Molecular phylogeny of the subfamily Schizothoracinae (Teleostei: Cypriniformes: Cyprinidae) inferred from complete mitochondrial genomes

The schizothoracine fishes, members of the Teleost order Cypriniformes, are one of the most diverse group of cyprinids in the Qinghai–Tibetan Plateau and surrounding regions. However, taxonomy and phylogeny of these species remain unclear. In this study, we determined the complete mitochondrial genome of Schizopygopsis malacanthus. We also used the newly obtained sequence, together with 31 published schizothoracine mitochondrial genomes that represent eight schizothoracine genera and six outgroup taxa to reconstruct the phylogenetic relationships of the subfamily Schizothoracinae by different partitioned maximum likelihood and partitioned Bayesian inference at nucleotide and amino acid levels. The schizothoracine fishes sampled form a strongly supported monophyletic group that is the sister taxon to Barbus barbus. A sister group relationship between the primitive schizothoracine group and the specialized schizothoracine group + the highly specialized schizothoracine group was supported. Moreover, members of the specialized schizothoracine group and the genera Schizothorax, Schizopygopsis, and Gymnocypris were found to be paraphyletic.     

南门峡裂腹鱼亚科鱼类形态相似种的分类学地位——形态趋同进化实例

受高原抬升所致的水系变迁及人类活动的影响,分布于南门峡河流的裂腹鱼亚科鱼类与黄河干支流种群间的基因交流受到长期限制。作为孤立小群体,探讨其分类学地位及其在小生境中的进化机制对了解青藏高原鱼类多样性和物种的形成、进化具有重要意义。本文采用聚合酶链式反应(PCR)和直接测序方法获得了南门峡裂腹鱼亚科鱼类(n=29)及其近缘种(n=19)共48个个体的线粒体DNA(mtDNA)细胞色素b(cytb)基因的全序列(1140bp),并以厚唇裸重鱼和尖裸鲤为外群构建了MP和Bayesian系统进化树。南门峡裂腹鱼亚科鱼类29个个体的序列经排序后,发现有100个(8·77%)多态性位点,共定义了16个单倍型,在系统进化树上分布于截然不同的两个族群中。其中5个单倍型(NMX3、6、7、13、15)与其近缘种花斑裸鲤和青海湖裸鲤形成单系群(MP99%,Bayesian98%),而其余11个单倍型(NMX1、2、4、5、8、9、10、11、12、14、16)与黄河干支流的黄河裸裂尻鱼形成另一个单系群(MP99%,Bayesian99%)。序列差异分析显示,分布于不同族群的南门峡裂腹鱼亚科鱼类之间存在较大的碱基差异(平均为7·42%),显示出种间差异水平,表明分布于南门峡河流的裂腹鱼亚科鱼类可能是花斑裸鲤和黄河裸裂尻鱼形态相似种的复合体。结合青藏高原隆升所致的气候环境变化和高原北部水系变迁的事件,推断形态趋同进化可能导致了南门峡河流裂腹鱼亚科鱼类形态相似种的共存,而小生境自然选择压力是引发适应性形态趋同进化的主要原因。     

青海湖裸鲤和黄河裸裂尻鱼感染多子小瓜虫的病理学比较研究

为探讨不同裂腹鱼类感染多子小瓜虫后的病理学差异, 利用多子小瓜虫对青海湖裸鲤指名亚种(Gymnocypris przewalskii przewalskii)和黄河裸裂尻鱼(Schizopygopsis pylzovi Kessler)实施感染实验, 并对2种鱼进行了深入的病理学研究。研究结果显示: (1) 2种鱼的死亡量均呈现先激增后明显回落的趋势, 青海湖裸鲤死亡高峰在感染后第3至第4天, 黄河裸裂尻鱼死亡高峰在第3至第5天, 青海湖裸鲤的死亡比黄河裸裂尻鱼急剧。(2)感染后2种鱼体表均出现大量肉眼可见的白点。青海湖裸鲤皮肤黏液分泌量明显增加, 体表形成胶状黏液层, 黏液层中见不同细胞期小瓜虫包囊。黄河裸裂尻鱼鳍出现蛀鳍现象, 皮肤出现细菌感染样溃烂。(3)解剖发现, 感染组青海湖裸鲤和黄河裸裂尻鱼肝脏发生病理改变呈淡黄色, 胆有不同程度肿大。(4)组织切片和电镜观察显示, 小瓜虫在鳃部位的寄生导致青海湖裸鲤和黄河裸裂尻鱼鳃丝黏连, 鳃小片和鳃丝上皮细胞萎缩、脱落, 鳃丝结构被严重破坏。小瓜虫在2种鱼皮肤的寄生使寄生部位组织突起, 周围组织塌陷。青海湖裸鲤表皮下层出现空隙, 表皮结构被严重破坏。黄河裸裂尻鱼皮肤表皮细胞出现空泡化病理改变, 失去原有紧密结构, 表皮层和固有层间界限变模糊。综上所述, 小瓜虫的感染对青海湖裸鲤和黄河裸裂尻鱼的鳃和皮肤组织造成严重损伤, 但2种鱼表现的症状和造成的组织损伤类型有明显差异, 这与2种鱼长期适应不同水体环境密切相关。     

