尖塘鳢属鱼类线粒体12SrRNA基因序列分析

利用PCR技术扩增和测序了线纹尖塘鳢、云斑尖塘鳢和海丰沙塘鳢线粒体12SrRNA基因,结合从GenBank中下载的部分同源序列,共分析了5种鱼类的系统发育关系。在Kimura2-parameter模型构建的邻接树中,原产泰国的云斑尖塘鳢与原产澳州线纹尖塘鳢均为单系类群,二者为亲缘关系最为密切的姐妹群,海丰沙塘鳢与其它群体的亲缘关系较远,支持将尖塘鳢属从塘鳢属中分出的传统分类处理。尖塘鳢属内云斑尖塘鳢和线纹尖塘鳢鱼类种内DNA序列无差异,而种间差异明显,表明线粒体12SrRNA基因可作为塘鳢科鱼类种类鉴定的良好分子标记。     

洞庭湖水系中华沙塘鳢的形态和核型研究

对取材于洞庭湖水系沅水和澧水的中华沙塘鳢Odontobutis sinensis进行了形态特征及染色体核型分析,并对其分类地位进行了探讨。染色体标本制作采用PHA和秋水仙素腹腔注射、肾细胞直接制片法。在形态上,洞庭湖水系中华沙塘鳢与其他水域沙塘鳢属鱼类既具相似性,又有各自特征;核型分析显示其二倍体染色体众数为2N=44,核型公式为8ST+36T,染色体臂数(NF)为44,与其他水域沙塘鳢属鱼类核型组成存在差异。     

浙江3个河川沙塘鳢群体线粒体16S rRNA基因序列的研究

为进一步了解浙江省河川沙塘鳢种质资源现状,通过对建德(JD)、钟管(ZG)的野生群体以及八里店养殖群体(BLD)的线粒体16S rRNA基因片段进行扩增和测定。获得了约590 bp的同源序列,其中变异位点72个,占分析位点的12.2%,含有66个简约信息位点,平均转颠换比值为1.83,4种碱基在所得序列中平均含量为A (30.8%)、G (21.1%)、T (23.6%)、C (24.5%),其中A+T含量为54.4%,略高于C+G含量。分子方差分析(AMOVA)显示,群体间的遗传分化系数(FST=0.393 47)在0.25以上,具有较高程度的遗传分化。基于邻接法构建的系统发育树和中间连接法构建的单倍型网络图均显示,建德群体单独聚为一支,且与另外两个群体相隔较远。群体中性检验、错配分析表明,钟管群体历史上较为稳定,未曾经历过显著种群扩张,建德群体近期经历过种群扩张。     

中华沙塘鳢在四川岷江汉阳水库发现

<正>2015年3月21日,在四川省岷江干流青神段进行水生生物多样性资源调查时,于汉阳电站坝上10 km瓮家乡(29°45.221'N,103°45.515'E,海拔450 m)采得16尾鱼类标本,经鉴定为中华沙塘鳢Odontobutis sinensis Wu,Chen et Chong,隶属于鲈形目Perciformes塘鳢科Eletridae沙塘鳢属。中华沙塘鳢主要见于珠江流域、海南岛及长江中上游的贵州、湖北、湖南及江西。标本现存于四川省农业科学院水产     

河川沙塘鳢对猎物种类和大小选择性研究

为了明确河川沙塘鳢(Odontobutis potamophila Günther)养殖宜投喂饵料生物的种类和规格, 通过室内外试验研究了沙塘鳢对生活习性不同8种饵料生物的种类选择, 对不同规格赤眼鳟(Squaliobarbus curriculus Richardson)的选择, 以及其不同生长阶段对饵料鱼大小的选择。研究结果表明, 沙塘鳢对麦鲮(Cirrhinus mrigala Hamilton)的选择指数显著大于其对剩余7种饵料生物的选择指数; 在无仿真水草条件下, 沙塘鳢对小规格饵料生物的选择指数显著大于对大、中规格饵料生物的选择指数; 在仿真水草条件下, 沙塘鳢对小规格饵料生物的选择指数显著大于其对大规格饵料生物的选择指数。在不同生长阶段中, 沙塘鳢全长与其捕食的饵料鱼全长呈现正相关关系; 沙塘鳢捕食的饵料鱼PPR值为0.23—0.73, 均值为0.49±0.1(均值±标准差); 随沙塘鳢规格的增加其捕食饵料鱼的PPR值降低, 捕食饵料鱼规格分布小于环境中饵料鱼规格分布。综上, 沙塘鳢养殖宜投喂饵料鱼麦鲮, 饵料鱼PPR值宜为0.38—0.6。     

