Mitochondrial d-loop sequence variation and phylogeny of gobiobotine fishes

The mitochondrial DNA control region is amplified and sequenced from 8 genera and 10 species of gobiobotine fishes. The phylogenetic tree of Gobiobotinae and some representative species of other Cyprinid subfamilies obtained by the method of neighborhood joining, maximum likelihood and maximum parsimony with Danio rerio as an outgroup indicates that Gobiobotinae fishes are a monophyletic group which is close to Gobioninae subfamily. Gobiobotinae should be included into subfamily Gobioninae in terms of phylogenetic analysis. The research result supports that Gobiobotinae can be divided into genus Xenophysogobio and Gobiobotia. Xenophysogabio is the most primitive genera in the subfamily.

Mitochondrial cytochrome b sequence variations and phylogeny of the East Asian bagrid catfishes

The mitochondrial DNA cytochrome b gene was sequenced from 8 bagrid catfishes in China. Aligned with cytochrome b sequences from 9 bagrid catfishes in Japan, Korea and Russia retrieved from GenBank, and selected Silurus meridionalis, Liobagrus anguillicauda, Liobagrus reini and Phenacogrammus interruptus as outgroups, we constructed a matrix of 21 DNA sequences. The Kimura's two-parameter distances were calculated and molecule phylogenetic trees were constructed by using the maximum parsimony (MP) and neighbor-joining (NJ) methods. The results show that (i) there exist 3-bp deletions of mitochondrial cytochrome b gene compared with cypriniforms and characiforms; (ii) the molecular phylogenetic tree suggests that bagrid catfishes form a monophyletic group, and the genus Mystus is the earliest divergent in the East Asian bagrid catfishes, as well as the genus Pseudobagrus is a monophyletic group but the genus Pelteobagrus and Leiocassis are complicated; and 60 the evolution rate of the East Asian bagrids mitochondrial cytochrome b gene is about 0.18%-0.30% sequence divergence per million years.

