陕西秦岭非法贸易动物(兽类)基因条码构建及分子鉴定

现今秦岭珍稀野生动物非法贸易频繁发生,野生动物仅通过外表难以准确鉴定。通过构建陕西省秦岭地区羚牛、林麝、斑羚、鬣羚、金丝猴、黑熊、小麂、毛冠鹿、果子狸、豹猫、野猪、大熊猫、黄鼬、鼬獾、獾15种兽类线粒体DNA Cytb基因条码,对3例来自秦岭森林公安收缴的无法鉴定的动物肌肉样品进行分子物种鉴定。利用Neighbor-joining法和非加权配对算数平方法,构建分子系统发生树,对比三种待鉴定样品样本和15种动物的Cytb序列的遗传距离和序列相似性。经分析,待鉴定样品A1与斑羚聚为一枝,序列相似性为99.4%遗传距离为0.006;待鉴定样品A2与鬣羚聚为一枝,序列相似性为98.8%遗传距离为0.012;待鉴定样品A3与黑熊聚为一枝,序列相似性最高为100%遗传距离为0.000。从而鉴定出三种物种分别为斑羚、鬣羚和黑熊。基因条码为物种鉴定和野生动物保护提供便利条件。     

利用红外相机技术分析秦岭有蹄类动物活动节律的季节性差异

2009年8月至2013年4月期间,在陕西观音山自然保护区,利用18台红外相机收集到羚牛(Budorcas taxicolor)、川西斑羚(Naemorhedus griseus)、中华鬣羚(Capricornis milneedwardsii)、毛冠鹿(Elaphodus cephalophus)、小麂(Muntiacus reevesi)、林麝(Moschus berezovskii)6种有蹄类动物的照片数据,通过相对丰富度指数分析了它们的活动规律及季节性差异。结果表明:(1)6种有蹄类动物在研究区域总丰富度达到了58.71%,其中羚牛的相对丰富度是28.02%,川西斑羚13.24%,毛冠鹿10.08%,中华鬣羚4.21%,小麂2.26%,林麝0.90%。(2)6种有蹄类动物的月相对丰富度反映了其年活动格局,其中羚牛、川西斑羚、毛冠鹿、中华鬣羚、小麂表现出一致性,即夏季活动最为频繁,秋季减弱,冬季达到活动低谷,春季逐渐回升;而林麝则在冬季活动最为频繁,夏季最弱。(3)日时间段相对丰富度反映了动物全年的日活动规律,其中川西斑羚和羚牛相似,主要以白天活动为主;毛冠鹿、小麂、林麝具有明显的晨昏活动习性;中华鬣羚活动高峰出现在02:00–06:00和20:00–22:00,以夜间活动为主。(4)分析不同季节6种有蹄类动物日活动规律,羚牛在春季出现一定的差异,活动高峰出现在16:00–20:00;川西斑羚、毛冠鹿、中华鬣羚在冬季表现出一定的差异,活动高峰相对延迟或者提前;小麂春季表现出差异,活动主要集中在00:00–10:00和18:00–20:00;林麝由于数据相对较少,在4个季节表现出不同的活动规律。(5)夜行性分析得到中华鬣羚具有较强的夜间活动能力,夜间相对丰富度达到了65.81%。这些研究结果有助于监测有蹄类动物种群的变化,为保护区有效保护管理提供了数据支持。     

