Low mtDNA Cytb diversity and shallow population structure of Eleutheronema tetradactylum in the East China Sea and the South China Sea

Eleutheronema tetradactylum is an economically important fish species in China water. To investigate the genetic diversity and describe population structure of it, an 1151 base pair (bp) fragment of the mitochondrial DNA Cytb sequence was analyzed in 120 individuals from four populations in the East China Sea and the South China Sea. A total of 16 haplotypes were defined by 24 variable nucleotide sites. High level of haplotype diversity and low nucleotide diversity were observed in all populations. The results of AMOVA detected that 89.44% of the genetic variation occurred within populations. Significant genetic differentiations were detected among populations (0.05097, P < 0.05), but no large-scale regional differences were detected. Analysis of neutral evolution and mismatch distribution suggested no recent population expansion happened. The present results provided new information for genetic assessment, fishery management and conservation of this species.     

三种鳖线粒体DNA细胞色素b基因序列的比较分析

对中华鳖、砂鳖和山瑞鳖线粒体DNA细胞色素b基因进行了引物设计、PCR扩增、序列测定和PCR-PFLP(Polymerase chain reaction-restriction fragment length polymorphism)分析。研究结果表明中华鳖、砂鳖与山瑞鳖线粒体DNACytb基因的序列全长相同,均为1140bp,其A、C、G、T含量相似,分别为378个(33.2%)、322个(28.2%)1、22个(10.7%)、318个(27.9%)、373个(32.7%)、324个(28.4%)、124个(10.9%)、319个(28.0%)、380个(33.3%)3、30个(28.9%)、127个(11.1%)、303个(26.7%)。同源性及序列差异率分析表明:中华鳖与砂鳖b基因序列的同源性为92.3%,中华鳖、山瑞鳖的为85.0%,砂鳖、山瑞鳖的为84.1%;中华鳖与砂鳖b基因核苷酸序列间的差异率为7.7%,中华鳖、山瑞鳖间的序列差异率为15.0%,砂鳖、山瑞鳖间的序列差异率为15.9%,而中华鳖、砂鳖、山瑞鳖各自个体间的序列差异率分别为2.37%、0.88%和0.18%,种间差异显著。用内切酶酶切分析其扩增产物,结果表明:用内切酶NdeⅠ可准确鉴别砂鳖,而用内切酶BamHⅠ则可准确鉴别山瑞鳖。内切酶NdeⅠ和BamHⅠ的联用分析,可使这三种鳖在分子水平都得到明确的鉴定。从三种鳖线粒体DNACytb基因核苷酸序列的显著差异和酶切位点的变化,可以进一步证明砂鳖是不同于中华鳖的鳖属一新种。     

Taxonomic status of the loaches Paracobitis vignai and P. rhadinaea (Teleostei: Nemacheilidae) in Iran

The crested loaches of the genus Paracobitis from the Sistan basin (Iran) are reviewed based on morphological and mitochondrial cytb and COI sequences characters to check the status of taxa in phylogenetic trees. Paracobitis rhadinaea (Regan, 1906) and P. vignai Nalbant & Bianco, 1998 were known only based on minor morphological differences (presence or absence of scales, caudal fin shape, colour pattern and fish size). We failed to find any diagnostic molecular and morphological characters between them. Therefore, we regard P. vignai as a junior synonym of P. rhadinaea.     

基于mtDNA Cyt b序列分析齐口裂腹鱼群体遗传多样性

研究测定了长江上游4个齐口裂腹鱼(Schizothorax prenanti)野生群体(重庆巫溪、重庆城口、四川雅安、四川阿坝)共104个个体的线粒体Cyt b基因部分序列, 以探讨齐口裂腹鱼野生群体的遗传多样性和遗传结构。结果表明: 在104个个体Cyt b序列中共检测到43个多态性位点, 25个单倍型。4个齐口裂腹鱼群体的单倍型多样性介于0.704—0.884, 核苷酸多样性介于0.007—0.012。群体间Kimura双参数遗传距离介于0.008—0.017, 其中四川雅安群体与四川阿坝群体间遗传距离最近, 基因交流频繁。重庆城口群体与四川雅安群体间遗传距离最远, 基因交流受阻。AMOVA分析表明, 齐口裂腹鱼的遗传分化主要来自群体内部, 且组群间、组群内群体间和群体内存在显著的遗传分化。中性检验得到Tajima’s D和 Fu’s F_s的值不显著, 且歧点分布图呈多峰, 表明长江上游4个齐口裂腹鱼野生群体未经历过种群扩张。研究旨为齐口裂腹鱼野生资源保护提供必要参考意见, 同时为齐口裂腹鱼种质资源合理开发和利用提供理论依据。     

