大林姬鼠两亚种线粒体DNA 细胞色素b 基因和控制区的序列多样性

为了研究大林姬鼠两亚种(韩国的指名亚种及中国东北和内蒙古地区的东北亚种)线粒体DNA 的变异程度并确定朝鲜亚种的分类地位,我们分别将来自韩国和中国东北长白山地区的两亚种的线粒体DNA 的细胞色素b 基因和控制区进行了测序分析。我们将测序所得到细胞色素b 基因序列与来自基因库的大林姬鼠5 个亚种的相应的单倍型进行了分析,结果显示大林姬鼠可分为4 个类群[类群1:韩国大林姬鼠指名亚种;类群2:中国长白山和内蒙古地区的东北亚种、俄罗斯外加贝尔的majuculus 亚种;类群3:中国长春的东北亚种、俄罗斯Primorye(俄罗斯远东地区) rufulus 亚种、俄罗斯库页岛(俄罗斯远东地区)和日本北海道地区的giliacus 亚种;类群4:中国黑龙江海林地区的东北亚种]。线粒体的控制区序列分析显示韩国指名亚种也不同于中国东北地区的东北亚种。本研究的类群1,2 和3 与Serizawa et al. (2002)的研究的K、S 和R 的分支相对应。这表明韩国指名亚种(类群1 和分支K)的线粒体DNA 与其他类群不同。另外,我们还发现在细胞色素b 基因构建的系统树中,东北亚种可以与类群2 (分支S)及类群3 (分支R 的不同亚种聚合在一起。我们认为线粒体DNA 的母性遗传与两个相邻亚种的个体之间的种内杂交造成了基于细胞色素b 序列对东北亚种的聚类分析结果与基于形态学特征的分类结果的不一致。因此,我们提出对这些显示出核苷酸序列多样性的东北亚种不能只用细胞色素b的数据进行亚种分类,还应该结合形态学和核DNA 特征进行进一步分析。最后,我们还发现韩国的指名亚种的细胞色素b 序列在平均距离16. 93% 的基础上不同于来自基因库的A. speciosus。Jones and Johnson (1965)指出了韩国的大林姬鼠在形态上的区别,所以我们认为韩国的大林姬鼠指名亚种A. p. peninsulae 是一种具有形态和遗传特异性的地方亚种。     

中国和韩国黑线姬鼠两亚种的线粒体DNA限制性片段分析

本文利用印迹杂交技术对中国华北地区黑线姬鼠Ａｐｏｄｅｍｕｓａｇｒａｒｉｕｓｐａｌｉｄｉｏｒ和韩国黑线姬鼠Ａ．ａｇｒａｒｉｕｓｃｏｒｅａｅ共１０７号标本的线粒体ＤＮＡ（ｍｉｔｏｃｈｏｎｄｒｉａｌＤＮＡ,ｍｔＤ－ＮＡ）,通过８种限制性内切酶的消化,进行了限制性片段的分析。共检出３５种限制性片段和１２种单倍体类型。１２种单倍型在平均离散度为１．０１％时聚合为两个亚群：一个亚群为黑线姬鼠华北亚种,由采自中国４个不同地区的５１号标本的４种单倍型所组成;另一个亚群为黑线姬鼠朝鲜亚种,由采自韩国４个不同地区的５６号标本的８种单倍型所组成。黑线姬鼠华北亚种和朝鲜亚种在ｍｔＤＮＡ表型上表现出一定差异,这在分子水平上确立了两亚种的分类地位。为了进一步澄清黑线姬鼠种下分类的混乱,很有必要对中国其他地区的标本进行该项研究     

新疆草兔的种群遗传结构和亚种分化

新疆草兔 (Lepus capensis) 的群体遗传结构至今无系统的研究报道,亚种水平的分类也长期存在争议.该文测定了形态分类上的新疆草兔3个亚种共87个个体的线粒体DNA (mtDNA)控制区(control region,D-Loop)592 bp的序列,经分析发现148个多态性位点,共定义了44个单倍型.新疆草兔的单倍型多样度(h,0.977 ± 0.005)和核苷酸多样度(π,0.064 ± 0.031)都较高,显示了较高的遗传多样性.分子变异分析(AMOVA)结果显示,4个地理群体间的显著分化可能是由地理隔离造成的.群体遗传结构分析显示,新疆草兔包含4个进化枝,并且每个进化枝都对应特定的分布区域,显示了明显的系统地理结构.该研究的结果支持形态分类上草兔西域亚种(L.c.lehmanni)的分类地位; 但中亚亚种(L.c.centrasiaticus)被分为两个独立的进化枝,提示可能存在两个亚种; 帕米尔亚种(L.c.pamirensis)与其他亚种间的遗传距离在13%以上,提示其可能已达到种的分化水平.     