基于28S, COI和Cytb基因序列的薜荔和爱玉子传粉小蜂分子遗传关系研究

薜荔和爱玉子均属于桑科榕属植物,二者为同一物种的原变种与变种的关系,早期研究认为这两种榕树与同一种传粉榕小蜂(Wiebesia pumilae (Hill))建立了稳定的互利共生关系,但近期在形态学、生态学、传粉生物学等方面对二者的研究结果表明,薜荔传粉小蜂和爱玉子传粉小蜂之间可能发生了遗传分化。实验用核糖体28SrDNAD1-D3区、线粒体Cytb及COI基因部分序列,对采自福建3个不同样地的薜荔传粉小蜂和3个不同品系的栽培爱玉子的传粉小蜂进行分析,结果表明:(1)薜荔传粉小蜂和爱玉子传粉小蜂的核糖体28S序列的碱基组成中A,T,G,C 4种含量较平均,C+G的平均含量(56%)稍高于A+T的含量(44%)。线粒体Cytb序列中A+T的含量(76.1%)明显高于C+G的含量(23.9%),COI序列中A+T的含量(71.9%)也明显高于G+C的含量(28.1%),这是膜翅目昆虫线粒体基因的普遍特征。在薜荔和爱玉子传粉小蜂的线粒体Cytb及COI基因中,密码子第三位点A+T的含量最高。(2)比较薜荔和爱玉子传粉小蜂的3种分子标记的变异范围显示,28S进化速度较Cytb及COI序列慢,比较保守,更适合科、亚科等较高分类单元的研究。薜荔传粉小蜂与爱玉子传粉小蜂之间的亲缘关系较近,采用Cytb与COI序列进行分析更为精确。(3)用Cytb及COI序列对薜荔传粉小蜂与爱玉子传粉小蜂之间的遗传距离进行分析显示,薜荔传粉榕小蜂个体间Cytb序列平均遗传距离为0.0054,爱玉子传粉小蜂个体间的Cytb遗传距离为0.0164;薜荔传粉小蜂与爱玉子传粉小蜂群体之间的Cytb序列平均遗传距离为0.1385;COI序列的薜荔传粉榕小蜂个体间遗传距离为0.0048,爱玉子传粉小蜂各样本间平均遗传距离为0.0102;薜荔传粉小蜂与爱玉子传粉小蜂群体间COI序列平均遗传距离为0.1896,两群体间的遗传距离(差异大于10%以上)明显大于群体内各样本之间的遗传距离,表明薜荔传粉小蜂与爱玉子传粉小蜂之间已经发生了很大的遗传分化,其变异水平达到了种间分化水平,即薜荔传粉小蜂与爱玉子传粉小蜂为两个不同的种。     

基于Cytb和COⅠ基因的猎隼分子鉴定

通过与从GenBank中下载的8种隼属鸟类的Cytb和COⅠ部分序列进行同源比对,并以最大简约法(MP)和贝叶斯推断法(BI)构建系统树,对1只涉案的隼科鸟类进行了分子鉴定。结果表明:样品与猎隼Cytb和COⅠ的序列同源性最高,分别为98.91%～99.41%和99.67%～100%,样品与猎隼Cytb和COⅠ序列的遗传距离最小,分别为0.003～0.007和0.000～0.003;同时,样品和猎隼在两种系统树上都始终聚在一起。由此可推断该样品为猎隼,为2010年5月四川省丹巴县森林公安局受理的非法盗猎和贩运隼形目鸟类一案的审理提供了有力的证据。     

高榕榕果内Eupristina属两种榕小蜂的遗传进化关系

对生长在福州地区的高榕进行长期追踪观察,发现高榕榕果内仅生活着Eupristina altissima和Eupristina sp.榕小蜂,前者为高榕的传粉小蜂,后者无传粉行为,两者雌蜂之间在体色、触角、花粉袋和花粉刷等部位存在细微的差异,而两者雄蜂之间无形态差异。通过克隆福建地区5个样地的高榕榕果内收集到的E. altissima和Eupristina sp.榕小蜂,以及细叶榕的传粉小蜂Eupristina verticillata(外群)的Cytb及COI基因,并进行碱基组成及遗传距离分析,用邻接法构建系统发育树,分析两榕小蜂群体之间的遗传进化关系,结果显示:(1)榕小蜂COI及Cytb序列碱基组成中A+T的含量(Cytb序列中A+T=75.3%,COI序列中A+T=75.5%)显著高于G+C,符合膜翅目昆虫线粒体基因碱基组成特征。(2)对两群体小蜂进行遗传距离分析显示,Cytb序列中E. altissima和Eupristina sp. 群体内各样本之间的平均遗传距离分别为0.0092和0.0030,而E. altissima与Eupristina sp. 群体间的平均距离为0.1588;COI序列中E. altissima 与Eupristina sp. 群体内各样本之间的平均遗传距离分别为0.0065和0.0205,而二者群体间的平均遗传距离为0.1043,表明两者群体间的遗传距离明显大于各自群体内各样本间的遗传距离。统计GenBank中下载的6个属34种榕小蜂Cytb序列的种间遗传距离为0.0811-0.1723,6个属28种榕小蜂COI序列的种间遗传距离为0.0939-0.1986。由此认为E. altissima和Eupristina sp.之间的遗传距离差异已经达到了种间水平,即E. altissima与Eupristina sp.为两个不同的种。(3)在形态上,两种小蜂的雌蜂之间有微小差异,而二者雄蜂之间无差异,但Cytb与COI序列分析结果一致表明:E. altissima与Eupristina sp.雄蜂之间,以及二者雌蜂之间的遗传距离均差异显著,表明形态变异滞后于基因变异。雌蜂在表型上进化快于雄蜂,可能是由于雌蜂羽化后从榕果出飞,受到外界环境因素的影响较大,且两种雌蜂在传粉功能上存在差异,故二者之间的形态差异较大,而雄蜂寿命短,又终生生活在黑暗封闭、环境变化相对恒定的榕果内,两种雄蜂在行为上不存在差异,故二者表型变异较为缓慢。E. altissima和Eupristina sp.小蜂对宿主的专一性不强,在榕-蜂协同进化过程中,可能发生过宿主转移事件。     

