martiniquensis sekansları Bayesian Çıkarım ağacında bir araya gelmemiştir ve bazı dallarda düşük sonsal olasılık değerleri gözlenmiştir. Bu bulgular, CYTB geninin, Myotis içindeki filogenetik ilişkileri yeniden yapılandırmak için tek başına kullanıldığında sınırlı çözünürlüğe sahip olabileceğini göstermektedir. Bu nedenle, mitokondriyal ve nükleer belirteçlerin entegrasyonu, bu cinsin filogenisi hakkında daha sağlam bilgiler sağlayabilir.

Anahtar Kelimeler

• Chiroptera
• belirteç
• filogeni
• sistematik

Kaynakça

1. Aanen, D.K., Spelbrink, J.N., & Beekman, M. (2014). What cost mitochondria? The maintenance of functional mitochondrial DNA within and across generations. Philosophical Transactions of the Royal Society B: Biological Sciences, 369(1646), 20130438. https://doi.org/10.1098/rstb.2013.0438.
2. Agnarsson, I., Zambrana-Torrelio, C.M., Flores-Saldana, N.P., & May-Collado, L.J. (2011). A time-calibrated species-level phylogeny of bats (Chiroptera, Mammalia). PLoS currents, 3, RRN1212. https://doi.org/10.1371/currents.rrn1212.
3. Baird, A.B., Hillis, D.M., Patton, J.C., & Bickham, J.W. (2008). Evolutionary history of the genus Rhogeessa (Chiroptera: Vespertilionidae) as revealed by mitochondrial DNA sequences. Journal of Mammalogy, 89(3), 744-754. https://doi.org/10.1644/07-mamm-a-135r2.1
4. Baker, R.J., & Bradley, R.D. (2006). Speciation in mammals and the genetic species concept. Journal of mammalogy, 87(4), 643-662. https://doi.org/10.1644/06-mamm-f-038r2.1
5. Ballard, J.W.O., & Whitlock, M.C. (2004). The incomplete natural history of mitochondria. Molecular ecology, 13(4), 729-744. https://doi.org/10.1046/j.1365-294x.2003.02063.x
6. Benda, P., Gazaryan, S., & Vallo, P. (2016). On the distribution and taxonomy of bats of the Myotis mystacinus morphogroup from the Caucasus region (Chiroptera: Vespertilionidae). Turkish Journal of Zoology, 40(6), 842-863. https://doi.org/10.3906/zoo-1505-47
7. Bickham, J.W., Patton, J.C., Schlitter, D.A., Rautenbach, I.L., & Honeycutt, R.L. (2004). Molecular phylogenetics, karyotypic diversity, and partition of the genus Myotis (Chiroptera: Vespertilionidae). Molecular Phylogenetics and evolution, 33(2), 333-338. https://doi.org/10.1016/j.ympev.2004.06.012
8. Boore, J.L. (2006). The complete sequence of the mitochondrial genome of Nautilus macromphalus (Mollusca: Cephalopoda). BMC genomics, 7(1), 1-13. https://doi.org/10.1186/1471-2164-7-182

9. Carrión-Bonilla, C.A., & Cook, J.A. (2020). A new bat species of the genus Myotis with comments on the phylogenetic placement of M. keaysi and M. pilosatibialis. Therya, 11(3), 508-532. https://doi.org/10.12933/therya-20-999
10. Carrión-Bonilla, C.A., Ron, S., & Cook, J.A. (2024). Species richness in Myotis (Chiroptera: Vespertilionidae): species delimitation and expanded geographic sampling reveal high Neotropical diversity. Journal of Mammalogy, 105(2), 241-258. https://doi.org/10.1093/jmammal/gyad124
11. Carstens, B.C., & Dewey, T.A. (2010). Species delimitation using a combined coalescent and information-theoretic approach: an example from North American Myotis bats. Systematic biology, 59(4), 400-414. https://doi.org/10.1093/sysbio/syq024