线粒体 DNA 相关疾病的系统发育学研究

线粒体是细胞内提供能量的细胞器，并负责调节细胞的程序化死亡。因遗传 缺陷引起的线粒体功能障碍会导致ATP 合成障碍、能量产生不足而出现一系列 病症。线粒体DNA 相关疾病目前日益受到广泛的关注，然而在线粒体DNA 相 关疾病领域常用的病例对照法容易受到遗传背景、群体分层、数据质量等方面因 素的影响以致经常得到假阳性结果。系统发育分析方法有助于解决这些方面的问 题，因此我们以此分析方法开展了如下工作： 首先我们对线粒体DNA C1494T 突变及其所属单倍型类群和氨基糖苷类药 物性耳聋之间的关联进行了研究。之前有研究报道了两个中国氨基糖苷类药物性 耳聋的家系，经过对先征者的线粒体DNA 全序列测定在这两个家系中都发现了 C1494T 突变。我们采用系统发育方法分析这两个家系先征者的全序列后，发现 这两个个体都属于线粒体单倍型A。巧合的是，在我们之前的研究中，在一个来 自武汉的汉族样本WH6980 中也有C1494T 突变，而且该个体同样也属于单倍型 类群A。这不由得使人想到：C1494T 突变可能是单倍型类群A 中一个分支的界 定位点，或者，受母系遗传背景的影响，该突变偏好于在A 类群中发生。那么 很有可能单倍型类群A对能够引发氨基糖苷类药物性耳聋的C1494T 突变的发生 有促进作用。 为了验证这个假设，我们从三个省份随机选取了553 个正常个体来检测 C1494T 突变，以调查该突变在普通人群中的发生频率。另外，我们从1823 个中 国人样本中筛选出属于单倍型类群A 的111 个体，在这111 个个体中检测C1494T 突变，以调查该突变是否为单倍型类群A 特有的变异位点。我们的结果表明： 在553 个随机样本群中没有检测到C1494T 突变，这说明该突变是一个稀有的突 变，在正常人群中发生的频率极低。另外，在111 个属于单倍型类群A 的样本 群中，我们也没有检测到C1494T 突变的存在，这说明该突变并非单倍型类群A 特有的变异位点。经过对带有C1494T 突变的全序列进行综合的系统发育学分析 表明，这三条序列的C1494T 突变应是来源于同一次突变事件。同时，根据序列 之间共享变异位点的状况推断，两个耳聋家系具有很近的母系亲缘关系，我们推 测这两个家系的C1494T 突变是来源于一位共同的近期母系祖先。综合这些结果 表明，C1494T 突变的发生与单倍型类群即母系遗传背景没有相关性，是属于在人群中频率极低的散发性突变。 我们又将系统发育分析的方法应用于肿瘤相关的线粒体DNA 突变的研究 中。线粒体在细胞的自由基产生以及细胞凋亡中扮演重要角色；有研究报道线粒 体功能的缺陷能导致癌症的发生，同时又有很多报道指出癌变组织的线粒体基因 组存在异常。为探讨癌组织中线粒体基因组的变异情况及其在癌症的发生和发展 中所扮演的角色，我们选取乳腺癌早期患者为研究对象。 近年来，有大量的研究都报导了在癌组织中存在高频率高密度的线粒体 DNA 体细胞突变，并认为这些突变在肿瘤发生过程中可能具有功能相关性。但 这些研究存在着不可回避的问题，总结来讲归为三个方面：①数据质量差：很多 前面报道的突变数据，经系统发育分析，发现存在不少因样本交叉污染所造成的 假阳性突变。②所测片断太短。大多数研究只是检测了D-loop 区，只占了整个 基因组的约十六分之一，很难全面反映问题。③对照设置有问题，许多研究进行 简单的case-control 分析，忽略了不同个体间母系遗传背景的差异，导致大量假 阳性突变的产生。 针对以上问题的存在，我们基于系统发育分析的方法在中国的乳腺癌病人中 开展了一项研究。分别取得10 例乳腺癌早期患者的癌组织、癌旁组织、以及远 端正常组织；提取了总DNA，对每一份组织样品进行线粒体全基因组测序。这 样做有两个研究目的：第一是调查在严格的质量控制手段下，排除交叉污染所造 成的假阳性突变后，在癌症病人中是否仍然能观察到高频率高密度的线粒体 DNA 体细胞突变。第二个目的是调查中国乳腺癌患者线粒体DNA 体细胞突变情 况，为乳腺癌早期诊断提供有效的信息。 为了避免过去部分研究中出现的问题，我们采取了相应的措施。首先是在严 格的质量控制下，我们对同一个病人的三种不同组织，即癌组织，癌旁组织，正 常组织中的线粒体基因组进行全序列测定的方式来检测体细胞突变。同时将所测 得线粒体全基因组序列进行系统发育的分析；结果表明属于同一个病人的不同组 织都能忠实的聚到一支，而且每一条全序列都完整的带有所属单倍型特有的界定 位点，不存在任何样本交叉或者污染的情况；这样进一步确证了我们数据的可靠 性。同时经过这样的比较，可以非常清晰的筛选出在某一种组织类型中所发生的 体细胞突变。来自10 例病人的癌组织，癌旁组织和正常组织的29 条（7#患者远端正常组 织没有得到）线粒体基因组全序列中，我们只检测到了两个体细胞突变（T2275C 和A8601G）。这两个突变经PCR 克隆试验的验证，在远端正常组织中均不存在， 是真实的体细胞突变。突变A8601G 同时出现在3#患者的癌组织和癌旁组织中； 提示线粒体DNA 的体细胞突变可能先于细胞的癌变，这可能对乳腺癌的早期临 床诊断具有一定的意义。突变T2275C 只出现在6#患者的癌组织中，该突变位于 16S rRNA 基因非常保守的位点上，我们对该突变位点潜在的生物学作用作了进 一步的讨论。 在我们研究的病例中体细胞突变的比率要远远低于先前的报道；造成差异的 主要原因可能是因为我们采取了严格的质量控制手段。综合我们的研究结果表 明，先前所报道的乳腺癌组织中的线粒体基因体细胞突变的频率存在被高估的现 象。目前线粒体突变与肿瘤发生的关系还有待于更深入的研究，同时我们呼吁本 领域的研究要加强数据质量的可靠性。我们的研究表明，系统发育的分析方法在 线粒体DNA 相关疾病的研究中是有效的，因此我们推荐在线粒体DNA 相关疾 病的研究中进行推广。