基于细胞色素b基因的中国岩羊不同地理种群遗传差异分析

为了揭示中国岩羊不同地理种群的遗传差异,探讨岩羊亚种分化的分子机制,采用中国岩羊不同地理种群的细胞色素b(Cyt b)基因的全序列,分析了碱基变异情况、遗传距离以及核苷酸序列差异。用最大似然法和贝叶斯法构建分子系统树并对获得的拓扑结构进行分析。结果发现,西藏亚种与四川亚种Cyt b基因平均序列差异为4.2%(±0.007),处于偶蹄目亚种的序列差异范围内,支持了目前对岩羊西藏亚种的分类地位。四川亚种内部各地理种群之间的遗传距离(0.033±0.0 111)与它们分别到西藏种群的遗传距离(0.042±0.007)差异不显著(t=1.824,P=0.084),说明四川亚种内部各地理种群间已经发生较显著的遗传分化。其中,四川、甘肃和青海种群亲缘关系较近,并与四川亚种内部的其它种群已产生了显著的遗传分化。因此认为四川亚种内部各地理种群的种下分化需要深入研究。     

苔原鼩鼱(Sorex tundrensis)在中国分布的首次证实

2008 年8 月利用陷阱法在内蒙古额尔古纳国家级自然保护区获得了4 号鼩鼱类标本,个体皆小, 尾长小于32 mm,貌似姬鼩鼱,颅全长在16. 50 ～17.24 mm 之间,大于姬鼩鼱(15 mm)。为了确证该4 号标本的物种类型,本文测定了它们的线粒体细胞色素b (Cyt b)基因全序列,结合从GenBank 下载的鼩鼱类序列构建系统进化关系。发现测定序列和下载苔原鼩鼱序列很好地聚在一起并形成单系。标本间及与下载的苔原鼩鼱间遗传距离小。从分子角度确证了标本为苔原鼩鼱,证实了苔原鼩鼱在中国的分布。本文对标本进行了描述。     

基于16S rDNA与Cytb序列的疏广蜡蝉属三个近似种的分子鉴定

本文通过PCR扩增首次获得了疏广蜡蝉属Euricania Melichar,1898 3个近似种:透明疏广蜡蝉E.clara Kato,1932、长刺疏广蜡蝉E.longa Xu,LiangJiang,2006和短刺疏广蜡蝉E.brevicula Xu,LiangJiang,2006的16S r DNA序列和Cytb序列;使用MEGA 6.0软件,分析了其序列组成及变异,计算了遗传距离,以宽广蜡蝉属Pochazia AmyotServille,1843的圆纹宽广蜡蝉P.guttifera Walker,1851和眼斑宽广蜡蝉P.discreta Melichar,1898为外群,利用邻接法(NJ)和最大简约法(MP)构建了系统发育树。结果显示:透明疏广蜡蝉、长刺疏广蜡蝉和短刺疏广蜡蝉的16S r DNA序列长度分别为414 bp,404 bp,435 bp,而Cytb基因序列长度分别为492 bp,468 bp,472 bp,三者的序列比对、碱基组成成分存在差异;2属5种广翅蜡蝉16S r DNA基因的种间遗传距离为0.008-0.098,属间遗传距离为0.122-0.197,而Cytb基因的种间遗传距离为0.038-0.055,属间遗传距离为0.181-0.188,表明16S r DNA和Cytb基因序列可作为DNA条形码候选基因片段,用于广翅蜡蝉近似属、种的分子鉴定。系统发育树的拓扑结构显示,3个近似种的亲缘关系基本一致,各自聚为一支,且置信值均达96%以上。本文亦结合了透明疏广蜡蝉、长刺疏广蜡蝉和短刺疏广蜡蝉形态学特征的差异对其进行了讨论和分析。     