大石鸡亚种分化及一新亚种描述(鸡形目,雉科)

通过地理分布、形态差异、分子进化证明了我国特有种大石鸡Alectoris magna分化为两个亚种,指名亚种A.magna magna和兰州亚种新亚种A.magna lanzhouensis subsp.nov..对新亚种与指名亚种的形态进行了比较.测新亚种12个和指名亚种7个样本的mtDNA控制区486个碱基,两个亚种间无共享单倍型,其间基因交流受到限制;新亚种各取样种群间共享一种单倍型,说明它们来自共同祖先;新亚种序列变异0.27%,而指名亚种为0.91%,其间差异显著(t=1.77,p=0.046＜0.05);两个亚种间的遗传距离为0.0103,约50万年前它们分歧进化.     

韩国与中国东北地区松鼠线粒体DNA控制区的序列变异

为了研究松鼠东北亚种(Sciurus vulgaris manchuricusThomas)不同种群的序列变异水平并进一步确定分类地位,我们分析了韩国5个地点和中国东北2个地点的松鼠标本的线粒体DNA控制区的全序列(1 058 bp)。39个韩国松鼠标本显示出21种单倍型,这些单倍型间的平均Tamura-Nei距离为1·0%; 24个中国松鼠标本显示21种单倍型,单倍型间的平均Tamura-Nei距离为1·4% (1 058 bp的全序列中发生变异的位点有119个,占11·2%)。韩国松鼠和中国松鼠间的平均距离为1·3%。并且韩国和中国松鼠的所有42个单倍型形成了一个单系分支,Fst值为0·04,表明在两个国家的松鼠间没有发生遗传分化。因此,序列分析的分子生物学的结果支持现行的分类,即来自韩国的朝鲜亚种(S·v·coreae)是中国北部地区松鼠东北亚种(S·v·manchuricus)的同物异名。这还需要进一步对北朝鲜和中国东北其它地区更多标本的分子和形态学分析来验证这一结论。     

中国柽柳沙鼠线粒体DNA 的地理变异及其亚种分化

柽柳沙鼠广泛分布于中亚荒漠地区,在我国分布于蒙新干旱区西部。本研究采用线粒体Cyt b 基因和D-loop控制区两个片段共1 569 bp作为分子标记,对我国蒙新区西部45 个柽柳沙鼠的核苷酸多样性、单倍型多样性及系统发生进行分析,并探讨其亚种分化。45 个柽柳沙鼠线粒体DNA 共定义33 个单倍型,包含142 个核苷酸变异位点(占全序列的9.05% ),其中单态位点34 个,简约信息位点108 个,未发现碱基插入或缺失的现象。基于单倍型的系统发生树和网络关系图显示,柽柳沙鼠的谱系分为两大支:伊犁盆地的种群聚为独立的一支,准噶尔盆地和额济纳地区的种群聚为一支,两大支的分歧时间为0.24 Ma年前,大约在庐山冰期。通过Beast 模拟,柽柳沙鼠在近15 000年有一次快速的种群衰退,表明柽柳沙鼠种群受到了末次盛冰期的影响。基于Cyt b 基和D-loop 序列的证据,我们认为分布于新疆伊犁盆地的柽柳沙鼠为哈萨克亚种(M. t. jaxartensis),分布于我国新疆准噶尔盆地以及甘肃、内蒙地区的为敦煌亚种(M. t. satchouensis)。     