黄河裸裂尻鱼细胞色素C氧化酶Ⅰ、Ⅱ和Ⅲ亚基基因的克隆及序列特征分析

采用RT-PCR技术克隆获得了黄河裸裂尻鱼(Schizopygopsis pylzovi)CO Ⅰ、Ⅱ、Ⅲ基因的编码序列,并对此进行了初步分析.结果表明,黄河裸裂尻鱼CO I基因全长为1 551 bp,开放阅读框(ORF)由基因全长组成,编码516个氨基酸;COⅡ基因全长为691 bp,开放阅读框为690 bp,编码2...     

黄河裸裂尻鱼群体遗传结构和Cyt b序列变异

测定了来自黄河上游和柴达木盆地托索湖的裸裂尻鱼共16个个体的Cytb基因全序列(1141bp),探讨了种群结构和遗传多样性。用MEGA2.1软件分析了碱基组成和序列变异;以青海湖裸鲤、花斑裸鲤和极边扁咽齿鱼为外类群,用PAUP*4.0b10程序构建了单倍型NJ树;用Arlequin Ver.2000程序计算了群体间遗传变异值(Fst)和Nm值以及群体分化概率值。结果显示,来自柴达木水系托索湖的裸裂尻鱼没有形成单系群,Fst=0.204(P0.05),Nm=1.95。初步判断,黄河和柴达木水系托索湖的裸裂尻鱼未显著分化,支持将柴达木裸裂尻鱼(Schizopygopsis kessleri)归并入黄河裸裂尻鱼(Schizopygopsis pylzovi)的形态学结果。两种群核苷酸多样度()分别为0.0012和0.0026,表现为较低水平。根据校正的分子钟推测,黄河和托索湖裸裂尻鱼群体分歧时间为距今7万年左右的更新世末期,结合地理分布的资料和古地质事件,对黄河裸裂尻鱼群体分布水系间的历史联系进行了分析。       

Mitochondrial DNA phylogeny of two morphologically enigmatic fishes in the subfamily Schizothoracinae (Teleostei: Cyprinidae) in the Qinghai-Tibetan Plateau

The complete mitochondrial DNA (mtDNA) cytochrome b gene (1140 bp) was sequenced in Herzenstein macrocephalus and Gymnocypris namensis and in 13 other species and sub-species ( n = 22), representing four closely related genera in the subfamily Schizothoracinae. Conflicting taxonomies of H. macrocephalus and G. namensis have been proposed because of the character instability among individuals. Parsimony, maximum likelihood and Bayesian methods produced phylogenetic trees with the same topology and resolved several distinctive clades. Previous taxonomic treatments, which variously placed these two species of separate genera or as sub-species, are inconsistent with the mtDNA phylogeny. Both H. macrocephalus and G. namensis appear in a well-supported clade, which also includes nine species of Schizopygopsis , and hence should be transferred to the genus Schizopygopsis . Morphological changes are further illustrated, and their adaptive evolution in response to the local habitat shifts during the speciation process appears to be responsible for conflicting views on the systematics of these two species and hence the contrasting taxonomic treatments. These species are endemic to the Qinghai-Tibetan Plateau, a region with a history of geological activity and a rich diversity of habitats that may have result in the parallel and reversal evolution of some morphological characters used in their taxonomies. Our results further suggest that speciation and morphological evolution of fishes in this region may be more complex than those previously expected.     