基于线粒体COⅠ基因的沙鳅亚科鱼类DNA条形码及其分子系统发育研究

选择线粒体COⅠ基因作为分子标记,进行沙鳅亚科鱼类（Botiinae）DNA条形码及其分子系统发育研究。研究获得了沙鳅亚科7属19种共131个个体的COⅠ基因序列,利用MEGA5.0软件分析了沙鳅亚科鱼类COⅠ基因的序列特征,计算了种内及种间遗传距离。沙鳅亚科鱼类的分子系统发育关系的重建分别采用NJ法和Bayesian法。研究发现,沙鳅亚科COⅠ基因的碱基组成为: A 24.4%、T 29.5%、G 18.0%、C 28.1%。沙鳅亚科鱼类种内平均遗传距离为0.0020.000,种间平均遗传距离为0.1480.008。DNA条形码研究结果显示,所分析的19种沙鳅鱼类各自分别聚成单系分支,表明COⅠ基因在本研究中具有100%的物种鉴别率。同时,系统发育分析支持各属的单系性,并且结果显示沙鳅亚科鱼类聚为两个分支,其中一支由薄鳅属和副沙鳅属构成,另一分支则包括: （沙鳅属、色鳅属）和 中华沙鳅属、（缨须鳅属、安彦鳅属）。因此,COⅠ基因可以作为有效的分子标记对沙鳅亚科进行DNA条形码研究以及分子系统发育研究。       

基于16S rRNA和Cyt b基因序列的鰶亚科系统发育研究

对鰶亚科4属5种鱼类的线粒体基因组16S rRNA和Cyt b基因片段序列进行序列比较和系统发育关系分析。结果显示:5种鰶亚科鱼类的16S rRNA和Cy tb基因片段同源序列长度分别为525 bp和402 bp,序列联合后的序列总长度为927 bp,其中多态位点178个,简约信息位点123个。选取太平洋鲱Clupea pallasii和大西洋鲱C.harengus为外类群,采用邻接法(NJ)、最大简约法(MP)、最大似然法(ML)和贝叶斯法(BI)分别对2个基因片段序列进行了聚类分析,并联合2个基因片段利用邻接法、最大简约法和贝叶斯法进行分析。系统发育分析显示:斑鰶Konosirus punctatus与花鰶Clupanodon thriss亲缘关系最近,分布于美洲大陆的真鰶属Dorosoma鱼类与印度洋、太平洋分布的斑鰶属、花鰶属和海鰶属Nematalosa鱼类亲缘关系较远。     

河川沙塘鳢TLR2基因的克隆及表达分析

本研究以河川沙塘鳢(Odontobutis potamophila)为研究对象,采用RACE、实时荧光定量PCR等技术,首次对该鱼的TLR2基因进行了克隆和表达模式分析。主要研究结果如下:TLR2基因的c DNA全长序列为3 473 bp,包括2 631 bp开放阅读框(ORF)、75 bp的5'UTR区和767 bp的3'UTR区,并且具有脊椎动物典型的加尾信号AATAAA和26 bp的poly A尾巴,推测该序列编码876个氨基酸。蛋白结构预测表明TLR2蛋白的TIR结构域和LRR基序符合TLR家族的共同特征,其蛋白分子也存在多个功能位点。系统树中河川沙塘鳢与所有鱼类聚类为独立的分支,与鲈形目亲缘关系最近。应用RT-PCR法检测健康河川沙塘鳢鱼体中TLR2基因的m RNA组织表达差异情况。结果显示,TLR2基因在检测的12种组织中均有表达,在肝、肾、肠、血液和胃中都有较高的表达水平。人工感染嗜水气单胞菌后,对河川沙塘鳢3种主要免疫组织(脾、肾、肝)中进行不同时段表达量变化的检测,结果表明,TLR2基因在脾中感染后4 h表达水平发生下降,随后的24~48 h又上升,48 h时达到顶峰,72~96 h维持在略低于24 h水平上。在肾中,感染后表达水平持续上升,且24~96 h间保持相对稳定的表达水平。在肝组织中,感染后一直呈现上升趋势,72 h时达到顶峰,其他时段表达水平相差不大。可见TLR2在河川沙塘鳢抵御外源微生物侵染的先天免疫中发挥重要作用。     