12. Castellanos, F.X., Simba, L., & Brito, J. (2025). New altitudinal record of Myotis moratellii Novaes et al., 2021 (Chiroptera, Vespertilionidae) in the northwestern foothills of Ecuador. Check List, 21(3), 506-514. https://doi.org/10.15560/21.3.506
13. Chen, X.D., Sun, C.H., Chen, T.Y., Sun, Z.Y., & Lu, C.H. (2025). Complete Mitochondrial Genomes and Phylogenetic Relationships of Myotis siligorensis and Myotis laniger. Ecology and Evolution, 15(9), e72094. https://doi.org/10.1002/ece3.72094
14. Clary, D.O., & Wolstenholme, D.R. (1985). The mitochondrial DNA molecule of Drosophila yakuba: nucleotide sequence, gene organization, and genetic code. Journal of molecular evolution, 22(3), 252-271. https://doi.org/10.1007/bf02099755
15. Demos, T.C., Webala, P.W., Lutz, H.L., Kerbis Peterhans, J.C., Goodman, S.M., Cortés‐Delgado, N., Bartonjo, M., & Patterson, B.D. (2020). Multilocus phylogeny of a cryptic radiation of Afrotropical long‐fingered bats (Chiroptera, Miniopteridae). Zoologica Scripta, 49(1), 1-13. https://doi.org/10.1111/zsc.12388
16. Dowton, M., Castro, L.R., & Austin, A.D. (2002). Mitochondrial gene rearrangements as phylogenetic characters in the invertebrates: the examination of genome'morphology'. Invertebrate Systematics, 16(3), 345-356. https://doi.org/10.1071/is02003
17. Dzeverin, I., & Ghazali, M. (2010). Evolutionary mechanisms affecting the multivariate divergence in some Myotis species (Chiroptera, Vespertilionidae). Evolutionary Biology, 37(2), 100-112. https://doi.org/10.1007/s11692-010-9086-3
18. Evin, A., Baylac, M., Ruedi, M., Mucedda, M., & Pons, J.M. (2008). Taxonomy, skull diversity and evolution in a species complex of Myotis (Chiroptera: Vespertilionidae): a geometric morphometric appraisal. Biological Journal of the Linnean Society, 95(3), 529-538. https://doi.org/10.1111/j.1095-8312.2008.01076.x
19. Fernández-Silva, P., Enriquez, J.A., & Montoya, J. (2003). Replication and transcription of mammalian mitochondrial DNA. Experimental physiology, 88(1), 41-56. https://doi.org/10.1113/eph8802514
20. Gaughan, S.J., Pope, K.L., White, J.A., Lemen, C.A., & Freeman, P.W. (2020). Mitogenome of northern long-eared bat. Mitochondrial DNA Part B, 5(3), 3592-3593. https://doi.org/10.1080/23802359.2020.1830726
21. Gu, X.M., He, S.Y., & Ao, L. (2008). Molecular phylogenetics among three families of bats (Chiroptera: Rhinolophidae, Hipposideridae, and Vespertilionidae) based on partial sequences of the mitochondrial 12S and 16S rRNA genes. Zoological studies, 47(3), 368-378.
22. Gutiérrez, E.G., & Ortega, J. (2025). Mitogenome comparative analysis of 3 Myotis species endemic to Mexico and detecting selection in OXPHOS genes. Journal of Mammalogy, 106(3), 587-602. https://doi.org/10.1093/jmammal/gyae144
23. Győrössy, D., Tu, V.T., Csorba, G., Thapa, S., Estók, P., Földvári, G., & Görföl, T. (2024). The grey zone of taxonomy—The case of the Sikkim Myotis (Chiroptera: Vespertilionidae: Myotis sicarius), first recorded from Southeast Asia. Vertebrate Zoology, 74, 737-749. https://doi.org/10.3897/vz.74.e127269
24. Hamming, R.W. (1950). Error detecting and error correcting codes. The Bell system technical journal, 29(2), 147-160. https://doi.org/10.1002/j.1538-7305.1950.tb00463.x
25. Hasegawa, M., Kishino, H., & Yano, T.A. (1985). Dating of the human-ape splitting by a molecular clock of mitochondrial DNA. Journal of molecular evolution, 22(2), 160-174. https://doi.org/10.1007/bf02101694