印度人群线粒体 DNA 系统发育基因组学研究

人类线粒体DNA(mtDNA)是一个长度16,569bp 的环状分子，编码13 种蛋白 质、22 种tRNA 和2 种rRNA。由于mtDNA 全基因组信息具有缺乏重组、母系 遗传、高突变速率和相对较高的分辨率等特点，近年来已经成为重建人类历史的 重要工具。这些研究已经证实，mtDNA 最古老的六个单倍型类群，L0-L5，在非 洲特异的出现；而6-7 万年前从L3 衍生出的M 和N 两个超类群最终占领了世界 其他地区。然而，mtDNA 全序列研究在世界上某些特定地区尚是一片空白，其 中之一便是作为人类“走出非洲”的关键区域——印度。 为弥补这一空白，我们从 1200 个印度样品中选择了131 个可以代表所有主 要单倍型类群的个体，进行了全基因组扩增和测序，手工重建并软件验证了系统 发育关系树。我们的结果发现了12 个新的印度特有单倍型类群(N5, R7, R8, R30, R31, M34-M40)，修订了11 个已知特有单倍型类群(N1d, R5, R6, U2a, U2b, U2c, M2, M4, M5, M6, M30)的定义，详细描述了存在于印度的欧洲特有类群(HV, JT, U, N1, W)。 这一工作产生了多个推论。第一个是关于人类“走出非洲”假说长期以来 存在的争论。欧亚大陆和大洋洲mtDNA 在M 和N(包括R)超类群系统发育关系 上星状和不重叠的分布，表明了人类走出非洲是沿着亚洲海岸线(即所谓的“南 方路线”)的一个快速扩散的过程。第二个推论是关于存在于印度的欧洲特有世 系。与典型的欧洲世系相比，这些世系仅仅存在一到两个突变，从而证实了新石 器时代以来来自于近东新月地带或中亚高原的基因流。第三个推论涉及一个早期 的印度全序列研究。仔细分析其数据表明，他们的数据丢失了很多基部的特有突 变并产生了多个幻影突变，从而证实了系统发育思想对检测数据质量的作用。 随着印度人群 mtDNA 全序列研究的完成，人类mtDNA 系统发育的基本框 架得以建立。人类mtDNA 明显地呈现出大洲特异性分布。目前已经有两种假说用来解释这一现象。传统的观点把这一现象归于遗传漂变；而近期的选择假说认 为选择在人类mtDNA 的分化中扮演了极其重要的角色，而气候是主要的选择压 力。为解决这一争论，我们收集了来自南亚、大洋洲和东亚三个具有不同气候的 地区的mtDNA，使用直接计数的办法比较了各个大洲之间同义突变和异义突变 的差异。结果表明，几乎在所有的基因中，异义突变的数量低于同义突变的数量， 从而表明纯净化选择是人类mtDNA 进化中的主要力量。然而，在这三个大洲之 间没有发现显著的差异，表明mtDNA 在这三个区域上所承受的选择压力基本相 同。这一结果表明，气候不大可能是造成人类mtDNA 分化的主要原因。

中国都江堰汉族人群线粒体DNA 控制区遗传变异与长寿的相关性研究

随着社会的进步和医疗卫生水平的不断提高，人类获得了更高的平均寿 命，很多国家都步入了老龄化社会的行列。由于长寿具有遗传的倾向，所以 科学家们致力于人类长寿及衰老性疾病发生机理的研究，目的是为了使人类 在获得更长寿命的同时能够抵御或减缓老年性疾病的侵袭，远离衰老带来的 困扰，享受高质量的生活。 线粒体是真核生物的重要细胞器，具有长度为约为16569bp 的环状 DNA 分子。在人类群体，特别是欧洲群体的相关性研究中，线粒体DNA （mtDNA）编码区和控制区的一些多态性位点显示出与长寿及衰老性疾病的 相关。特别是mtDNA 控制区的C150T 变异除了在多个长寿人群中富集之外， 更是具有改变mtDNA 重链复制起始位点的功能。 为了探讨mtDNA 控制区多态性位点与中国汉族长寿人群是否存在相关 性，本研究在中国四川省都江堰地区采集了556 名年龄90 至108 岁的互无 关系的长寿老人血液样本，其中男性202 名，女性354 名。同时还采集了 214 名长寿老人的亲属和312 名无关对照的血液样本，年龄分别在10 至69 岁之间和22 到73 岁之间。我们对这些样本的mtDNA 通过测序和RFLP 等 手段进行了扫描，采集并记录了mtDNA 单倍型类群信息和控制区位点多态 信息。 在该人群中，本研究发现mtDNA 的主要单倍型类群与长寿没有显著的 相关性，总体单倍型类群频率分布在三个组别中基本一致（p=0.318）。对 mtDNA 控制区C150T 变异的频率在三个组别中做了包括总体频率差异，区 分mtDNA 单倍型类群的频率差异，区分样本性别的频率差异以及mtDNA 单倍型类群与性别信息联合的频率差异的分析。虽然在个别的比较中得到了 显著差异，但经过多重检验校正后，结果均变得不显著。此外，对146、152189 和195 等四个同样处于mtDNA 控制区复制起始区域的变异位点的初步 分析，同样没有获得显著的差异。不支持此前在欧洲长寿人群和日本长寿人 群得到的结论。 综上所述，本研究第一次在中国汉族人群中对mtDNA 控制区多态性与 长寿的相关性进行了研究。mtDNA 与人类长寿的关系还有待于更深层次的 机理性研究和功能性研究来揭示。