以线粒体DNA Cytb确立红喉雉鹑和黄喉雉鹑的分类地位

采用PCR技术获得红喉雉鹑Tetraophasis obscurus和黄喉雉鹑Tetraophasis szechenyii等物种的828 bp Cytb基因片段,雉鹑属Tetraophasis可变位点为26个,并用MEGA3和K-Estimator 6.1软件进行了序列分析.研究发现红喉雉鹑和黄喉雉鹑之间序列差异为3.0%～3.1%,与其它属种间序列差异相比较,两种雉鹑之间的序列差异达到了种的水平;它们地理分布互相替代;形态差异稳定.地理分布、形态特征和遗传特征均验证了它们应为两个独立的种.依据分子进化速率推测,两种雉鹑分歧时间距今2.0Myr左右,与早更新世冰期发生的时间一致.雉鹑起源于青藏高原,随着青藏高原的抬升,早更新世冰期气候变冷,寒温性针叶林的林线下移,大面积高原草甸出现,雉鹑的祖先种群被隔离进化,而形成了两种雉鹑.     

The complete mitochondrial genome of the Sichuan taimen (Hucho bleekeri): repetitive sequences in the control region and phylogenetic implications for Salmonidae

The complete mitochondrial DNA genome of the Sichuan taimen (Hucho bleekeri) was determined by the long and accurate polymerase chain reaction (LA-PCR) and primer walking sequence method. The entire mitochondrial genome of this species is 16,997 bp in length, making it the longest among the completely sequenced Salmonidae mitochondrial genomes. It consists of two ribosomal RNA (rRNA) genes, 13 protein-coding genes, 22 transfer RNA (tRNA) genes, and one control region (CR). The gene arrangement, nucleotide composition, and codon usage pattern of the mitochondrial genome are similar to those of other teleosts. A T-type mononucleotide microsatellite and an 82 bp tandem repeat were identified in the control region, which were almost identical among the three H. bleekeri individuals examined. Both phylogenetic analyses based on 12 concatenated protein-coding genes of the heavy strand and on just the control region show that H. bleekeri is a basal species in Salmoninae. In addition, Salmo, Salvelinus and Oncorhynchus all represent monophyletic groups, respectively. All freshwater species occupied basal phylogenetic positions, and also possessed various tandem repeats in their mitochondrial control regions. These results support established phylogenetic relationships among genera in Salmonidae based on morphological and molecular analyses, and are consistent with the hypothesis that Salmonidae evolved from freshwater species.     

Molecular phylogeny of New World sheath-tailed bats (Emballonuridae: Diclidurini) based on loci from the four genetic transmission systems in mammals

Multiple unlinked loci are surveyed in a methodological approach for mammalian systematics that uses genes from the four pathways of genetic transmission: mitochondrial, autosomal, and X and Y sex chromosomes. Each of these components has different properties, such as effective population size, mutation rate, and recombination, that result in a robust hypothesis of evolutionary history. The utility of this experimental design is tested with bats in the family Emballonuridae and the hypothesis that the New World taxa are monophyletic. Parsimony and Bayesian analyses of the individual data sets give generally congruent topologies with high bootstrap proportions and posterior probabilities for monophyletic clades representing species and genera. The mitochondrial gene has significantly faster rates of substitution, higher levels of homoplasy, and a greater degree of saturation than the nuclear genes that contributed to the loss of phylogenetic signal at deeper branches of the tree. However, there is better resolution and support for the more slowly evolving nuclear introns including a New World clade, indicating a single origin of emballonurid bats in the Neotropics (tribe Diclidurini). One novel subtribe has a hard basal polytomy that is unresolved for all of the nuclear partitions, suggesting a rapid burst of evolution during the diversification of genera.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 93 , 189–209.     

基于线粒体COⅡ和Cytb基因序列的8种漠甲系统发育关系

对8种漠甲昆虫线粒体Cytb基因579bp和COⅡ基因585bp的序列片段进行联合分析。结果表明,8种甲虫在长度为1164bp的序列中碱基T,C,A,G的平均含量分别为33.7%,21.2%,33.5和11.6%,A+T平均含量明显高于G+C含量。密码子第三位点A+T含量高达77.2%,而该位点G的平均含量仅为4.0%。序列中碱基替换多发生于第三位点,转换略多于颠换,转换主要以C←T为主,颠换以A←T为主。以土甲族的Eumylada potanini为外群构建的系统发育树表明:漠王和漠甲以很高的置信值聚为一支,二者的关系与Bouchard最新的分类观点相吻合,即漠王族并入漠甲族成为一族;鳖甲族在支序图上另成一支,可视为一个单系群。     

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。     

基于Cytb基因的动物检材种属鉴定在非传统领域的部分运用

传统意义上的种属鉴定主要用于法医学研究方面,然而随着科技的进步及社会发展的需要,种属鉴定在其他领域也有广泛的应用。主要以线粒体Cytb基因为切入点,简要介绍了线粒体DNA及Cytb基因的结构和特点,总结和归纳了基于Cytb基因进行种属鉴定的相关技术方法在打击动物走私、濒危物种保护、食品及动物产品或制品等的检验检疫方面的运用情况,并对存在的问题及发展趋势做一阐述。