基于线粒体Cyt b基因的中华沙塘鳢种群间遗传多样性分析

本研究以线粒体Cyt b基因为分子标记,对长江及珠江流域中华沙塘鳢(Odontobutis sinensis)的9个野生种群的遗传多样性进行分析。主要结论如下:(1)从108尾中华沙塘鳢样本中,共获得29个Cyt b基因单倍型(序列长度1 127 bp),总的单倍型多样性(Hd)、核苷酸多样性(π)值分别为0.929 0、0.009 41,呈现出较高的遗传多样性和较低的核苷酸多样性的特点;(2)中华沙塘鳢9种群间的FST值在0.110 6~0.998 8间(p0.01),K2-P遗传距离在0.002~0.022间,揭示各种群间存在显著的遗传分化;而多数种群间的基因交流值(N_m)小于1(p0.05),表明这些中华沙塘鳢种群间的基因交流有限。(3)中性进化检测和网络亲缘关系分析表明,中华沙塘鳢的桂林(GL)和洞庭湖(DTH)种群经历过种群扩张事件,推测中华沙塘鳢基因交流的方向为由长江水系种群流向珠江水系种群。通过以上研究为中华沙塘鳢的资源保护、开发与利用提供科学的理论依据。     

基于多变量形态度量学和线粒体Cytb序列的鲈属鱼类分类探讨

鲈属鱼类的分类在学术界尚存在很多争议。本文通过鲈属鱼类32个多变量形态学参数和1134bp的线粒体DNA细胞色素b序列的比较,对鲈属鱼类分类问题做了探讨。结果显示河鲈和伊犁鲈之间的形态距离为0.15,黄金鲈和伊犁鲈为0.14,河鲈和黄金鲈为0.09,在形态上黄金鲈和河鲈较接近,而伊犁鲈与前两者差异明显;主成分2(16.09%)对主成分1(21.71%)作图结果显示黄金鲈和河鲈有重叠区,而伊犁鲈与其它二种鲈有较大差距;细胞色素b同源序列差异百分比为河鲈与伊犁鲈13.08%、黄金鲈与伊犁鲈10.68%、黄金鲈与河鲈11.47%,鲈属鱼类间的碱基差异属于种间的遗传差异。MP、NJ和ML三种系统发育树在河鲈、黄金鲈和伊犁鲈三个种或亚种之间的拓扑结构一致,显示黄金鲈与伊犁鲈的演化关系较河鲈为近。根据20个样本的细胞色素b基因序列的遗传差异和系统发育树以及地理分布上的繁殖隔离,我们进一步认定黄金鲈和河鲈是不同的种,鲈属鱼类包括伊犁鲈、河鲈和黄金鲈三个种。     

鳜类系统发育的线粒体Cytb基因全序列分析

测定了鳜、大眼鳜、斑鳜、暗鳜、波纹鳜、长体鳜、中国少鳞鳜等7种鳜类12个个体的线粒体细胞色素b基因全序列。结合GenBank中的同源序列,共分析了9种鳜类的系统发育关系。序列分析表明,鳜属鱼类属内种间的遗传距离(0.015～0.093)明显小于少鳞鳜属鱼类属内种间的遗传距离(0.152～0.178)。在分子系统发育树上,长体鳜与鳜属的鳜、大眼鳜、斑鳜、波纹鳜、暗鳜聚合成一分支,少鳞鳜属的种类聚成另一分支;支持将长体鳜归入鳜属,鳜类分为鳜属和少鳞鳜属等二个属的分类处理。在鳜属鱼类中,鳜和大眼鳜亲缘关系十分密切;斑鳜与波纹鳜亲缘较近;长体鳜与鳜属其它5个种的亲缘关系较远。在少鳞鳜属鱼类中,中国少鳞鳜和日本少鳞鳜的亲缘关系较远,韩国少鳞鳜的系统位置较不明确。鳜类的单系性及其鳜类的系统位置仍有待进一步研究。     

A molecular phylogeny of the Percidae (Teleostei, Perciformes) based on mitochondrial DNA sequence

The family Percidae is among the most speciose families of northern hemisphere fishes with > 178 178 North American species and 14 Eurasian species. Previous phylogenetic studies have been hampered by a lack of informative characters, inadequate taxonomic sampling, and conflicting data. We estimated phylogenetic relationships among 54 percid species (9 of 10 genera and all but one subgenus of darters) and four outgroup taxa using mitochondrial DNA data from the 12S rRNA and cytochrome b genes. Four primary evolutionary lineages were consistently recovered: Etheostomatinae (Ammocrypta, Crystallaria, Etheostoma, and Percina), Perca, Luciopercinae (Romanichthys, Sander, and Zingel), and Gymnocephalus. Except Etheostoma and Zingel, all polytypic genera were monophyletic. The Etheostoma subgenus Nothonotus failed to resolve with other members of the genus resulting in a paraphyletic Etheostoma. The subfamily Percinae (Gymnocephalus and Perca) was not recovered in phylogenetic analyses with Gymnocephalus sister to Luciopercinae. Etheostomatinae and Romanichthyini were never resolved as sister groups supporting convergent evolution as the cause of small, benthic, stream-inhabiting percids in North American and Eurasian waters.     