麂属动物的DNA进化

本文分析了麂属动物及其近缘种的线粒体DNA（Mitochondrial DNA, mtDNA)和核塘体DNA（Ribosomal DNA, rDNA）限制性片段长度多态性（RFLP），建立了麂属动物mtDNA和rDNA的限制性内切酶间谱。据此计算出各个物种的种内及种间的遗传距离，构建了麂属动物的种内及种间的分子聚类图。结果表明，在现生麂类中，黑麂和贡山麂之间的关缘关系最近，其次是费氏麂；印度麂是一个特化的特种，它和黑麂支系（包括费氏麂、贡山麂和黑麂）可能是从小麂的祖先类群中独立分化出来的。在近缘物种中，毛冠鹿与麂属动物的亲缘关系较近。结合前人有关的工作，计算19种鹿科动物之间的遗传距离并绘制它们的分子聚类图。

重离子束辐照诱变啤酒酵母线粒体DNA研究

摘要 II 5. 在不同注量离子束辐照后筛选出来的呼吸缺陷型酵母菌株中扩增获得位 于第 12 染色体上的 SOF1 基因，而在同样的扩增体系中没有得到野生型 菌株的该基因。 6. 选取离子束辐照后筛选出来的呼吸缺陷型酵母菌株再次进行辐照，发现 其在低剂量范围（＜0.93Gy）辐照下非常敏感，而在高剂量范围（＞ 0.93Gy）又表现出一定程度的辐射抗性。 结论： 1. 离子束辐照酵母细胞，直接或间接作用于酵母线粒体DNA，导致线粒体 DNA损伤，形成呼吸缺陷的酵母菌株。 2. I 类内含子和 II 类内含子对于离子束辐照的敏感性不同： II 类内含子比较 稳定，II 类内含子可能利用自身编码的反转录酶通过目的DNA引导的反 转录机制对受到辐照损伤的II 类内含子进行修复。 3. 离子束辐照后 SOF1 基因可能发生了突变，影响酵母细胞的生长。 4. 呼吸缺陷型酵母菌株因其线粒体 DNA发生变化及线粒体功能的改变， 使 呼吸缺陷型酵母菌株在不同剂量区的离子束辐照下表现不同辐射敏感 性。目的： 研究啤酒酵母的线粒体 DNA 在重离子辐照作用下的突变效应及其突变机 理。 材料与方法： 利用兰州重离子研究装置(HIRFL)加速的氖、碳离子辐照酵母细胞，用 TTC 显色培养基筛选呼吸缺陷型酵母菌株，并用 mtDNA 限制性酶切手段分析其突变 规律。采用 PCR扩增并对目的产物测序的方法对辐照后线粒体DNA上的 I 类内 含子和 II类内含子进行研究。 结果： 1. TTC 显色实验表明：离子束辐照导致酵母线粒体上的电子传递链发生改 变，产生的还原氢减少，造成呼吸缺陷。 2. 利用限制性酶切实验对线粒体 DNA进行研究，结果表明：离子束辐照诱 变筛选出来的呼吸缺陷型酵母菌株其线粒体DNA变化明显： 主要表现为 酶切条带缺失严重。即使在同一注量下筛选出来的呼吸缺陷型酵母菌株， 其酶切图谱也不相同。 3. 通过 PCR 手段对辐照后酵母线粒体 DNA 碱基序列进一步进行分析，发 现经不同注量离子束辐照后筛选出来的呼吸缺陷型酵母菌株，其I 类内含 子（ai4 and ai5）经设计不同引物进行扩增，没有获得目的条带，说明此 序列发生了突变，可能对离子束辐照比较敏感。 4. 经不同注量离子束辐照后筛选出来的呼吸缺陷型酵母菌株，其 II 类内含 子（ai2）的碱基序列与野生型相比没有变化，表现出在离子束辐照作用 下比较稳定的特性。