The complete mitochondrial genomes of two schizothoracine fishes (Teleostei,Cypriniformes): A novel minisatellite in fish mitochondrial genomes

Minisatellites, a class of variable number tandem repeats (VNTRs), are abundant throughout the control region in animal mitochondrial DNA (mtDNA) but rare in other regions of animal mtDNA. Here, we reported a novel minisatellite in fish mitochondrial genomes. We first determined the complete mitochondrial genomes of two schizothoracine fishes (Herzensteinia microcephalus and Schizopygopsis pylzovi) and found a type of minisatellites in a novel region between the tRNA‐Thr and tRNA‐Pro genes in their mtDNA. To explore the origin and evolution of the minisatellites in different schizothoracine and closely related fishes, we analyzed the available 80 fish mitogenomes which represent five closely related tribes of cyprinine fishes. The results from the phylogenetic analyses show that the schizothoracine fishes sensu stricto is not a monophyletic group and is divided into two clades (Schizothoracini and Schizopygopsini); and the minisatellite is only present in Schizopygopsini distributed in the region between the two tRNA genes (tRNA‐Thr and tRNA‐Pro) of the mtDNA. This is the first record of a minisatellite in a non‐control region of fish mitogenome.     

黄河裸裂尻鱼肥胖基因克隆及其对青藏高原季节性冷环境的适应性表达

目的克隆黄河裸裂尻鱼肥胖基因(obese gene,ob),分析编码蛋白leptin的序列特征,检测ob mR-NA的组织特异性表达和不同环境温度下肝胰脏和白肌ob mRNA的相对表达水平,为探究黄河裸裂尻鱼在青藏高原季节性寒冷环境下的生态适应机制提供科学数据。方法利用RT-PCT、5’-RACE和3’-RACE法获得ob基因全长cDNA序列。通过已知cDNA序列设计半定量RT-PCR和real-time PCR引物,进行表达研究。结果黄河裸裂尻鱼ob基因序列全长为1 337 bp,其中开放阅读框(ORF)长513 bp,编码170个氨基酸。黄河裸裂尻鱼ob mRNA在心脏、肾脏、脂肪、肠、精巢、肝胰脏、鳃、脑和肌肉9个被检组织中均有表达,其中在心脏中表达最强,肌肉组织中表达最弱。栖息水域环境温度下降,将显著抑制黄河裸裂尻鱼肝胰脏和白肌ob mRNA的表达。结论黄河裸裂尻鱼ob基因特殊的组织表达特征可能与其特定组织的能量代谢及其特殊的生物学功能有关。Leptin在黄河裸裂尻鱼能量代谢调节和季节性环境适应方面发挥着重要作用。     

Convergent, parallel and correlated evolution of trophic morphologies in the subfamily schizothoracinae from the Qinghai-Tibetan plateau

Schizothoracine fishes distributed in the water system of the Qinghai-Tibetan plateau (QTP) and adjacent areas are characterized by being highly adaptive to the cold and hypoxic environment of the plateau, as well as by a high degree of diversity in trophic morphology due to resource polymorphisms. Although convergent and parallel evolution are prevalent in the organisms of the QTP, it remains unknown whether similar evolutionary patterns have occurred in the schizothoracine fishes. Here, we constructed for the first time a tentative molecular phylogeny of the schizothoracine fishes based on the complete sequences of the cytochrome b gene. We employed this molecular phylogenetic framework to examine the evolution of trophic morphologies. We used Pagel's maximum likelihood method to estimate the evolutionary associations of trophic morphologies and food resource use. Our results showed that the molecular and published morphological phylogenies of Schizothoracinae are partially incongruent with respect to some intergeneric relationships. The phylogenetic results revealed that four character states of five trophic morphologies and of food resource use evolved at least twice during the diversification of the subfamily. State transitions are the result of evolutionary patterns including either convergence or parallelism or both. Furthermore, our analyses indicate that some characters of trophic morphologies in the Schizothoracinae have undergone correlated evolution, which are somewhat correlated with different food resource uses. Collectively, our results reveal new examples of convergent and parallel evolution in the organisms of the QTP. The adaptation to different trophic niches through the modification of trophic morphologies and feeding behaviour as found in the schizothoracine fishes may account for the formation and maintenance of the high degree of diversity and radiations in fish communities endemic to QTP.     