26. Hassanin, A., Colombo, R., Gembu, G.C., Merle, M., Tu, V.T., Görföl, T., Akawa, P.M., Csorba, G., Kearney, T., Monadjem, A., & Ing, R.K. (2018). Multilocus phylogeny and species delimitation within the genus Glauconycteris (Chiroptera, Vespertilionidae), with the description of a new bat species from the Tshopo Province of the Democratic Republic of the Congo. Journal of Zoological Systematics and Evolutionary Research, 56(1), 1-22. https://doi.org/10.1111/jzs.12176
27. Haynie, M.L., Tsuchiya, M.T., Ospina-Garcés, S.M., Arroyo-Cabrales, J., Medellín, R.A., Polaco, O.J., & Maldonado, J.E. (2016). Placement of the rediscovered Myotis planiceps (Chiroptera: Vespertilionidae) within the Myotis phylogeny. Journal of Mammalogy, 97(3), 701-712. https://doi.org/10.1093/jmammal/gyv216
28. Hoofer, S.R., & Van Den Bussche, R.A. (2003). Molecular phylogenetics of the chiropteran family Vespertillionidae. Acta Chiropterologica, 5 (Suppl.), 1-63. https://doi.org/10.3161/001.005.s101
29. Huang, T. (n.d.). New record of Myotis longipes (Chiroptera: Vespertilionidae) in Hunan province, China [Unpublished data]. GenBank. Accession No. MH183152.1
30. Huelsenbeck, J.P., & Ronquist, F. (2001). MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics, 17(8), 754-755. https://doi.org/10.1093/bioinformatics/17.8.754
31. Jones, G., Parsons, S., Zhang, S., Stadelmann, B., Benda, P., & Ruedi, M. (2006). Echolocation calls, wing shape, diet and phylogenetic diagnosis of the endemic Chinese bat Myotis pequinius. Acta Chiropterologica, 8(2), 451-463. https://doi.org/10.3161/150811006779398690
32. Kawai, K., Nikaido, M., Harada, M., Matsumura, S., Lin, L.K., Wu, Y., Hasegawa, M., & Okada, N. (2003). The status of the Japanese and East Asian bats of the genus Myotis (Vespertilionidae) based on mitochondrial sequences. Molecular Phylogenetics and Evolution, 28(2), 297-307. https://doi.org/10.1016/s1055-7903(03)00121-0
33. Kazakov, D.V., Artyushin, I.V., Khabilov, T.K., Tadzhibaeva, D.E., & Kruskop, S.V. (2020). Back to life and to taxonomy: new record and reassessment of Myotis bucharensis (Chiroptera: Vespertilionidae). Zootaxa, 4878(1), 129-144. https://doi.org/10.11646/zootaxa.4878.1.5
34. Kazakov, D.V., Kruskop, S.V., Kawai, K., Gorban, A.A., & Gorobeyko, U.V. (2025). Phylogeography, morphometry and the species distribution modelling in Myotis longicaudatus (Chiroptera, Vespertilionidae) in the Eastern Palaearctic. Mammal Research, 70(1), 127-140. https://doi.org/10.1007/s13364-025-00782-5
35. Kipp, E., Milstein, M., Frank, L., Larsen, R., Wolf, T., Faulk, C., Schaffer, C., & Larsen, P. (2025). Going Mobile: Using Portable Genomic Technologies for PCR‐Free In Situ Species Identification and Real‐Time Molecular Systematics. Ecology and Evolution, 15(11), e72442. https://doi.org/10.22541/au.174715081.11227371/v1
36. Korstian, J., Ray, D.A., Lee, T.E. Jr., & Stevens, R.D. (n.d.)a. Myotis dominicensis mitochondrial gene sequence (GenBank accession no. OP554928.1) [Direct submission]. GenBank.
37. Korstian, J., Ray, D.A., Lee, T.E. Jr., & Stevens, R.D. (n.d.)b. Myotis thysanodes mitochondrial gene sequence (GenBank accession no.OP554943.1) [Direct submission]. GenBank.
