Mitochondrial phylogeny shows multiple independent ecological transitions and northern dispersion despite of Pleistocene glaciations in meadow and steppe vipers (Vipera ursinii and Vipera renardi)
Graphical abstract
Introduction
Pleistocene climatic fluctuations profoundly affected the distributions and structure of most European species (Hewitt, 1999, Taberlet et al., 1998, Nieto, 2011). Traditional biogeographic scenarios suggest that extinction of thermo-sensitive taxa occurred during cold stages of climatic cycles in northern Europe and persisted in southern refugia. Northern areas were covered by an ice sheet and many species could not inhabit the severe periglacial zone. Recolonization of northern areas occurred during warm stages (Ursenbacher et al., 2006b, Joger et al., 2007, Barbanera et al., 2009). Accumulation of data from different taxa have resulted in more complex picture how climatic oscillation and glaciations cycles shaped distribution and genetic diversity across European species (Stewart et al., 2010, Nieto, 2011). For example, cold-tolerant species could persist in refugia located outside of traditionally recognized southern refugia (Taberlet et al., 1998, Hewitt, 1999, Ursenbacher et al., 2006a, Joger et al., 2007, Randi, 2007, Zeisset and Beebee, 2008). Pre-Pleistocene radiations also occurred in many thermophilic European species (Guicking et al., 2006, Joger et al., 2007, Guicking et al., 2009, Recuero et al., 2012).
Changes of entire Eurasian biomes were caused by both shifts of mean temperatures and a complex suite of environmental factors, including precipitation and continentality (duration of seasons, differences in temperature and humidity between summer and winter etc.), among other factors (Velichko and Spasskaya, 2002). Xerophytic grasslands (steppes) and their species are frequently distributed in areas of low mean annual temperatures, but warmer in summer than temperate forests. Their distribution is determined by an equal or slightly negative balance of precipitation and evaporation. Intervals of major glacial cooling were drier globally (Suc et al., 1999, Velichko, 1989, Velichko and Spasskaya, 2002). Therefore, the largest distribution of steppes and steppe-like open landscapes occurred at times of dry glaciations (Velichko, 1989, Artyushenko and Turlo, 1989). Warm interglacial periods, such as now, favoured the expansion of forests and together with sea transgressions in the past could have lead to the shrinkage and fragmentation of steppes, as exemplified by regions near the northern Black Sea and Caspian Sea (Blagovolin et al., 1982, Artyushenko and Turlo, 1989, Velichko and Spasskaya, 2002).
The mitochondrial phylogeny projected on a geographic dimension (genogeography sensu Serebrovsky, 1928; phylogeography sensu Avise et al., 1987) can yield insights into how Pleistocene climatic cycling impacted the distribution and biogeography of the Eurasian steppe biota. Because mitochondrial DNA (mtDNA) is clonally inherited and passed exclusively through matrilines in most animals, historical patterns of dispersal are not obfuscated by genetic recombination and gene sorting, which occurs in the nuclear DNA (nDNA) genome. Consequently, mitochondrial phylogenies can track the response of species to climate changes and yield predictions into future reactions at least from the perspective of female dispersal. Analyses of mtDNA sequence tell only one part of a potentially more complex story (William et al., 2004, Godinho et al., 2008), yet they provide valuable insights into the evolutionary history of species, including consequences of habitat changes, adaptation, impact of climate fluctuations and dispersion. Biome-restricted indicator species serve as ideal models for such investigations.
Vipers of the Vipera renardi complex mostly occupy lowland steppes from central Ukraine eastwards across southern Russia and Kazakhstan to western China. Vipera renardi is the indicator species of snakes of the continuous Eurasian steppe belt. The range of V. r. renardi covers great homogenous territories of steppe almost without disruption (Bannikov et al., 1977, Nilson and Andrén, 2001) and coincides with steppe zone itself. In contrast, European V. ursinii mostly occurs in montane grasslands. Historical scenarios on the origin and colonization in steppe and meadow vipers fall into two categories: (a) periodic expansion and subsequent fragmentation of the northern lowland range during the Pleistocene along with isolation and speciation in southern montane refugia (Nilson and Andrén, 2001, Tuniyev et al., 2010); or (b) having an earlier montane southern origin with a later dispersion to the north (Nilson and Andrén, 2001, Kukushkin, 2009).
Meadow and steppe vipers of the V. ursinii-renardi group are a morphologically and ecologically well defined assemblage. Belonging to genus Vipera Laurenti, 1768, sometimes the group is assigned to subgenus Acridophaga Reuss, 1927 (Nilson and Andrén, 2001) or placed in subgenus Pelias Merrem, 1820. Morphology-based reviews since the mid-20th century have been inconclusive with respect to the recognition of taxa, especially in the poorly explored Asian and Caucasian regions (Kramer, 1961, Saint Girons, 1978; Dely and Stohl, 1989). Although these vipers were also among the first reptiles to be studied taxonomically using molecular data (Joger et al., 1992, Herrmann et al., 1992), a molecular phylogeny remains elusive. Until recently, few representatives were included in phylogenetic (Kalyabina-Hauf et al., 2004, Garrigues et al., 2005) and population genetic studies (Ujvari et al., 2005, Ferchaud et al., 2010). Ferchaud et al. (2012) and Gvozdik et al. (2012) focused on the V. ursinii complex. They evaluated few samples of the V. renardi complex and in doing so they neither covered taxonomic diversity nor geographic range of renardi complex, especially in the Caucasus.