黄河裸裂尻鱼五种群mtDNA控制区的遗传结构

黄河裸裂尻鱼（Schizopygopsis pylzovi）主要分布于青藏高原东北部兰州以上黄河干支流水系和高原北部柴达木内流水系,由于其对高原特殊生境表现出独特的适应性而受研究者关注。本文采用聚合酶链式反应（PCR）和直接测序的方法获得了一个分布于柴达木内流水系种群和四个黄河支流种群共99条个体的线粒体DNA（mtDNA）控制区（D-100p）821bp核苷酸的序列,探讨了其种群遗传结构和分化。99条个体的序列经比对后,发现77个（9．37％）多态性位点,共定义了53个单倍型,其中有一个单倍型（DT12）为两个种群（大通种群和互助种群）所共享,其余52个单倍型均为某个种群所特有。遗传多样性分析表明,柴达木种群的单倍型多样度（h=0．79±0．06）和核苷酸多样度（π=0．0027±0．0017）均略低于黄河支流各种群。AMOVA分析结果显示,遗传差异主要发生在种群之内,占75．39％,而小同水系的种群间只有24．61％,各种群间成对的Fst及遗传距离均表明种群间出现了一定程度的种群分化,但系统进化树最示出一种混杂的单倍型分布格局,提示青藏高原东北部地质事件所造成的水系间的隔离形成较晚。单倍型歧点分布呈现为单峰以及中性检验Tajima的D（-1．497,P=0．058）和Fu的Fs（-24．741,P=0．001）结果综合表明,黄河裸裂尻鱼在青藏高原隆升过程中曾经历过近期的种群扩张事件[动物学报54（6）：972—980,2008]。     

利用Cyt b基因序列探讨柴达木裸裂尻鱼的分类学地位

长期以来,柴达木裸裂尻鱼(Schizopygopsis kessleri)的分类学地位存在争议。为此,测定了柴达木裸裂尻鱼格尔木河及托素湖种群24个个体及黄河裸裂尻鱼(S.pylzovi)黄河及大通北川河种群12个个体的Cytb基因全序列。通过遗传距离和系统发育分析,探讨了柴达木裸裂尻鱼的分类学地位。结果显示,柴达木裸裂尻鱼与黄河裸裂尻鱼在系统进化树上并未形成相应的单系群;柴达木裸裂尻鱼与黄河裸裂尻鱼种间平均遗传距离为0.54%,明显低于裂腹鱼亚科其他属鱼类的种间遗传距离及本属其他种间遗传距离,表明来自柴达木盆地格尔木河和托素湖的裸裂尻鱼与黄河裸裂尻鱼之间没有达到种级水平的显著分化。综合地质学资料和柴达木水系与黄河水系现今的隔离格局,建议将柴达木裸裂尻鱼作为黄河裸裂尻鱼的一个亚种,即S.pylzovi kessleri。     

Molecular and morphological analysis of endangered species <Emphasis Type="Italic">Gymnodiptychus integrigymnatus</Emphasis> (Teleostei: Cyprinidae)

Gymnodiptychus integrigymnatus is a critically endangered species endemic to the Gaoligongshan Mountains. It was thought to be only distributed in several headwater-streams of the Longchuanjiang River (west slope of the Gaoligongshan Mountains, belonging to the Irrawaddy River drainage). In recent years, dozens of G. integrigymnatus specimens have been collected in some streams on the east slope of the Gaoligongshan Mountains (the Salween drainage). We performed a morphological and genetic analyses (based on cytochrome b and D-loop) of the newly discovered populations of G. integrigymnatus to determine whether the degree of separation of these populations warrants species status. Our analysis from the cytochrome b gene revealed that nine individuals from the Irrawaddy drainage area and seven individuals from the Salween drainage area each have only one unique haplotype. The genetic distance between the two haplotypes is 1.97%. Our phylogenetic analysis revealed that G. integrigymnatus is closely related to highly specialized schizothoracine fishes. Analysis from the mitochondrial control region revealed that G. integrigymnatus has relatively high genetic diversity (π was 0.00891 and h was 0.8714), and individuals from different river drainages do not share the same haplotypes. The AMOVA results indicated 87.27% genetic variability between the Salween and Irrawaddy populations. Phylogenetic trees show two major geographic groups corresponding to the river systems. We recommend that G. integrigymnatus should be considered as a high priority for protected species status in the Gaoligongshan Mountains National Nature Reserve, and that the area of the Gaoligongshan Mountains National Nature Reserve should be expanded to cover the entire distribution of G. integrigymnatus. Populations of G. integrigymnatus from different river systems should be treated as evolutionarily significant units.     