38. Kovacova, V., Zukal, J., Bandouchova, H., Botvinkin, A.D., Harazim, M., Martínková, N., L. Orlov, O., Piacek, V., Shumkina, A.P., Tiuno, M.P., & Pikula, J. (2018). White-nose syndrome detected in bats over an extensive area of Russia. BMC Veterinary Research, 14(1), 192. https://doi.org/10.1186/s12917-018-1521-1
39. Kumar, S., Stecher, G., Suleski, M., Sanderford, M., Sharma, S., & Tamura, K. (2024). MEGA12: Molecular Evolutionary Genetic Analysis version 12 for adaptive and green computing. Molecular Biology and Evolution, 41(12), msae263. https://doi.org/10.1101/2024.12.10.627672
40. Lack, J.B., Roehrs, Z.P., Stanley Jr, C.E., Ruedi, M., & Van Den Bussche, R.A. (2010). Molecular phylogenetics of Myotis indicate familial-level divergence for the genus Cistugo (Chiroptera). Journal of Mammalogy, 91(4), 976-992. https://doi.org/10.1644/09-mamm-a-192.1
41. Larsen, R.J., Larsen, P.A., Genoways, H.H., Catzeflis, F.M., Geluso, K., Kwiecinski, G.G., Pedersen, S.C., Simal, F., & Baker, R.J. (2012a). Evolutionary history of Caribbean species of Myotis, with evidence of a third Lesser Antillean endemic. Mammalian Biology, 77(2), 124-134. https://doi.org/10.1016/j.mambio.2011.11.003
42. Larsen, R.J., Knapp, M.C., Genoways, H.H., Khan, F.A.A., Larsen, P.A., Wilson, D.E., & Baker, R.J. (2012b). Genetic Diversity of Neotropical Myotis (Chiroptera: Vespertilionidae) with an Emphasis on South American Species. PLoS ONE, 7(10), e46578. https://doi.org/10.1371/journal.pone.0046578
43. Liu, T., Jia, J., Liu, L., Wang, J., Chen, W., Miao, G., Niu Y., Guo W., Zhang K., Sun K., Yu, W., Zhou J., & Feng, J. (2023). New Insights into the Taxonomy of Myotis Bats in China Based on Morphology and Multilocus Phylogeny. Diversity, 15(7), 805. https://doi.org/10.3390/d15070805
44. Lorch, J. (n.d.). Myotis occultus mitochondrial gene sequence (GenBank accession no. OM160888.1) [Direct submission]. GenBank.
45. Masta, S.E., McCall, A., & Longhorn, S.J. (2010). Rare genomic changes and mitochondrial sequences provide independent support for congruent relationships among the sea spiders (Arthropoda, Pycnogonida). Molecular Phylogenetics and Evolution, 57(1), 59-70. https://doi.org/10.1016/j.ympev.2010.06.020
46. Morales, A.E., Jackson, N.D., Dewey, T.A., O’Meara, B.C., & Carstens, B.C. (2017). Speciation with gene flow in North American Myotis bats. Systematic Biology, 66(3), 440-452. https://doi.org/10.1093/sysbio/syw100
47. Morales-Martínez, D.M., López-Arévalo, H.F., & Vargas-Ramírez, M. (2021). Beginning the quest: phylogenetic hypothesis and identification of evolutionary lineages in bats of the genus Micronycteris (Chiroptera, Phyllostomidae). ZooKeys, 1028, 135. https://doi.org/10.3897/zookeys.1028.60955
48. Nabholz, B., Glémin, S., & Galtier, N. (2009). The erratic mitochondrial clock: variations of mutation rate, not population size, affect mtDNA diversity across birds and mammals. BMC evolutionary biology, 9(1), 54. https://doi.org/10.1186/1471-2148-9-54
49. Novaes, R.L.M., Cláudio, V.C., Carrión-Bonilla, C., Abreu, E.F., Wilson, D.E., Maldonado, J.E., & Weksler, M. (2022a). Variation in the Myotis keaysi complex (Chiroptera, Vespertilionidae), with description of a new species from Ecuador. Journal of Mammalogy, 103(3), 540-559. https://doi.org/10.1093/jmammal/gyab139
50. Novaes, R.L.M., Rodriguez-San Pedro, A., Saldarriaga-Cordoba, M.M., Aguilera-Acuna, O., Wilson, D.E., & Moratelli, R. (2022b). Systematic review of Myotis (Chiroptera, Vespertilionidae) from Chile based on molecular, morphological, and bioacoustic data. Zootaxa, 5188(5), 430-452. https://doi.org/10.11646/zootaxa.5188.5.2
51. Novaes, R. L. M., Cláudio, V. C., Bertocchi, N. A., de Oliveira, K., Semedo, T. B. F., Saldanha, J., Wilson, D. E., & Moratelli, R. (2025). Unveiling the shelf life: a new cryptic species of Myotis (Chiroptera, Vespertilionidae) from South America revealed by an integrative taxonomy approach. Journal of Mammalogy, 106(4), 878-897. https://doi.org/10.1093/jmammal/gyaf016
52. Park, S., Noh, P., Choi, Y.S., Joo, S., Jeong, G., & Kim, S.S. (2019). Population genetic structure based on mitochondrial DNA analysis of Ikonnikov’s whiskered bat (Myotis ikonnikovi—Chiroptera: Vespertilionidae) from Korea. Journal of Ecology and Environment, 43(1), 45. https://doi.org/10.1186/s41610-019-0140-5.