青海高原牦牛mtDNA Cytb基因和D-loop区遗传多样性及系统进化分析

为探究青海高原牦牛的遗传多样性和起源进化关系,本研究通过测定和分析青海高原牦牛155个个体细胞色素b基因(Cytb)和D-loop区全序列,分析多态性及构建系统进化树.结果表明:青海高原牦牛Cytb基因全序列长度为1 140 bp,个体间序列长度无差异,4种核苷酸T、A、G、C的含量分别为26.26％、31.73％、13.09％、28.920％;发现13个SNP位点,全部为转换,符合Cytb基因保守的特征;核苷酸多样性(Pi)为0.002 88,单倍型数(H)为9,单倍型多样性(Hd)为0.645;D-loop区序列长度在892～895 bp之间,不同个体间存在序列长度差异;4种核苷酸T、A、G、C的平均比例分别为28.69％、32.22％、13.75％、25.34％,A+T含量为60.91％,G+C含量为39.09％.在155个个体中共统计出41个SNP位点,其中转换38个,颠换3个,缺失5个,插入3个.其核苷酸多样性(Pi)为0.012 44,分析得到的单倍型数(H)为38,单倍型多样性(Hd)为0.881.在基于Cytb基因的系统进化树中,青海高原牦牛首先与野牦牛和巴州牦牛聚在一起,紧接着与美洲野牛聚在一起;在D-loop区的系统进化分析中,青海高原牦牛首先与西藏牦牛和野牦牛聚在一起,展示出丰富的遗传多样性,同时进一步支持了牦牛和野牦牛划为牛亚科牦牛属(Bovidae)的观点.本研究结果为牦牛改良及育种工作提供了科学依据.     

利用红外相机技术对四川王朗国家级自然保护区野生动物物种多样性的初步调查

本研究以四川王朗国家级自然保护区为研究区域, 利用红外相机对保护区内的主要野生动物进行了初步调查, 分析了该区域的物种多样性现状、相机数量和相机工作日与物种数量间的关系以及物种的相对丰富度。结果表明: 42台红外相机共拍摄到物种独立照片1,793张, 鉴定出野生动物25种, 包括大熊猫(Ailuropoda melanoleuca)、四川羚牛(Budorcas tibetanus)和黄喉雉鹑(Tetraophasis szechenyii) 3种国家一级重点保护野生动物, 黑熊(Ursus thibetanus)、黄喉貂(Martes flavigula)、中华鬣羚(Capricornis milneedwardsii)、中华斑羚(Naemorhedus griseus)等8种国家二级重点保护野生动物。在相机数量增加到23台的时候拍摄到了本次记录的全部25种野生动物, 并且在单台相机工作日达到180天时, 物种数达到饱和。保护区内物种相对丰富度最高的是血雉(Ithaginis cruentus)(29.28)和毛冠鹿(Elaphodus cephalophus)(15.78); 大熊猫的相对丰富度为8.09; 红腹角雉(Tragopan temminckii)、中华鬣羚和黄喉雉鹑的相对丰富度在2-5之间; 中华斑羚、勺鸡(Pucrasia macrolopha)、黑熊、四川羚牛和蓝马鸡(Crossoptilon auritum)的相对丰富度最低, 不到1。综上所述, 红外相机能够有效地对野生动物资源进行监测调查, 对于相对丰富度较低的物种需要投入更多的精力, 这些物种的栖息地保护对于自然保护区的发展至关重要。     

Habitat associations of four ungulates in mountain forests of southwest China, based on camera trapping and dung counts data