Differences in the rDNA-bearing chromosome divide the Asian-Pacific and Atlantic species of Crassostrea (Bivalvia, Mollusca)

Karyotype and chromosomal location of the major ribosomal RNA genes (rDNA) were studied using fluorescence in situ hybridization (FISH) in five species of Crassostrea: three Asian-Pacific species (C. gigas, C. plicatula, and C. ariakensis) and two Atlantic species (C. virginica and C. rhizophorae). FISH probes were made by PCR amplification of the intergenic transcribed spacer between the 18S and 5.8S rRNA genes, and labeled with digoxigenin-11-dUTP. All five species had a haploid number of 10 chromosomes. The Atlantic species had 1-2 submetacentric chromosomes, while the three Pacific species had none. FISH with metaphase chromosomes detected a single telomeric locus for rDNA in all five species without any variation. In all three Pacific species, rDNA was located on the long arm of Chromosome 10 (10q)-the smallest chromosome. In the two Atlantic species, rDNA was located on the short arm of Chromosome 2 (2p)-the second longest chromosome. A review of other studies reveals the same distribution of NOR sites (putative rDNA loci) in three other species: on 10q in C. sikamea and C. angulata from the Pacific Ocean and on 2p in C. gasar from the western Atlantic. All data support the conclusion that differences in size and shape of the rDNA-bearing chromosome represent a major divide between Asian-Pacific and Atlantic species of Crassostrea. This finding suggests that chromosomal divergence can occur under seemingly conserved karyotypes and may play a role in reproductive isolation and speciation.

The complete mitochondrial genome of the large yellow croaker, Larimichthys crocea (Perciformes, Sciaenidae): Unusual features of its control region and the phylogenetic position of the Sciaenidae

To understand the systematic status of Larimichthys crocea in the Percoidei, we determined the complete mitochondrial (mt) genome sequence using 454 sequencing-by-synthesis technology. The complete mt genome is 16,466 bp in length including the typical structure of 22 tRNAs, 2 rRNAs, 13 protein-coding genes and the noncoding control region (CR). Further sequencing for the complete CR was performed using the primers Cyt b-F and 12S-R on six L crocea individuals and two L polyactis individuals. Interestingly, all seven CR sequences from L crocea were identical while the three sequences from L polyactis were distinct (including one from GenBank). Although the conserved blocks such as TAS and CSB-1, -2, and -3 are readily identifiable in the control regions of the two species, the typical central conserved blocks CSB-D, -E, and -F could not be detected, while they are found in Cynoscion acoupa of Sciaenidae and other Percoidei species. Phylogenetic analysis shows that L crocea is a relatively recently emerged species in Sciaenidae and this family is closely related to family Pomacanthidae within the Percoidei. L crocea, as the first species of Sciaenidae with complete mitochondrial genome available, will provide important information on the molecular evolution of the group. Moreover, the genus-specific pair of primers designed in this study for amplifying the complete mt control region will be very useful in studies on the population genetics and conservation biology of Larimichthys. (c) 2008 Elsevier B.V. All rights reserved.

Characterization and mapping of expressed sequence tags isolated from a subtracted cDNA library of Litopenaeus vannamei injected with white spot syndrome virus