53. Parlos, J.A., Nice, C.C., Baccus, J.T., & Forstner, M.R.J. (n.d.). Population genetics of Myotis velifer [Unpublished data]. GenBank. Accession No. EU680299.1
54. Patrick, L.E., & Stevens, R.D. (2014). Investigating sensitivity of phylogenetic community structure metrics using North American desert bats. Journal of Mammalogy, 95(6), 1240-1253. https://doi.org/10.1644/14-mamm-a-007
55. Patterson, B.D., Webala, P.W., Kerbis Peterhans, J.C., Goodman, S.M., Bartonjo, M., & Demos, T.C. (2019). Genetic variation and relationships among Afrotropical species of Myotis (Chiroptera: Vespertilionidae). Journal of Mammalogy, 100(4), 1130-1143. https://doi.org/10.1093/jmammal/gyz087
56. Platt, R.N., Vandewege, M.W., Kern, C., Schmidt, C.J., Hoffmann, F.G., & Ray, D.A. (2014). Large numbers of novel miRNAs originate from DNA transposons and are coincident with a large species radiation in bats. Molecular biology and evolution, 31(6), 1536-1545. https://doi.org/10.1093/molbev/msu112
57. Platt, R.N., Faircloth, B.C., Sullivan, K.A.M., Kieran, T.J., Glenn, T.C., Vandewege, M.W., Lee, Jr. T. E., Baker R.J., Stevens, R.D., & Ray, D.A. (2017). Conflicting Evolutionary Histories of the Mitochondrial and Nuclear Genomes in New World Myotis Bats. Systematic Biology, 67(2), 236-249. https://doi.org/10.1093/sysbio/syx070
58. Posada, D. (2008). jModelTest: Phylogenetic Model Averaging. Molecular Biology and Evolution, 25(7), 1253-1256. https://doi.org/10.1093/molbev/msn083
59. Puechmaille, S.J., Allegrini, B., Boston, E.S.M., Dubourg-Savage, M.J., Evin, A., Knochel, A., Le Bris, Y., Lecoq, V., Lemaire, M., Rist, D., & Teeling, E.C. (2012). Genetic analyses reveal further cryptic lineages within the Myotis nattereri species complex. Mammalian Biology, 77(3), 224-228. https://doi.org/10.1016/j.mambio.2011.11.004
60. Rambaut, A. (2018). Figtree ver 1.4. 4. Institute of evolutionary biology. University of Edinburgh, Edinburgh. Retrieved from https://tree.bio.ed.ac.uk/software/figtree/
61. Rodriguez, R.M., & Ammerman, L.K. (2004). Mitochondrial DNA divergence does not reflect morphological difference between Myotis californicus and Myotis ciliolabrum. Journal of Mammalogy, 85(5), 842-851. https://doi.org/10.1644/1545-1542(2004)085%3C0842:mdddnr%3E2.0.co;2
62. Ruedi, M., & Mayer, F. (2001). Molecular systematics of bats of the genus Myotis (Vespertilionidae) suggests deterministic ecomorphological convergences. Molecular phylogenetics and evolution, 21(3), 436-448. https://doi.org/10.1006/mpev.2001.1017
63. Ruedi, M., Stadelmann, B., Gager, Y., Douzery, E.J.P., Francis, C.M., Lin, L.K., Guillén-Servent, A., & Cibois, A. (2013). Molecular phylogenetic reconstructions identify East Asia as the cradle for the evolution of the cosmopolitan genus Myotis (Mammalia, Chiroptera). Molecular Phylogenetics and Evolution, 69(3), 437-449. https://doi.org/10.1016/j.ympev.2013.08.011
64. Ruedi, M., Csorba, G., Lin, L.K., & Chou, C.H. (2015). Molecular phylogeny and morphological revision of Myotis bats (Chiroptera: Vespertilionidae) from Taiwan and adjacent China. Zootaxa, 3920(1), 301-342. https://doi.org/10.11646/zootaxa.3920.2.6
65. Ruedi, M., Saikia, U., Thabah, A., Görföl, T., Thapa, S., & Csorba, G. (2021). Molecular and morphological revision of small Myotinae from the Himalayas shed new light on the poorly known genus Submyotodon (Chiroptera: Vespertilionidae). Mammalian Biology, 101(4), 465-480. https://doi.org/10.1007/s42991-020-00081-3
66. Ruedi, M., Manzinalli, J., Dietrich, A., & Vinciguerra, L. (2023). Shortcomings of DNA barcodes: a perspective from the mammal fauna of Switzerland. Hystrix, the Italian Journal of Mammalogy, 34(1), 54-61.