The present study aimed to assess abundance indices and habitat associations of four sympatric ungulate species (alpine musk deer Moschus chrysogaster, tufted deer Elaphodus cephalophus, Chinese serow Capricornis milneedwardsii, and Chinese goral Naemorhedus griseus) in Baima Xueshan Nature Reserve of southwest China, using camera trapping and dung counts data. Camera traps were set along six dung transects in different habitats and explored habitat use of the sympatric ungulates using trapping rates. The results obtained revealed that Chinese serow showed a negative association with open canopy cover and low canopy cover. Alpine musk deer were associated with oak shrubs, oak forests and open canopy cover, while tufted deer avoided oak shrubs. Goral showed no significant associations with habitat variables. Alpine musk deer and tufted deer had considerable habitat overlap with Chinese serow. By finding a high correlation between indices, the study indicates that camera trapping may represent a valid index of relative abundance, matching results from other studies.     

在线粒体DNA细胞色素b基因序列水平上对鬣羚系统发育的研究

测定了鬣羚在中国的7个地方种群的线粒体DNA细胞色素b基因的419bp的片段序列,并分析了其序列差异,构建了分子系统进化树。结果表明：鬣羚各种群间的序列差异在0－14．31％,鬣羚种群最早在大约5．7百万年有共同的母系祖先,并且在中国最早可能是从青藏高原东南缘的区域开始演化的。随后向周围地区辐射扩散,日本鬣羚的分化时间大约在3．1百万年前,并且可能是从中国大陆迁移过去的。白玉和道孚的鬣羚种群（与其它种群的碱基替换率＞11．21％）,日本鬣养分种群（与其它种群的碱基替换率＞7．63％）,洛扎的鬣羚种群（与其它种群的碱基替换率＞4．05％）的分化估计已接近或达到亚种水平。鬣羚在中国至少存在3个进化显单元：A．白玉和道孚种群;B．洛扎种群;C．隆子,陕西,皖南和德格种群,建议将它们分开管理,避免杂交,以保存遗传分歧较大的类群及其进化潜力。     

Molecular phylogenetic status of the Korean goral and Japanese serow based on partial sequences of the mitochondrial cytochrome b gene

To investigate the molecular phylogenetic status of the Korean goral, Nemorhaedus caudatus raddeanus, and Japanese serow, Capricornis crispus, we determined partial sequences of the mitochondrial cytochrome b gene of twelve Korean gorals and sixteen Japanese serows, and compared them with those of the major lineages of Rupicaprini species including two other Nemorhaedus species and two other Capricornis species. The Korean gorals examined possessed two haplotypes with only one nucleotide difference between them, while the Japanese serows showed slightly higher sequence diversity with five haplotypes. Genetic distances and molecular phylogenetic trees indicated that there is considerable genetic divergence between the Korean goral and N. caudatus (the Chinese goral) [Groves and Shields (1996)], but virtually none between Korean and Russian gorals. The Korean and Russian gorals may therefore be distinct from the Chinese goral. The data highlight the importance of conservation of the goral populations of these regions, and the need to reconsider the taxonomic status of Korean and Russian gorals. Our study also clearly demonstrated sufficient genetic distance between serows and gorals to justify their assignment to separate genera. Of the three species of Capricornis, the Formosan serow, C. swinhoei is more closely related to C. sumatraensis than to the Japanese serow, suggesting that the Formosan serow is a distinct species. Preliminary data on intraspecific genetic variation in the Japanese serow are also presented.     

Phylogenomics of several deer species revealed by comparative chromosome painting with Chinese muntjac paints

A set of Chinese muntjac (Muntiacus reevesi) chromosome-specific paints has been hybridized onto the metaphases of sika deer (Cervus nippon, CNI, 2n = 66), red deer (Cervus elaphus, CEL, 2n = 62) and tufted deer (Elaphodus cephalophus, ECE, 2n = 47). Thirty-three homologous autosomal segments were detected in genomes of sika deer and red deer, while 31 autosomal homologous segments were delineated in genome of tufted deer. The Chinese muntjac chromosome X probe painted to the whole X chromosome, and the chromosome Y probe gave signals on the Y chromosome as well as distal region of the X chromosome of each species. Our results confirmed that exclusive Robertsonian translocations have contributed to the karyotypic evolution of sika deer and red deer. In addition to Robertsonian translocation, tandem fusions have played a more important role in the karyotypic evolution of tufted deer. Different types of chromosomal rearrangements have led to great differences in the genome organization between cervinae and muntiacinae species. Our analysis testified that six chromosomal fissions in the proposed 2n = 58 ancestral pecoran karyotype led to the formation of 2n = 70 ancestral cervid karyotype and the deer karyotypes is more derived compare with those of bovid species. Combining previous cytogenetic and molecular systematic studies, we analyzed the genome phylogeny for 11 cervid species.     