黄河上游花斑裸鲤Cyt b基因的序列变异和遗传多样性

花斑裸鲤（Gymncypris eckloni）主要分布于黄河上游高原宽谷河段深水缓流处或静水湖泊中,在高原淡水生态系统的食物链中具有重要的地位。本文获得了黄河上游花斑裸鲤5个种群共68个个体线粒体DNA（mtDNA）细胞色素b基因的全序列（1 140 bp）,分析了序列变异和种群遗传多样性。68个序列经比对后,发现30个（2.63%）多态性位点,共定义了18个单倍型。结果显示,花斑裸鲤种群单倍型多样度和核苷酸多样度均低于其它鲤科鱼类,这可能与青藏高原经所经历的地质变迁和古气候环境的改变有关,由此生活在高原水域中的鱼类或多或少经历过瓶颈效应。AMOVA分析结果显示,遗传差异主要发生在种群之内,而不是来自不同地理组群间或组群内种群间。单倍型网络图和系统发育分析均没有显示出单倍型与地理位置的对应关系,提示黄河上游花斑裸鲤自然种群未出现分化,应作为一个整体进行保护。单倍型歧点分布呈现为单峰以及中性检验Fu’s Fs（&#8722;15.3400, P<0.001）和Tajima’s D（&#8722;0.6254, P =0.3080）结果综合表明,花斑裸鲤可能经历过近期的种群扩张事件。     

用RAPD技术对特化等级裂腹鱼类亲缘关系的探讨

为确定特化等级裂腹鱼类的遗传分化关系 ,对 7种 2 2个裂腹鱼个体进行随机扩增多态DNA(RAPD)研究。从所使用的 40个随机引物中 ,选择了 37个扩增带谱清晰的引物进行分析。根据构建的分子系统树显示 ,特化等级裂腹鱼类划分为 3个属级分类单元较为合适 ,这与采用形态学特征进行系统分析所得出的结论一致。在 2 2个个体之间遗传距离矩阵中 ,最大的遗传距离指数达到了 90 38% ,此外 ,还有较多的遗传距离指数也在 6 0 %～ 90 %之间。通过分析 ,认为用RAPD标记技术来分析属级或属级以上分类单元的亲缘关系 ,具有一定的适用性。对于一些遗传差异较大的类群 (如裂腹鱼类 ) ,应用属级或属级以上分类单元的亲缘关系 ,很可能会出现个体之间遗传距离接近或达到 10 0 %的情况 ,从而无法探讨其相互之间的亲缘关系。     

蜂猴属（Nycticebus）D-loop控制区和Cyt b基因遗传学研究

测定了蜂猴属线粒体DNA中的D-loop控制区部分序列和细胞色素b基因全序列（1140 bp）,对其变异情况进行了分析,并采用Mega 4.0软件中构建了分子系统树。无论是基于D-loop控制区部分序列构建的系统树,还是基于细胞色素b基因序列构建的分子系统树的拓扑结构图,都清晰地表明蜂猴属由两个分支组成,分支置信度较高,一支由N．pygmaeus聚成,另一支由N．coucang聚成,即支持蜂猴属由N．coucang和N．pygmaeus两物种组成。     

2种珍稀裂腹鱼类线粒体DNA部分序列片段的比较分析

对扁吻鱼(Aspiorhynchus laticeps)及塔里木裂腹鱼(Schizothorax biddulphi)线粒体CO I(624 bp)、Cyt b(712 bp)和D-loop(457 bp)基因片段进行了PCR扩增和序列测定.通过分析比较发现,3个片段A+T含量与其他鱼类相比均较高,G含量偏低这一现象在...     

Historical biogeography of the livebearing fish genus Poeciliopsis (Poeciliidae: Cyprinodontiformes)

To assess the historical biogeography of freshwater topminnows in the genus Poeciliopsis, we examined sequence variation in two mitochondrial genes, cytochrome b (1140 bp) and NADH subunit 2 (1047 bp). This widespread fish genus is distributed from Arizona to western Colombia, and nearly half of its 21 named species have distributions that border on the geologically active Trans-Mexican Volcanic Belt (TMVB), a region that defines the uplifted plateau (Mesa Central) of Mexico. We used the parametric bootstrap method to test the hypothesis that a single vicariant event associated with the TMVB was responsible for divergence of taxa found to the north and south of this boundary. Because the single-event hypothesis was rejected as highly unlikely, we hypothesize that at least two geological events were responsible for divergence of these species. The first (8-16 million years ago) separated ancestral populations that were distributed across the present TMVB region. A second event (2.8-6.4 million years ago) was associated with northward dispersal and subsequent vicariance of two independent southern lineages across the TMVB. The geological history of this tectonically and volcanically active region is discussed and systematic implications for the genus are outlined.     