67. Sakai, T., Kikkawa, Y., Tsuchiya, K., Harada, M., Kanoe, M., Yoshiyuki, M., & Yonekawa, H. (2003). Molecular phylogeny of Japanese Rhinolophidae based on variations in the complete sequence of the mitochondrial cytochrome b gene. Genes & Genetic Systems, 78(2), 179-189. https://doi.org/10.1266/ggs.78.179
68. Simmons, N.B. (2005). In: Wilson, D.E. & Reeder, D.M. (eds). Mammal species of the world, a taxonomic and geographic reference. Johns Hopkins University Press, Baltimore Maryland, 312-529.
69. Simmons, N.B., & Cirranello, A.L. (2024). Bats of the World, A taxonomic and geographic database version, 1.5. Retrieved from https://batnames.org/
70. Stadelmann, B., Jacobs, D.S., Schoeman, C., & Ruedi, M. (2004a). Phylogeny of African Myotis bats (Chiroptera, Vespertilionidae) inferred from cytochrome b sequences. Acta Chiropterologica, 6(2), 177-192. https://doi.org/10.3161/001.006.0201
71. Stadelmann, B., Herrera, L. G., Arroyo-Cabrales, J., Flores-Martínez, J.J., May, B. P., & Ruedi, M. (2004b). Molecular systematics of the fishing bat Myotis (Pizonyx) vivesi. Journal of Mammalogy, 85(1), 133-139. https://doi.org/10.1644/1545-1542(2004)085<0133:MSOTFB>2.0.CO;2
72. Stadelmann, B., Lin, L.K., Kunz, T.H., & Ruedi, M. (2007). Molecular phylogeny of New World Myotis (Chiroptera, Vespertilionidae) inferred from mitochondrial and nuclear DNA genes. Molecular phylogenetics and evolution, 43(1), 32-48. https://doi.org/10.1016/j.ympev.2006.06.019
73. Tavaré, S. (1986). Some probabilistic and statistical problems in the analysis of DNA sequences. Lectures on Mathematics in the Life Sciences, 17, 57-86.
74. The Mammal Diversity Database of the American Society of Mammalogists (ASM), Version 2.4. (2026). https://doi.org/10.5281/zenodo.17033774
75. Triant, D.A., & DeWoody, J.A. (2006). Accelerated molecular evolution in Microtus (Rodentia) as assessed via complete mitochondrial genome sequences. Genetica, 128(1), 95-108. https://doi.org/10.1007/s10709-005-5538-6
76. Tsytsulina, K., Dick, M.H., Maeda, K., & Masuda, R. (2012). Systematics and phylogeography of the steppe whiskered bat Myotis aurascens Kuzyakin, 1935 (Chiroptera, Vespertilionidae). Russian Journal of Theriology, 11(1), 1-20. https://doi.org/10.15298/rusjtheriol.11.1.01
77. Tu, V.T., Hassanin, A., Görföl, T., Arai, S., Fukui, D., Thanh, H.T., Son, N.T., Furey, N.M., & Csorba, G. (2017). Integrative taxonomy of the Rhinolophus macrotis complex (Chiroptera, Rhinolophidae) in Vietnam and nearby regions. Journal of Zoological Systematics and Evolutionary Research, 55(3), 177-198. https://doi.org/10.1111/jzs.12169
78. Uvizl, M., & Benda, P. (2022). Intraspecific variation of Myotis emarginatus (Chiroptera: Vespertilionidae) inferred from mitochondrial and nuclear genetic markers. Acta Chiropterologica, 23(2), 285-300. https://doi.org/10.3161/15081109acc2021.23.2.002
79. Wang, X., Guo, W., Yu, W., Csorba, G., Motokawa, M., Li, F., Zhang, Q., Zhang, C., Li, Y., & Wu, Y. (2017). First record and phylogenetic position of Myotis indochinensis (Chiroptera, Vespertilionidae) from China. Mammalia, 81(6), 605-609. https://doi.org/10.1515/mammalia-2016-0063