Molecular phylogenetics and ecological diversification of the transisthmian fish genus Centropomus (Perciformes: Centropomidae).

Phylogenetic relationships among the 12 recognized fish species in the New World genus Centropomus (Pisces, Centropomidae) were analyzed using allozyme electrophoresis and 618 bp of the mitochondrial DNA 16S ribosomal RNA (rRNA) gene. Molecular phylogenetic trees were generally consistent with previously published partial hypotheses based on morphological evidence. However, previously undefined sister group relationships between major species groups were resolved using molecular data, and phylogenetic hypotheses for Centropomus based on 16S rRNA sequences were better supported than were allozyme-based hypotheses. The high level of congruence among the trees inferred from the nuclear and mitochondrial characters provided a firm phylogenetic basis for analysis of ecological diversification and molecular evolution in the genus. Compared to basal Centropomus species, members of the most nested species group were significantly larger in body size and occupied a marine niche only peripherally utilized by their congeners. We also observed substitution rate heterogeneity among 16S rRNA lineages; in contrast to expectations based on "metabolic rate" and "generation interval" models, relative substitution rates were faster than expected for the group of large-bodied snooks. Using the Pliocene rise of the Central American isthmian marine barrier to calibrate rates of 16S ribosomal gene evolution in Centropomus, we found that the rates for the genus were similar to those reported for higher vertebrates. Analysis of the three sets of transisthmian geminate taxa in Centropomus indicated that two of the pairs were probably formed during the Pliocene rise of the isthmus while the third pair diverged several million years earlier.     

Re-examining the phylogeny of clinically relevant Candida species and allied genera based on multigene analyses

Yeasts of the artificial genus Candida include plant endophytes, insect symbionts, and opportunistic human pathogens. Phylogenies based on rRNA gene and actin sequences confirmed that the genus is not monophyletic, and the relationships among Candida species and allied teleomorph genera are not clearly resolved. Protein-coding genes have been useful to resolve taxonomic positions among a broad range of fungi. Over 70 taxa of the genus Candida and its allied sexually reproducing genera were therefore selected, and their phylogenetic relationships were investigated using nuclear sequences of the largest subunit and second largest subunit of RNA polymerase II gene, actin, the second subunit of the mitochondrial cytochrome oxidase gene, and D1/D2 LSU rRNA gene. The DNA sequences were analysed by maximum parsimony and Bayesian inference, resulting in the recognition of six major phylogenetic groups (A-F). Group A contains six facultative pathogenic Candida species, which seem to have derived from nonpathogenic species, while Group B contains species of Clavispora, Metschnikowia, and Pichia guilliermondii. Species of Debaryomyces form an independent group C that is related to groups A and B. Pichia fermentans and other environmental species are concentrated in Group D. Group E, containing Pichia anomala, may be a sibling to group F, which is represented by the Saccharomyces species complex.     

Phylogenetic analysis of ticks (Acari: Ixodida) using mitochondrial genomes and nuclear rRNA genes indicates that the genus Amblyomma is polyphyletic

Our understanding of the phylogenetic relationships among tick lineages has been limited by the lack of resolution provided by the most commonly used phylogenetic markers. Mitochondrial genomes are increasingly used to address controversial phylogenetic relationships. To date, the complete mitochondrial genomes of eleven tick species have been sequenced; however, only three of these species are metastriate ticks, the most speciose lineage of ticks. In this study, we present the nucleotide sequences of the complete mitochondrial genomes of five more species of metastriate ticks: Amblyomma elaphense, Amblyomma fimbriatum, Amblyomma sphenodonti, Bothriocroton concolor and Bothriocroton undatum. We use complete mitochondrial genome sequences to address the phylogenetic placement of two morphologically 'primitive' species -Am. elaphense and Am. sphenodonti - with respect to the genus Amblyomma. Our analysis of these five mitochondrial genomes with the other eleven tick mitochondrial genomes, as well as analysis of nuclear rRNA genes, provides strong evidence that the genus Amblyomma is polyphyletic with the inclusion of Am. sphenodonti and Am. elaphense. A new genus or two new genera may be required to describe Am. sphenodonti and Am. elaphense. It is also possible that these two species are sisters to two established genera, Bothriocroton in the case of Am. sphenodonti, and Haemaphysalis in the case of Am. elaphense. However, other arrangements of these taxa cannot be excluded with the current data. Thus, while Am. sphenodonti and Am. elaphense do not belong in the genus Amblyomma, the phylogenetic placement of these two species cannot be resolved without more data from metastriate ticks, either greater sampling of mitochondrial genomes, or a large data set of nuclear genes.     