中国东部黑线姬鼠的形态学与生化指标的比较研究

本文从中国东部三个地区黑线姬鼠(Apodemus agrarius) 的形态学和生化指标两方面的差异比较探讨了我国东部黑线姬鼠种下分类问题。背部毛色、黑线清晰度和某些外形量度性状差异系数的比较以及运用聚丙烯酰胺双垂直板电泳分析乳酸脱氢酶、超氧化物歧化酶、苹果酸脱氢酶同工酶以及肌浆蛋白、血清蛋白的结果表明, 三地黑线姬鼠种群间存在一定差异。这为我国东部黑线姬鼠分为东北亚种(A .agrarius. mantchuricus) , 华北亚种(A.a. pallidior) 和长江亚种(A.a.ningpoensis) 提供了证据。     

用mtDNA序列探讨羚牛亚种分类(偶蹄目:牛科)

羚牛4个亚种在我国均有分布,且四川亚种和秦岭亚种是我国的特产动物,但对于分布于甘肃南部羚牛种群的亚种归属则有着不同的观点。本文就羚牛3个亚种的分类地位从线粒体DNA细胞色素b和D－Loop序列水平上进行探讨,分析结果基本支持形态学上的亚种划分,但甘肃南部羚牛种群在两个DNA片段上均与羚牛四川亚种存在着一定的差异,而与秦岭亚种则完全一致。结合羚牛地理分布特点,认为将分布于甘肃南部的羚牛种群划分为羚牛秦岭亚种可能更合适。     

华东地区地方鸡品种mtDNA控制区遗传多样性

为了探明我国华东地区地方鸡品种的遗传多样性和群体遗传结构,追溯其母系起源和进化过程,利用PCR技术扩增了11个地方鸡品种的线粒体DNA(mtDNA)控制区(D-loop)序列,并结合NCBI数据库中已发表的红色原鸡(Gallus gallus)D-loop区全序列,分析了它们的遗传多样性与亲缘关系,构建了11个品种与红色原鸡系统发生邻接树。结果表明:11个地方品种mtDNA D-loop区全长为1,231或1,232 bp,其中1,231 bp的序列有196条,1,232 bp的序列有123条,经过比对发现,两者在859 bp处存在单碱基缺失。11个地方品种319个个体共计检测到变异位点37个,总体单倍型多样度核苷酸多样度和平均核苷酸差异分别为0.901±0.009、0.00573±0.000001和6.833。按照鸡mtDNA单倍型分类通用标准,共包含35种单倍型,可以分为A、B、C和E共4个分支(单倍型群),分别包括11、10、9和5个单倍型。中介网络图中11个鸡品种也很明显地分成了4个支系,分别含有100、118、47和54条序列。系统发育树分为4个大枝,海南亚种(G.gallus jabouillei)自成一枝;C单倍型群与4个亚种的红色原鸡聚为一枝;E单倍型群与2个亚种红色原鸡聚为一枝;A和B单倍型群只与滇南亚种(G.gallus spadiceus)聚为一枝。11个品种中,除了狼山和丝羽乌骨鸡2个标准化品种外,都有很高的遗传多样性,可开发选择潜力很大。没有发现线粒体品种特异性DNA序列。华东地区地方品种至少有4个母系起源,部分品种可能受到了欧美高产品系的渗入。     

青岛近海路氏双髻鲨群体遗传学分析

研究采集了青岛近海23尾路氏双髻鲨(Sphyrna lewini), 通过线粒体DNA控制区片段对其遗传多样性进行分析。研究结果显示: 在23个个体的控制区序列上存在13个变异位点, 未检测到插入/缺失位点; 检测到7个单倍型, 其中3个为个体共享单倍型(Hap1、Hap3和Hap5), 4个为个体独有单倍型; 青岛近海路氏双髻鲨呈现中等水平的单倍型多样度和较低的核苷酸多样度; 与已报道的日照、霞浦群体间的遗传分化指数F_st值分别为–0.0571和–0.0328, 表明青岛群体与其他两个群体间不存在显著差异。以Sphyrna zygaena为外群构建NJ系统树显示本研究中7个单倍型共分成两支, 分别与来自太平洋、印度洋的单倍型类群聚类。中国近海的路氏双髻鲨作为一个具有较低遗传多样性的濒危物种, 其资源保护更应该引起足够的重视。     

Two Sympatric Phylogroups of the Chinese Water Deer (Hydropotes inermis) Identified by Mitochondrial DNA Control Region and Cytochrome b Gene Analyses