A large number of polymorphic simple sequence repeats (SSRs) or microsatellites are needed to develop a genetic map for shrimp. However, developing an SSR map is very time-consuming, expensive, and most SSRs are not specifically linked to gene loci of immediate interest. We report here on our strategy to develop polymorphic markers using expressed sequence tags (ESTs) by designing primers flanking single or multiple SSRs with three or more repeats. A subtracted cDNA library was prepared using RNA from specific pathogen-free (SPF) Litopenaeus vannamei juveniles (similar to 1 g) collected before (0) and after (48 h) inoculation with the China isolate of white spot syndrome virus (WSSV). A total of 224 clones were sequenced, 194 of which were useful for homology comparisons against annotated genes in NCBI nonredundant (nr) and protein databases, providing 179 sequences encoded by nuclear DNA, 4 mitochondrial DNA, and 11 were similar to portions of WSSV genome. The nuclear sequences clustered in 43 groups, 11 of which were homologous to various ESTs of unknown function, 4 had no homology to any sequence, and 28 showed similarities to known genes of invertebrates and vertebrates, representatives of cellular metabolic processes such as calcium ion balance, cytoskeleton mRNAs, and protein synthesis. A few sequences were homologous to immune system-related (allergens) genes and two were similar to motifs of the sex-lethal gene of Drosophila. A large number of EST sequences were similar to domains of the EF-hand superfamily (Ca2+ binding motif and FRQ protein domain of myosin light chains). Single or multiple SSRs with three or more repeats were found in approximately 61 % of the 179 nuclear sequences. Primer sets were designed from 28 sequences representing 19 known or putative genes and tested for polymorphism (EST-SSR marker) in a small test panel containing 16 individuals. Ten (53%) of the 19 putative or unknown function genes were polymorphic, 4 monomorphic, and 3 either failed to satisfactorily amplify genomic DNA or the allele amplification conditions need to be further optimized. Five polymorphic ESTs were genotyped with the entire reference mapping family, two of them (actin, accession #CX535973 and shrimp allergen arginine kinase, accession #CX535999) did not amplify with all offspring of the IRMF panel suggesting presence of null alleles, and three of them amplified in most of the IRM F offspring and were used for linkage analysis. EF-hand motif of myosin light chain (accession #CX535935) was placed in ShrimpMap's linkage group 7, whereas ribosomal protein S5 (accession #CX535957) and troponin I (accession #CX535976) remained unassigned. Results indicate that (a) a large number of ESTs isolated from this cDNA library are similar to cytoskeleton mRNAs and may reflect a normal pathway of the cellular response after im infection with WSSV, and (b) primers flanking single or multiple SSRs with three or more repeats from shrimp ESTs could be an efficient approach to develop polymorphic markers useful for linkage mapping. Work is underway to map additional SSR-containing ESTs from this and other cDNA libraries as a plausible strategy to increase marker density in ShrimpMap.

Mitochondrial and chloroplast phylogeography of Picea crassifolia Kom. (Pinaceae) in the Qinghai-Tibetan Plateau and adjacent highlands

The disjunct distribution of forests in the Qinghai-Tibetan Plateau (QTP) and adjacent Helan Shan and Daqing Shan highlands provides an excellent model to examine vegetation shifts, glacial refugia and gene flow of key species in this complex landscape region in response to past climatic oscillations and human disturbance. In this study, we examined maternally inherited mitochondrial DNA (nad1 intron b/c and nad5 intron 1) and paternally inherited chloroplast DNA (trnC-trnD) sequence variation within a dominant forest species, Picea crassifolia Kom. We recovered nine mitotypes and two chlorotypes in a survey of 442 individuals from 32 populations sampled throughout the species' range. Significant mitochondrial DNA population subdivision was detected (G(ST) = 0.512; N-ST = 0.679), suggesting low levels of recurrent gene flow through seeds among populations and significant phylogeographical structure (N-ST > GST, P < 0.05). Plateau haplotypes differed in sequence from those in the adjacent highlands, suggesting a long period of allopatric fragmentation between the species in the two regions and the presence of independent refugia in each region during Quaternary glaciations. On the QTP platform, all but one of the disjunct populations surveyed were fixed for the same mitotype, while most populations at the plateau edge contained more than one haplotype with the mitotype that was fixed in plateau platform populations always present at high frequency. This distribution pattern suggests that present-day disjunct populations on the QTP platform experienced a common recolonization history. The same phylogeographical pattern, however, was not detected for paternally inherited chloroplast DNA haplotypes. Two chlorotypes were distributed throughout the range of the species with little geographical population differentiation (G(ST) = N-ST = 0.093). This provides evidence for highly efficient pollen-mediated gene flow among isolated forest patches, both within and between the QTP and adjacent highland populations. A lack of isolation to pollen-mediated gene flow between forests on the QTP and adjacent highlands is surprising given that the Tengger Desert has been a geographical barrier between these two regions for approximately the last 1.8 million years.