80. Wang, W., Tian, J.H., Chen, X., Hu, R.X., Lin, X.D., Pei, Y.Y., Lv, J.X., Zheng, J.J., Dai, F.H., Song, Z.G., Chen, Y.M., & Zhang, Y.Z. (2022). Coronaviruses in wild animals sampled in and around Wuhan at the beginning of COVID-19 emergence. Virus Evolution, 8(1), veac046. https://doi.org/10.1093/ve/veac046
81. Weyeneth, N., Goodman, S.M., & Ruedi, M. (2011). Do diversification models of Madagascar’s biota explain the population structure of the endemic bat Myotis goudoti (Chiroptera: Vespertilionidae)?. Journal of Biogeography, 38(1), 44-54. https://doi.org/10.1111/j.1365-2699.2010.02393.x
82. Wolstenholme, D.R. (1992). Animal mitochondrial DNA: structure and evolution. International review of cytology, 141, 173-216. https://doi.org/10.1016/s0074-7696(08)62066-5
83. Wu, Z., Han, Y., Wang, Y., Liu, B., Zhao, L., Zhang, J., Su, H., Zhao, W., Liu, L., Bai, S., Dong, J., Sun, L., Zhu, Y., Zhou, S., Song Y., Sui, H., Yang, J., Wang, J., Zhang, S., Qian, Z., Jin, Q. (2022). A comprehensive survey of bat sarbecoviruses across China in relation to the origins of SARS-CoV and SARS-CoV-2. National Science Review, 10(6). https://doi.org/10.1093/nsr/nwac213
84. Velasquez-Restrepo, S., Orozco, M. C., Franco-Sierra, N.D., Martínez-Cerón, J.M., & Díaz-Nieto, J. F. (2024). Identification of non-model mammal species using the MinION DNA sequencer from Oxford Nanopore. PeerJ, 12, e17887. https://doi.org/10.7717/peerj.17887
85. Volleth, M., & Heller, K.G. (2012). Variations on a theme: karyotype comparison in Eurasian Myotis species and implications for phylogeny. Vespertilio, 16, 329-350.
86. Yang, X., Chen, X., Gao, X., Sun, G., Song, X., Dou, H., & Zhang, H. (2023). Presumptive First Record of Myotis aurascens (Chiroptera, Vespertilionidae) from China with a Phylogenetic Analysis. Animals, 13(10), 1629. https://doi.org/10.3390/ani13101629
87. Yakovchuk, P., Protozanova, E., & Frank-Kamenetskii, M.D. (2006). Base-stacking and base-pairing contributions into thermal stability of the DNA double helix. Nucleic acids research, 34(2), 564-574. https://doi.org/10.1093/nar/gkj454
88. Yuzefovich, A.P., Artyushin, I.V., Skopin, A.E., Son, N.T., & Kruskop, S.V. (2022). Taxonomic diversity of the Hipposideros larvatus species complex (Chiroptera: Hipposideridae) in mainland Asia. Zootaxa, 5200(1), 73-95. https://doi.org/10.11646/zootaxa.5200.1.6
89. Zhang, Z., Tan, X., Sun, K., Liu, S., Xu, L., & Feng, J. (2009). Molecular systematics of the Chinese Myotis (Chiroptera, Vespertilionidae) inferred from cytochrome-b sequences. Mammalia: International Journal of the Systematics, Biology & Ecology of Mammals, 73(4). https://doi.org/10.1515/mamm.2009.058
90. Zhukova, S.S., Yuzefovich, A.P., Lebedev, V.S., & Kruskop, S.V. (2025). Reassessment of the Taxonomic Borders Within Pipistrellus (Chiroptera, Vespertilionidae, Pipistrellini). Diversity, 17(5), 317. https://doi.org/10.3390/d17050317