High altitude adaptation of the schizothoracine fishes (Cyprinidae) revealed by the mitochondrial genome analyses

The schizothoracine fishes, also known as “mountain carps” are widely distributed in the Qinghai-Tibetan Plateau and its peripheral regions. Although they provide a prime example of high altitude adaptation, the phylogenetic relationships and the divergence times among these carp lineages are still controversial. Moreover, the genetic basis for high altitude adaptation is also poorly understood. In this study, we determined the mitochondrial genomes from two species of the schizothoracine fishes, representing a “morphologically primitive” clade and “morphologically specialized” clade, respectively. The phylogenetic tree and the divergence times were estimated within the evolutionary framework of the entire order Cypriniformes. Our results indicate a polyphylyetic relationship of the schizothoracine fishes and suggest two independent migration events into the Qinghai-Tibetan Plateau: one by the “morphologically primitive” clade in the Late Miocene and another by the “morphologically specialized” clade in the Eocene. Rapid speciation events of each clade from the Late Miocene to the Pliocene correspond to the timing of the geologic acceleration of the Qinghai-Tibetan Plateau. Interestingly, we found evidence for positive selection acting on the protein coding genes in the mitochondrial genomes of the “morphologically specialized” clade, implying a possible genetic basis for high altitude adaptation in this derived lineage of cypriniform fishes.     

Molecular phylogeny and biogeography of the Cuban genus Girardinus Poey, 1854 and relationships within the tribe Girardinini (Actinopterygii, Poeciliidae)

Phylogenetic relationships among members of the freshwater fish tribe Girardinini were inferred to test existing colonization and diversification hypotheses for this group in the Caribbean. The genetic material examined was mitochondrial (cytochrome b, 1140 bp) and nuclear (RAG-1 and beta-actin, 2450 bp) DNA from 161 specimens representing 44 ingroup and three outgroup taxa. Our mtDNA and combined data matrix (mtDNA+nuclear DNA) results rendered a well-supported phylogeny for the tribe Girardinini and suggest the need to review the group's current taxonomy. From the data presented here, it may be inferred that the Girardinini diverged from other poeciliid fishes approximately 62 Mya ago in the Palaeocene period. This estimate, however, conflicts with the hypothesis that today's vertebrate fauna is the result of the more recent colonization of the Antillean islands during the Early Oligocene (35-33 Mya ago). The isolation of western, central and eastern Cuba during the Miocene and that of the Juventud Island and Guanahacabibes Peninsula during the Pliocene, are the main geologic events that could have promoted speciation in this group.     

Molecular phylogeny and biogeography of woolly flying squirrel (Rodentia: Sciuridae), inferred from mitochondrial cytochrome b gene sequences

To investigate the genetic diversity between the populations of woolly flying squirrels (Eupetaurus) from the eastern and western extremes of the Himalayas, partial mitochondrial cytochrome b gene sequences (390-810 bp) that were determined from the museum specimens were analyzed using maximum parsimony (MP) and maximum likelihood (ML) methods. The molecular data reveal that the two specimens that were collected in northwestern Yunnan (China) are members of the genus Eupetaurus. Reconstructed phylogenetic relationships show that the populations of Eupetaurus in the eastern and western extremes of the Himalayas are two distinct species with significant genetic differences (12%) and diverged about 10.8 million years ago. Eupetaurus is significantly different from Petaurista and Pteromys. The level of estimated pairwise-sequence divergence observed between Eupetaurus and Petaurista or Pteromys is greater than that observed between Eupetaurus and Trogopterus, Belomys, Glaucomys, or Hylopetes. Considering the divergence time of the two Eupetaurus groups, the glaciations and the uplift of the Himalayas and Qinghai-Tibet plateau during the Pliocene-Pleistocene period might be the major factors affecting the present distribution of Eupetaurus along the Himalayas.