利用红外相机对湖北后河自然保护区兽类和鸟类的初步调查

湖北后河国家级自然保护区,地处华中腹地,属武陵山东段余脉的一部分山地。该区域生物多样性高,但近15年来未对该地区的自然资源进行过全面调查。为深入了解该区域野生动物多样性现状及其变化,于2012年7月29日至2014年1月8日,在后河自然保护区设置红外相机监测位点53个,对保护区内的鸟类和兽类进行调查。红外相机共拍到鸟兽的独立有效照片1 963张。经鉴定,可以辨认的兽类共计4目12科19种;鸟类共计4目7科29种。在兽类中,野猪（Sus scrofa）,毛冠鹿（Elaphodus cephalophus）,中华斑羚（Naemorhedus griseus）的相对丰富度指数较高;在鸟类中,红腹锦鸡（Chrysolophus pictus）,红腹角雉（Tragopan temminckii）的相对丰富度指数较高。其中,黑熊（Ursus thibetanus）,猕猴（Macaca mulatta）,中华鬣羚（Capricornis milneedwardsii）,中华斑羚,红腹锦鸡,红腹角雉,松雀鹰（Accipiter virgatus）等为国家Ⅱ级重点保护动物;红腿长吻松鼠（Dremomys pyrrhomerus）,岩松鼠（Sciurotamias davidianus）,小麂（Muntiacus reevesi）,宝兴歌鸫（Turdus mupinensis）,棕噪鹛（Garrulax poecilorhynchus）,橙翅噪鹛（Garrulax elliotii）等为中国特有种。本文为今后后河自然保护区野生动物组成、种群动态、活动节律、生境利用、保育等长期监测工作以及后续的科学研究建立基础。     

马麝诸种群地理分化初步探讨

马麝是我国的一种重要经济动物, 是古北界特有种, 主要分布在我国青藏高原及其临近地区。为保护物种多样性, 本文利用分子生物学技术在分子水平探讨了马麝各地理种群之间遗传多样性问题。用线粒体细胞色素b(Cytochrome b) 的DNA 序列进行系统进化分析显示: 马麝四川种群、甘肃种群和西藏两种群在大约100 万年前具有共同祖先。四川种群最先分化出来, 甘肃马麝与西藏马麝的关系最近, 四川马麝与西藏马麝、甘肃马麝间的关系较远, 甘肃马麝与四川马麝约在80 万年前分化, 而甘肃与西藏察隅和类乌齐两种群马麝间分化时间则分别为34万年和57 万年前, 四川马麝与西藏察隅和类乌齐两种群马麝分化时间在90 万年和100 万年前。结果初步表明分布于甘肃、四川和西藏的马麝种群可能为不同的亚种。     

New phylogenetic perspectives on the Cervidae (Artiodactyla) are provided by the mitochondrial cytochrome b gene.

The entire mitochondrial cytochrome b (cyt b) gene was compared for 11 species of the artiodactyl family Cervidae, representing all living subfamilies, i.e., the antlered Cervinae (Cervus elaphus, C. nippon, Dama dama), Muntiacinae (Muntiacus reevesi), and Odocoileinae (Odocoileus hemionus, Mazama sp., Capreolus capreolus, C. pygargus, Rangifer tarandus, Alces alces); and the antlerless Hydropotinae (Hydropotes inermis). Phylogenetic analyses using Tragulidae, Antilocapridae, Giraffidae and Bovidae as outgroups provide evidence for three multifurcating principal clades within the monophyletic family Cervidae. First, Cervinae and Muntiacus are joined in a moderately-to-strongly supported clade of Eurasian species. Second, Old World Odocoileinae (Capreolus and Hydropotes) associate with the Holarctic Alces. Third, New World Odocoileinae (Mazama and Odocoileus) cluster with the Holarctic Rangifer. The combination of mitochondrial cyt b and nuclear k-casein sequences increases the robustness of these three clades. The Odocoileini + Rangiferini clade is unambiguously supported by a unique derived cranial feature, the expansion of the vomer which divides the choana. Contrasting with current taxonomy, Hydropotes is not the sister group of all the antlered deers, but it is nested within the Odocoileinae. Therefore, Hydropotes lost the antlers secondarily. Thus, the mitochondrial cyt b phylogeny splits Cervidae according to plesiometacarpal (Cervinae + Muntiacinae) versus telemetacarpal (Odocoileinae + Hydropotinae) conditions, and suggests paraphyly of antlered deer.     