Mitochondrial DNA (mtDNA) control region (927 bp) and cytochrome b gene (1,140 bp) sequences of the Chinese water deer (Hydropotes inermis) from China and Korea were obtained to examine the taxonomic status of two subspecies, H. i. inermis from China and H. i. argyropus from Korea. Two sympatric mtDNA clades (a major clade from China and Korea and a minor clade from Korea) with an average genetic distance of 2.1% in the control region and 1.3% in the cytochrome b gene were detected. These findings are not consistent with the current classification by pelage color. We propose a reconsideration of the validity of the subspecies designation by the statistical comparison of morphological characters including body color. The major common mtDNA phylogroup in the two allopatric subspecies could be explained by the contiguous distribution of the Chinese water deer from east China to Korea until recent years. The restriction in the range and number of the Chinese subspecies after the last glacier might have caused the disappearance of the minor phylogroup in China. The taxonomic status of the two groups in Korea should be clarified using nuclear DNA marker analyses as well as morphological characters including pelage color.     

The complete mitochondrial genome of the striped field mouse, Apodemus agrarius (Rodentia, Murinae) from Korea

The striped field mouse, Apodemus agrarius, is the most common rodents in temperate forests of the Korean peninsula. We have determined the complete mitochondrial genome (JN629047) of A. agrarius from Korea. Apodemus agrarius mitogenome is 16,263 bp in length and has a base composition of 33.9% A, 29.9% T, 23.6% C, and 12.5% G. The total length of the 13 protein-coding genes is 11,398 bp long.     

Mitochondrial Cytochrome b Gene Sequence Diversity in the Korean Hare, Lepus coreanus Thomas (Mammalia, Lagomorpha)

Partial sequences of the mitochondrial cytochrome b gene of the Korean hare (Lepus coreanus) were analyzed to determine the degree of genetic diversity. Nine haplotypes were observed, and the maximum Tamura-Nei nucleotide distance among them was 2.8%, indicating that genetic diversity of L. coreanus is moderate. In order to clarify the Korean hare's taxonomic status and relationship with the Manchurian hare (L. mandshuricus) and the Chinese hare (L. sinensis), these nine haplotypes of the Korean hare were compared with 13 haplotypes from five other species of eastern Asian Lepus including L. mandshuricus and L. sinensis. The Korean hare was distinct in its cytochrome b gene, and it is confirmed that L. coreanus is a valid species, as noted by Jones and Johnson (1965, Univ. Kansas Publ. (Mus. Nat. Hist.) 16:357). Further analyses of mtDNA cytochrome b gene with additional specimens of L. coreanus from North Korea and other species of Lepus from eastern Asia are needed to clarify the taxonomic status of the divergent mtDNA clades of L. mandshuricus and L. sinensis.     

Phylogeography of the Alpine salamander, Salamandra atra (Salamandridae) and the influence of the Pleistocene climatic oscillations on population divergence

Fifty individuals of the endemic Alpine salamander, Salamandra atra, representing 13 populations throughout the range of the two currently recognized subspecies, atra and aurorae, were examined for sequence variation in a large portion (1050 bp) of the mitochondrial cytochrome b gene. We revealed a large number of mitochondrial DNA (mtDNA) haplotypes (10). Interpopulation sequence divergence was very low, ranging from 0 to 3.1%. The relationships among haplotypes were poorly resolved. The divergence time estimate between several mtDNA haplotypes suggested a pre-Pleistocene differentiation approximately 3 million years ago. Moreover, the impact of the Pleistocene glaciations on the phylogeographical patterns appears to have been secondary, although a somewhat reduced genetic variability was found in populations living in areas that were directly affected by the glaciation.     

