Zoological Journal of the Linnean Society, 2019, 187, 518–534. With 6 figures. Evolutionary history of field mice (Murinae: Apodemus), Downloaded from https://academic.oup.com/zoolinnean/article-abstract/187/2/518/5530313/ by Institute of Zoology, CAS user on 04 October 2019 with emphasis on morphological variation among species in China and description of a new species DEYAN GE1, , ANDERSON FEIJÓ1, , JILONG CHENG1, , LIANG LU2, RONGRONG LIU2, ALEXEI V. ABRAMOV3,4, LIN XIA1, ZHIXIN WEN1, WEIYONG ZHANG5, LEI SHI5 and QISEN YANG1* 1Key Laboratory of Zoological Systematics and Evolution, Institute of Zoology, Chinese Academy of Sciences, Beichen West Road, Chaoyang District, Beijing 100101, China 2State Key Laboratory for Infectious Diseases Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 102206, China 3Zoological Institute, Russian Academy of Sciences, Saint Petersburg 199034, Russia 4Joint Russian–Vietnamese Tropical Research and Technological Centre, Hanoi, Vietnam 5Fanjingshan National Nature Reserve, Tongren, 554400, China Received 22 January 2019; accepted for publication 25 March 2019 Mice of the genus Apodemus are widely distributed across Eurasia. Several species of this genus are hosts of important zoonotic diseases and parasites. The evolutionary history and dispersal routes of these mice remain unclear and the distribution of these species in China was poorly explored in previous studies. We here investigate the divergence times and historical geographical evolution of Apodemus and study the taxonomy of species in China by integrating molecular and morphological data. The crown age of this genus is dated to the Late Miocene, approximately 9.84 Mya. Western and Central Asia were inferred as the most likely ancestral area of this genus. Moreover, we recognize nine living species of Apodemus in China: Apodemus uralensis, A. agrarius, A. chevrieri, A. latronum, A. peninsulae, A. draco, A. ilex, A. semotus and A. nigrus sp. nov., the last from the highlands (elevation > 1984 m) of Fanjing Mountain in Guizhou Province and Jinfo Mountain in Chongqing Province. This new species diverged from A. draco, A. semotus and A. ilex approximately 4.53 Mya. The discovery of A. nigrus highlights the importance of high mountains as refugia and ‘isolated ecological islands’ for temperate species in south-eastern China. KEYWORDS: divergence times – hantavirus – Ljungan virus – molecular clock – molecular phylogeny – mountains – new species – pathogen vectors – refugia. INTRODUCTION A. sylvaticus (Linnaeus, 1758) in northern Africa (Libois et al., 2001) and A. agrarius (Pallas, 1771) Apodemus Kaup, 1829 is widely distributed across on the Danish islands Lolland and Faster (Andersen Eurasia and extends into small areas of the et al., 2017), which is not typical for mice of the genus northernmost part of Africa (Musser et al., 1996; Apodemus. More than 100 species or subspecies of this Musser & Carleton, 2005). The recent distributional genus have been established in the historical literature range expansion of several species in this genus is a (Thomas, 1922; Allen, 1938; Ellerman & Morrison- result of anthropogenic introductions. For example, Scott, 1951; Xia, 1984; Musser et al., 1996; Musser & Carleton, 2005). Species of Apodemus are most *Corresponding author. Email: [email protected] abundant in the broadleaf forests of Palaearctic and [Version of Record, published online 9 July 2019; Oriental areas. Several of these species host multiple http://zoobank.org/urn:lsid:zoobank.org:pub:0122DEEF- human pathogens (Klein et al., 2015; Ma et al., 2015), 3F68-4D2F-A119-378D8C4CA5CF] notably hantavirus (Guzzetta et al., 2017; Tian et al., © 2019 The Linnean Society of London, Zoological Journal of the Linnean Society, 2019, 187, 518–534 518 EVOLUTIONARY HISTORY OF FIELD MICE 519 2017) and Ljungan virus (Hauffe et al., 2010). A study variability of field mice was found in the Mediterranean on the evolutionary history and dispersal of this peninsulas than in northern Europe (Michaux et al., genus is, therefore, important for public health and 2003). Moreover, a large number of fossil species from biodiversity conservation in Eurasia and North Africa. Europe were described that were absent from glacial Rapid radiation of Apodemus during the Late assemblages, but are always present in interglacial Miocene is thought to have been associated with global assemblages (Martin Suarez & Mein, 1998; Knitlova Downloaded from https://academic.oup.com/zoolinnean/article-abstract/187/2/518/5530313/ by Institute of Zoology, CAS user on 04 October 2019 forest changes, when the flora changed from tropical & Horacek, 2017a, b). However, it is unclear how the to temperate during the Late Miocene (Serizawa et al., evolutionary history of Apodemus formed its current 2000). The earliest fossil records of this genus and its distributional pattern. sister genera were dated to the early Late Miocene, Faunas in many regions of China remain relatively approximately 11 Mya (Freudenthal, 1976; Martin poorly explored, impeding a comprehensive Suarez & Mein, 1998; Kimura et al., 2017), but based understanding of interspecific differentiation. on molecular data, the earliest divergence time of this Allen (1938) recognized five species of Apodemus genus was estimated to be approximately 6–8 Mya in China and Xia (1984) listed six: A. sylvaticus, A. (Michaux et al., 2002; Liu et al., 2004; Suzuki et al., 2008; draco, A. peninsulae, A. latronum, A. chevrieri and Darvish et al., 2015). Three crucial evolutionary events A. agrarius. Zheng (1993) reported fossil occurrences of this genus were recognized in previous studies: (1) of A. chevrieri, A. agrarius, A. sylvaticus and A. cf. initial broad dispersal and radiation approximately peninsulae that were dated to the Late Pleistocene in 6 Mya, (2) regional radiation in Europe and China Chongqing and Guizhou Provinces. Based on a large approximately 2 Mya and (3) westward dispersal number of Apodemus specimens from China, Musser of A. agrarius to Europe in the Quaternary (Suzuki et al. (1996) recognized seven species, A. agrarius, et al., 2008). These conclusions were proposed without A. chevrieri, A. peninsulae, A. latronum, A. draco, A. evidence from fossil information or robust statistical semotus and A. uralensis, but excluded A. sylvaticus. analyses. A comprehensive study including extant Smith & Xie (2008) included one more species, A. and fossil species is important for understanding the pallipes (Barrett-Hamilton, 1900), than the study of evolutionary history of this genus. Musser et al. (1996), but this number was not followed According to the most comprehensive mammalian by Wilson et al. (2016), detailed information are given checklists, the composition of Apodemus is still in Table 1. In recent studies, A. ilex is also considered disputable. Mammal species of the world and the a distinct species (Liu et al., 2004, 2012, 2017). Handbook of the mammals of the world listed 20 A previous study based on a wider range of sampling, species of Apodemus (Musser & Carleton, 2005; Wilson and using cytochrome oxidase subunit I (Cox1) et al., 2016). Traditional taxonomy divided Apodemus recognized A. agrarius, A. chevrieri, A. peninsulae, into three groups: the Apodemus group, the Sylvaemus A. latronum, A. ilex, A. draco and A. uralensis, and group and the Alsomys group (Zimmermann, 1962) identified a distinct genetic lineage from Guizhou or the Argenteus group (Musser et al., 1996). Studies Province (Liu et al., 2017). However, it is unclear how based on complete mitochondrial cytochrome b (Cytb) to identify these species based on morphology and a sorted Apodemus species into four groups or subgenera: detailed description of the distinct genetic lineage the Sylvaemus group [A. uralensis (Pallas, 1811), is lacking. An integrative study on the phylogeny of A. flavicolllis (Melchior, 1834), A. alpicola Heinrich, Apodemus and taxonomy of species in this genus in 1952, A. sylvaticus, A. mystacinus (Danford & Alston, China is, therefore, warranted. 1877), A. hermonensis (Filippucci et al., 1989)], the In recent years, we collected a large number Apodemus group [(A. agrarius, A. chevrieri (Milne- of samples of the genus Apodemus in China and Edwards, 1868), A. speciosus (Temminck, 1844), A. examined collections in several museums that draco (Barrett-Hamilton, 1900), A. ilex Thomas, 1922, preserve Chinese specimens. Here, our aim is to (1) A. semotus Thomas, 1908, A. latronum Thomas, 1911 investigate the evolutionary history of this taxon by and A. peninsulae (Thomas, 1907)] and A. argenteus integrating fossil occurrences and extant species, (2) (Temminck, 1844) and A. gurkha Thomas, 1924, use the broadest geographic coverage of Apodemus constitute two other distinct groups (Serizawa et al., to date, to assess the phylogenetic relationships and 2000; Liu et al., 2004; Suzuki et al., 2008). Generally, morphological variation of all species of this genus different aspects of species in Europe and eastern Asia recorded in China and (3) test whether the newly have been well studied. For example, genetic variation discovered genetic lineage in our previous study among species using protein electrophoresis suggested represents a distinct species. By integrating molecular a recent separation of members of the subgenus and morphological data, we established a new species Sylvaemus from a common ancestor, followed by rapid of the genus Apodemus from