Complete nucleotide sequence of Japanese flounder (Paralichthys olivaceus) mitochondrial genome: structural properties and cue for resolving teleostean relationships

We cloned and sequenced the complete mitochondrial genome of Japanese flounder (Paralichthys olivaceus). A circular 17,090 bp mitochondrial genome from the flounder contains 37 structural genes as in other vertebrates so far reported. This is the first report of the complete mitochondrial sequence from a higher teleostean fish (Acanthopterygii). The organization including gene order is quite similar to that of other teleostean fishes as well as placental mammals. The putative control region of the Japanese flounder mitochondrial genome contains a length variable region of about a 74 bp tandem repeat cluster. As a preliminary study we adopted the maximum likelihood and neighbor-joining inference methods to examine phylogenetic relationships among teleostean and related fishes. Comparisons of amino acid sequences of protein-coding genes and nucleotide sequences of tRNA genes resolved some middle to deep branches among some teleostean fishes. The flounder mitochondrial genome does not show an indication of evolutionary rate difference among teleosts leading to difficulty in phylogenetic analyses, and our data is useful for future evolutionary studies dealing with higher teleostean fishes.     

First data on the molecular phylogeography of scincid lizards of the genus Mabuya

A 487-bp fragment of the mitochondrial 16S rRNA gene was sequenced in 26 species of the circumtropical lizard genus Mabuya and used to analyze phylogenetic relationships within the genus. The species from Africa and Madagascar formed a monophyletic group relative to the included Asian and South American taxa. The Malagasy species included (M. elegans, M. cf. dumasi, and M. comorensis) did not appear as a monophylum. Combined and separate analysis of the 16S data and additional sequences of the mitochondrial 12S rRNA, ND4, and cytochrome b genes (a total of 2255 bp) in one Asian, two Malagasy, and two African species also did not result consistently in a monophyletic grouping of the Malagasy taxa. However, a monophylum containing African and Malagasy taxa was strongly supported by the combined analysis. These preliminary results indicate that Mabuya probably colonized Madagascar from Africa through the Mozambique Channel.     

Reevaluation of phylogeny in the Drosophila obscura species group based on combined analysis of nucleotide sequences.

The Drosophila obscura species group has served as an important model system in many evolutionary and population genetic studies. Despite the amount of study this group has received, some phylogenetic relationships remain unclear. While individual analysis of different nuclear, mitochondrial, allozyme, restriction fragment, and morphological data partitions are able to discern relationships among closely related species, they are unable to resolve relationships among the five obscura species subgroups. A combined analysis of several nucleotide data sets is able to provide resolution and support for some nodes not seen or well supported in analyses of individual loci. A phylogeny of the obscura species group based on combined analysis of nucleotide sequences from six mitochondrial and five nuclear loci is presented here. The results of several different combined analyses indicate that the Old World obscura and subobscura subgroups form a monophyletic clade, although they are unable to resolve the relationships among the major lineages within the obscura species group.     

从线粒体16S rDNA序列探讨绒螯蟹类的系统发生关系

测定了绒螯蟹类各物种的线粒体16SrDNA部分片段的序列,构建了NJ树、ML树和MP树。序列歧异数据比较和各系统发生树都支持新绒螯蟹属(Neoeriocheir)为一个独立的属。在3种系统发生树中,直额绒螯蟹(Eriocheir recta)都是绒螯蟹属(Eriocheir)所有其它成员的姐妹群,并且广东珠江1只直额绒螯蟹标本的16SrDNA部分序列与台湾产台湾绒螯蟹(Eriocheir formasa)的相应序列相同。这些结果不支持平绒螯蟹属(Platyeriocheir)是一个有效的属,并表明E．formosa是E．recta的同物异名。绒螯蟹属(Eriocheir)所有其它成员聚为一个单系的分支,支持中华绒螯蟹、合浦绒螯蟹与日本绒螯蟹属于同一个物种Eriocheir japonica。16SrDNA部分序列的比对表明,产于台湾的日本绒螯蟹的此段序列与合浦绒螯蟹的相同,产于崇明岛的和产于美国旧金山海湾的中华绒螯蟹的此段序列与中华绒螯蟹单元型B的序列相同。