Genetic diversity and phylogeography of Siberian roe deer,Caproulus pygargus,in central and peripheral populations

Current understanding of phylogeographical structure and genetic diversity of Siberian roe deer remains limited mainly due to small sample size and/or low geographical coverage in previous studies. Published data suggest at least two phylogroups: western (Ural Mountains and Western Siberia) and eastern (east from lake Baikal, including the Korean peninsula), but their phylogenetic relationship remains unclear. Combined sequences of cytochrome b (1140 bp) and the mtDNA control region (963 bp) were analyzed from 219 Siberian roe deer from 12 locations in Russia, Mongolia, and South Korea, which cover a large part of its range, to assess genetic diversity and phylogeographical status. Special emphasis was placed on the demographic history and genetic features of central, peripheral, and isolated populations. Results of median‐joining network and phylogenetic tree analyses indicate that Siberian roe deer from the Urals to the Pacific Ocean are genetically diverse and that geographical distribution and composition of haplogroups coincide with previously described ranges of the subspecies Capreolus pygargus pygargus and Capreolus pygargus tianschanicus. We found that peripheral populations in the northwestern parts of the species range (Urals), as well as the isolated population from Jeju Island, are genetically distinct from those in the core part of the range, both in terms of genetic diversity and quantitative composition of haplogroups. We also found that northwestern (Urals) and northern (Yakutia) peripheral populations share the same haplogroup and fall into the same phylogenetic clade with the isolated population from Jeju Island. This finding sheds light on the taxonomic status of the Jeju Island population and leads to hypotheses about the discordance of morphological and genetic evolution in isolated populations and specific genetic features of peripheral populations.     

Phylogeography of white-spotted Charr (Salvelinus leucomaenis) inferred from mitochondrial DNA sequences

The white-spotted charr (Salvelinus leucomaenis) is a coldwater-adapted fish distributed in far-eastern Asia. To assess phylogeographic patterns of this species over most of its range in the Japanese archipelago and Sakhalin Island, Russia, we examined nucleotide sequences of the mitochondrial DNA (mtDNA) cytochrome b region (557 bp) in 141 individuals from 50 populations. A total of 33 (5.5%) nucleotide positions were polymorphic and defined 29 haplotypes. Phylogenetic analysis assigned the observed haplotypes to four main clades, which were characterized by the idiosyncrasies and discontinuity of geographic distributions. The nested clade analyses revealed that the geographical distribution patterns of some haplotypes and clades were explained by historical event such as past fragmentation. Although substantial genetic differentiation was found among the four main clades, their geographic distributions overlapped extensively in several regions. Since white-spotted charr can potentially use both freshwater and marine environments, coexistence among different lineages can be attributed to secondary contact through range expansion by migratory individuals during multiple glacial periods after interglacial isolation. Finally, our data demonstrate that the current subspecies designation does not reflect the phylogeography of this species based on mtDNA analysis. Hierarchical analysis (AMOVA) also showed that genetic variation was far more pronounced within subspecies than among subspecies (i.e., among discrete regions). These results suggest that each population, rather than each subspecies, must be treated as an evolutionarily significant unit.     

Genetic differentiation of geographically separated populations of the southern green stink bug Nezara viridula (Hemiptera: Pentatomidae)

Genetic variation in the southern green stink bug Nezara viridula (Linnaeus) from 11 geographically separated sampling locations (Slovenia, France, Greece, Italy, Madeira, Japan, Guadeloupe, Galapagos, California, Brazil and Botswana) was studied by sequencing 16S and 28S rDNA, cytochrome b and cytochrome c oxidase subunit I gene fragments and random amplified polymorphic DNA (RAPD) analysis. Sequencing revealed 11 distinct haplotypes clustering into lineages A, B and C. Lineage C was characteristic for a single analysed specimen from Botswana. Lineage B was detected in Japan, and it probably arose in Asia. Haplotypes of European and American specimens belonged to lineage A; specimens from France, Slovenia, Madeira and Brazil shared highly similar haplotypes (>99%) from subgroup A1, while all the specimens from Greece, California, Galapagos and Guadeloupe shared a haplotype from subgroup A2. RAPD data were more variable but consistent with mtDNA sequences, revealing the same clustering. They separated the Botswanian specimen from Japanese specimens and from a group of more closely related specimens from Europe and America. Sequence and RAPD results both support the African origin of N. viridula, followed by dispersal to Asia (lineage B) and, more recently, by expansion to Europe and America (lineage A). RAPD analysis revealed two highly supported subgroups in Japan, congruent with mtDNA lineages A2 and B, suggesting multiple colonization of Japan. Invariant sequences at the 28S rDNA combined with other results do not support the hypothesis that cryptic (sibling) species exist within the populations investigated in this study.