The group subdivides into two widely accepted complexes. The Vipera ursinii complex of meadow vipers consists of Vipera ursinii ursinii (Bonaparte, 1835) and the Balkan taxa Vipera ursinii macrops Mehely, 1911, Vipera ursinii rakosiensis Mehely, 1893, Vipera ursinii graeca Nilson and Andrén, 1988, and Vipera ursinii moldavica Nilson, Andrén et Joger, 1993. The eastern steppe vipers, i.e. the Vipera renardi complex, consists of Vipera renardi (Christoph 1861), with the subspecies V. r. bashkirovi Garanin et al. (2004), V. r. puzanovi Kukushkin (2009), V. r. tienshanica Nilson et Andrén 2001, and V. r. parursinii Nilson et Andrén 2001, as well as Vipera eriwanensis (Reuss 1933), Vipera ebneri Knőpfler et Sochurek 1955, Vipera lotievi Nilson et al. (1995), Vipera altaica Tuniyev et al. (2010) (Nilson and Andrén, 2001, Joger and Dely, 2005, Dely and Joger, 2005, Kalyabina-Hauf et al., 2004). Taxonomy is not without controversy. Whereas Joger and Dely (2005) reduced Vipera lotievi to being a subspecies of V. renardi, some authors continue to recognize the species (Tuniyev et al., 2009, Tuniyev et al., 2011). Vipera anatolica Eiselt et Baran 1970 from southwestern Turkey was considered a subspecies of V. ursinii (Billing, 1985) or a species of the renardi complex together with V. renardi and V. eriwanensis (Nilson and Andrén, 2001). Kalyabina-Hauf et al. (2004) resolved V. anatolica as the sister-group of the ursinii, renardi and kaznakovi complexes based on analyses of mtDNA sequences. Recently described taxa include V. r. bashkirovi (Garanin et al., 2004), V. r. puzanovi (Kukushkin, 2009), the “Altai taxon of V. renardi” of Nilson and Andrén (2001) as V. altaica (Tuniyev et al. (2010)) and morphologically similar to V. eriwanensis population of steppe viper from Şamaxi as Vipera shemakhensis (Kukushkin et al., 2012, Tuniyev et al., 2013). The phylogenetic positions of these new taxa remain unknown.
The primary aim of our work is to reconstruct the mitochondrial phylogeny of ursinii-renardi group, outline distribution of mitochondrial clades and give insight into the dispersal history and historical demography of populations on the background of climatic oscillations of Pleistocene, landscape rearrangements and the transition between highland meadows and lowland steppes.
Section snippets
Sampling and molecular protocols
We used 429 vipers samples, taken from specimens from collections of the Zoological Institute of Russian Academy of Sciences, St. Petersburg, Russia (ZISP); Sochi National Park, Sochi, Russia (SNP), Natural History Museum Gothenburg, Sweden (NMG), Zoological Museum of State Moscow University, Moscow, Russia (ZMMSU), The Museum of Nature at V. N. Karazin Kharkiv National University, Kharkiv, Ukraine (MNKNU), Zoological Museum of National Natural History Museum, Kiev, Ukraine (ZMNHMK); stored in
Phylogeny
The analysis of 420 DNA sequences resolved 104 unique haplotypes for Cytb (66 haplotypes of renardi complex and 38 haplotypes of ursinii complex) and 36 haplotypes of COI (24 haplotypes of renardi complex and 12 haplotypes of ursinii complex). The concatenated alignment of Cytb and COI consisted of 2021 aligned positions among which 957 were invariable and 205 potentially parsimony informative. Selected pairwise uncorrected p-distances between and within taxa are presented in Table 2; clade
The volume of ursinii-renardi group and relationships between major lineages of Pelias
Unlike Ferchaud et al. (2012), we incorporated in our dataset sequences of V. kaznakovi considered as a sister clade to ursinii-renardi group (Joger et al., 2007, Gvozdik et al., 2012, Ferchaud et al., 2012) and V. anatolica, believed to be a representative of the ursinii-renardi group (Nilson and Andrén, 2001, Ferchaud et al., 2012). Our reconstructions placed V. anatolica outside of the entire clade of ursinii-renardi together with all lineages within kaznakovi complex. V. u. graeca and one
Acknowledgments
We are grateful to Valentina Orlova, Yulia Zima, Daniel Melnikov, Konstantin Milto, Evgeny Roitberg, Leif Westrin, Alexander Westerström, Michael Franzen, Konstantin Lotiev, Maria Kolesnikova, Alexey Korshunov, Glib Mazepa, Sergey Ryabov, Serge Utevsky, Aslanbek Magometovich Batkhiev, Artem Kidov, Andrei Baybuz, Halpern Balint, Aram Aghasyan, Vlad Starkov, Lesley Lowcock. We are grateful to Sylvain Ursenbacher and second anonymous reviewers for careful assessment of the manuscript.
We dedicate
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2021, Zoologischer AnzeigerCitation Excerpt :Species tolerant to cold and dry climates can survive outside of the traditionally recognized southern refugia (Taberlet et al. 1998; Hewitt 1999; Ursenbacher et al. 2006a; Joger et al., 1997; Randi 2007; Zeisset & Beebee 2008; Copilaş-Ciocianu et al. 2017b). The existence of modern populations of cold-resistant species is usually explained by re-colonization after the last Ice Age (2.6Mya–11,7Kya) from southern, western, and/or eastern refugia (Zinenko et al. 2015; Jandzik et al. 2018; Parvizi et al., 2019; Jablonski et al. 2019; Levin et al. 2019). However, recent studies show that it was possible for at least some insect and plant species to survive in or near their present-day northern distribution areas (Quinzin et al. 2017; Simonsen & Huemer 2014).
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