Complete mitochondrial genome of the leaf muntjac (Muntiacus putaoensis) and phylogenetics of the genus Muntiacus

The leaf muntjac (Muntiacus putaoensis) is an endemic deer species found in the east transHimalayan region.In recent years,population numbers have decreased due to heavy hunting and habitat loss,and little genetic data exists for this species,thus our knowledge of distribution rangs and population sizes likewise remain limited.We obtained mtDNA genes and the complete mitochondrial genome sequence of M.putaoensis using PCR,followed by direct sequencing.The complete mitogenome sequence was determined as a circular 16 349 bp mitochondrial genome,containing 13 protein-coding genes,two rRNA genes,22 tRNA genes,and one control region,the gene composition and order of which were similar to most other vertebrates so far reported.Most mitochondrial genes,except for ND6 and eight tRNAs,were encoded on the heavy strand.The overall base composition of the heavy strand was 33.1％ A,29.3％ T,24.2％ C,and 13.4％ G,with a strong AT bias of 62.4％.There were seven regions of gene overlap totaling 95 bp and 11 intergenic spacer regions totaling 74 bp.Phylogenetic analyses (ML and BI) among the Muntiacus genus based on the sequenced of mitogenome and ND4L-ND4 supported M.putaoensis as a member of Muntiacus,most closely related to M.vuquangensis.However,when analyses based on cyt b included two more muntjacs,M.truongsonensis was most closely related to M.putaoensis rather than M.vuquangensis,and together with M.rooseveltorum,likely forming a M.rooseveltorum complex of the species.This study will help in the exploration of the evolutionary history and taxonomic status of the leaf muntjac,as well as its protection as a genetic resource.     

Molecular phylogenetic analysis of circumnuclear hemoproteids (Haemosporida: Haemoproteidae) of sylviid birds, with a description of Haemoproteus parabelopolskyi sp. nov

Haemoproteus spp., with circumnuclear gametocytes and tentatively belonging to Haemoproteus belopolskyi, are widespread and prevalent in warblers belonging to the Sylviidae, with numerous mitochondrial cytochrome b (cyt b) lineages detected among them.We sampled the hemoproteids from 6 species of warblers adjacent to the Baltic Sea. Parasites were identified to species based on morphology of their gametocytes, and a segment of the parasite's cyt b gene was sequenced. Sixteen mitochondrial cyt b lineages of hemoproteids with circumnuclear gametocytes were recorded. Two clades of lineages (clade A in species of Acrocephalus and Hippolais and clade B in species of Sylvia) with sequence divergence between their lineages >5% are distinguished in the phylogenetic tree. Within the clades A and B, the genetic distance between the lineages is < or = 3.9 and < and = 2.8%, respectively. We compared the morphology of gametocytes of 3 lineages (hHIICT1, hMW1, and hSYAT2) in detail. The lineages hHIICTI and hMW1 (clade A) belong to the morphospecies H. belopolskyi. Parasites of the lineage hSYAT2 (clade B) are described as a new species Haemoproteus parabelopolskyi, which can be readily distinguished from H. belopolskyi by the significantly smaller nuclei of its macrogametocytes. Lineages closely related to H. belopolskyi and H. parabelopolskyi are identified. The sequence divergence between lineages of these 2 morphospecies ranges between 5.3 and 8.1%. It seems probable that avian Haemoproteus spp. with a genetic differentiation of > or =5% in mitochondrial cyt b gene might be morphologically differentiated at the stage of gametocytes. This study establishes the value of both PCR and morphology in identification of avian hemoproteids.