Mitochondrial cytochrome b sequence variation and phylogenetics of the highly specialized schizothoracine fishes (Teleostei : Cyprinidae) in the Qinghai-Tibet Plateau

The complete 1140 bp mitochondial cytochrome b sequences were obtained from 39 individuals representing five species of all four genera of highly specialized schizothoracine fishes distributed in the Qinghai-Tibet plateau. Sequence variation of the cytochrome b gene was surveyed among the 39 individuals as well as three primitive schizothoracines and one outgroup. Phylogenetic analysis suggested that the group assignment based on 1140 bp of the cytochrome b sequence is obviously; different from previous assignments, and the highly specialized schizothoracine fishes (Schizopygopsis pylzovi, Gymnocypris przewalskii, G. eckloni, Chuanchia lablosa, and Platypharodon extremus) form a monophyletic group that is sister to the clade formed by the primitive schizothoracine fishes (Schizothorax prenanti, S. pseudaksaiensis, and S. argentatus). The haplotypes of Schizopygopsis pylzovi and G. przewalskii were paraphyletic based on cytochrome b data, which most likely reflected incomplete sorting of mitochondrial DNA lineages. The diploid chromosome numbers of Schizofhoracinae were considered in phylogenetic analysis and provided a clear pattern of relationships. Molecular dating estimated for highly specialized schizothoracine fishes suggested that the highly specialized schizothoracine fishes diverged in the late Miocene Pliocene to Pleistocene (4.5x10(4)-4.05x10(6) Years BP). The relationship between the cladogenesis of highly specialized schizothoracine fishes and geographical events of the Qinghai-Tibet plateau is discussed.

云南僰人源流的父系和母系遗传研究

对云南僰人32份男性DNA样本进行Y染色体单倍型以及mitochondrial DNA (mtDNA)单倍型分析,结果发现云南僰人的父系和母系遗传组分都表现出典型的南方人群的遗传特征.由僰人的数据结合已经发表的东亚人群的Y染色体和mtDNA单倍型(haplotype)数据进行Multidimensional Scaling(MDS)分析,结果表明,在MDS分布图中僰人群体的Y染色体单倍型和mtDNA单倍型都与南方人群聚在一起.这一结果支持僰人的遗传族源为东亚南方人群后裔,与考古学的推论相一致.结合历史和考古学证据来探讨僰人的起源和史前迁移,为揭开"僰人悬棺"这种独特的考古文化的起源和史前传播提供遗传学的研究证据.

Gene admixture in the Silk Road region of China: Evidence from mtDNA and melanocortin 1 receptor polymorphism

Mitochondrial DNA control region segment I sequences and melanocortin 1 receptor (MC1R) gene polymorphism were examined in ethnic populations in the silk road region of China. Both the frequencies of the MC1R variants and the results of mtDNA data in this region presented intermediate values between those of Europe and East and Southeast Asia, which suggested extensive gene admixture in this area and was in general agreement with previous studies. Phylogenetic analysis of the ethnic populations in the Silk Road region that based on mtDNA data didn't show expected cluster pattern according to their ethnogenesis. We suspect that a high migration rate in female among these closely related populations and other three demographic events might account for it.

食肉目哺乳动物的系统发育学研究概述

Carnivora; Caniformia; Feliformia; Phylogenetic tree; Mitochondrial DNA; Nuclear genes; 【摘要】 追溯生物界不同生物类型的起源及进化关系,即重建生物类群的系统发育树是进化生物学领域中一个十分重要的内容。食肉目哺乳动物位于食物链顶端,很多成员不仅在我国野生动物保护工作中占有重要地位,而且还是研究动物适应性进化遗传机制的重要模式生物。因而,食肉目物种作为物种资源中的一个重要类群,其系统发育学一直是国内外研究的热门课题。构建可靠的食肉目分子系统树,无疑将具有重要的进化理论意义和保护生物学价值。鉴于目前食肉目各科间系统发育关系仍然处于“广泛争论”的状态,本文将针对食肉目科水平上的系统发育学研究进展,包括来自于形态学特征、细胞学及分子生物学方面的证据,做简要概述,并提出目前研究中存在的问题。这对今后食肉目系统发育方面的进一步研究工作具有指导意义,并为以该类群作为模式生物开展适应性进化研究奠定基础