基于细胞色素b序列的鲹科分子系统发育

测定了中国鲹科8属9种鱼的细胞色素b基因的全序列(1141bp),结合来自GenBank中分布于美国、安哥拉、希腊以及巴拿马的鲹科4属14种鱼的相应同源序列生成供系统发育分析的序列矩阵,用最大简约法和邻接法构建分子系统树。结果显示:(1)支持科下设四个亚科(鲹亚科,亚科,鲳鲹亚科,鰆鲹科)阶元的分类系统;(2)亚科属下不宜设亚属分类阶元;(3)及达副叶鲹与丽叶鲹亲缘关系较近,应同属于副叶鲹属;(4)我国传统的鱼类检索系统将狮鼻鲳鲹误鉴定为卵形鲳鲹,建议予以修正。     

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。     

Molecular phylogenetic relationships of China Seas groupers based on cytochrome b gene fragment sequences

The classification and evolutionary relationships are important issues in the study of the groupers. Cytochrome b gene fragment of twenty-eight grouper species within six genera of subfamily Epinephelinae was amplified using PCR techniques and the sequences were analyzed to derive the phylogenetic relationships of the groupers from the China Seas. Genetic information indexes, including Kimura-2 parameter genetic distance and Ts/Tv ratios, were generated by using a variety of biology softwares. With Niphon spinosus, Pagrus major and Pagrus auriga as the designated outgroups, phylogenetic trees, which invoke additional homologous sequences of other Epinephelus fishes from GenBank, were constructed based on the neighbor-joining (NJ), maximum-parsimony (MP), maximum-likelihood (ML) and minimum-evolution (ME) methods. Several conclusions were drawn from the DNA sequences analysis: (1) genus Plectropomus, which was early diverged, is the most primitive group in the subfamily Epinephelinae; (2) genus Variola is more closely related to genus Cephalopolis than the other four genera; (3) genus Cephalopolis is a monophyletic group and more primitive than genus Epinephelus; (4) Promicrops lanceolatus and Cromileptes altivelis should be included in genus Epinephelus; (5) there exist two sister groups in genus Epinephelus.     

基于线粒体Cyt b基因部分序列的中国东方蜜蜂不同地理种群的系统发育

从线粒体DNA水平上对中国境内东方蜜蜂不同地理种群的系统发育进行研究,为保护和合理利用这一宝贵的蜂种资源提供理论基础。对我国10个省市的21群东方蜜蜂mtDNA Cyt b基因片段进行了扩增、测序,并以意大利蜂、卡尼鄂拉蜂、印尼蜂作为外群进行序列分析。采用Minimum-Spanning Network方法构建系统进化树。结果显示：扩增片段长度429bp;在得到的21条同源序列中,共检测出13个变异位点,其中转换数为9,颠换数为4,并且变异位点大都发生在密码子的第三位,无碱基插入或缺失;聚类分析结果显示西方蜜蜂、印尼蜂和东方蜜蜂各为独立的蜂种;在中国东方蜜蜂群体中,吉林、海南和云南的东方蜜蜂各为一个独立的类群,其它地区的东方蜜蜂为一个类群[动物学报54（6）：1005-1013,2008]。     

Identification of mosquito bloodmeals using mitochondrial cytochrome oxidase subunit I and cytochrome b gene sequences

Abstract.  Primer pairs were designed and protocols developed to selectively amplify segments of vertebrate mitochondrial cytochrome oxidase subunit 1 (COI) and cytochrome b (Cyt b ) mtDNA from the bloodmeals of mosquitoes (Diptera: Culicidae). The protocols use two pairs of nested COI primers and one pair of Cyt b primers to amplify short segments of DNA. Resultant sequences are then compared with sequences in GenBank, using the BLAST function, for putative host identification. Vertebrate DNA was amplified from 88% of our sample of 162 wild-caught, blood-fed mosquitoes from Oregon, U.S.A. and GenBank BLAST searches putatively identified 98% of the amplified sequences, including one amphibian, seven mammalian and 14 avian species. Criteria and caveats for putative identification of bloodmeals are discussed.     

乔氏新银鱼基于细胞色素b序列的种群遗传结构和种群历史

为促进乔氏新银鱼(Neosalanx jordani)种质资源的保护, 采集了长江流域和淮河流域5个乔氏新银鱼地理种群计129个样本, 利用线粒体细胞色素b基因(Cyt b)全序列作为分子标记, 初步分析了乔氏新银鱼种群的遗传多样性、遗传结构及种群历史动态。研究结果共检测到18个Cyt b单倍型, 发现和其他鱼类相比, 乔氏新银鱼具有较高的单倍型多样性(h, 0.590±0.047), 但核苷酸多样性较低(π, 0.00088±0.00011)。分子变异分析(AMOVA)表明, 乔氏新银鱼5个地理种群内个体间和流域内种群间均存在显著的遗传差异, 而长江流域与淮河流域之间遗传分化不明显, 显示乔氏新银鱼遗传分化与当前水系的分布格局不吻合。结果表明乔氏新银鱼目前的遗传格局主要是由于长距离独立的建群事件、基因流限制以及种群的持续扩张的共同作用而形成的。种群历史动态分析结果显示乔氏新银鱼种群为近期扩张种群, 其大部分变异发生在1.897万年之内, 与最后一次冰期全面消退、海平面上升、长江流域及淮河流域中下游大量湖泊(适宜生境)形成的时间相吻合。建议对现存的乔氏新银鱼种群, 特别是遗传多样性较高的那些种群(如鄱阳湖、太湖及洪泽湖种群)分别保护。     

Reproduction and morphology of Central American and Tres Marias spinner dolphins in the eastern tropical Pacific

Four geographic forms of spinner dolphin (Stenella longirostris) occur in the eastern tropical Pacific. Two of the forms are pelagic: the eastern and whitebelly, and two are coastal: the Central American and Tres Marias spinner dolphins. Of these, only the Tres Marias form lacks recognition appropriate for developing management and conservation plans. Vertical aerial photographs of 57 spinner dolphin schools taken between 1988 and 2006 were used to examine the range of coastal forms and to characterize their calving seasons. On the basis of adult body length, spinner dolphins photographed off Guerrero, Mexico, were identified as those of the Central American form, which is north of their known range, and those photographed farther north, approximately 112 km off Nayarit, Mexico, were identified as the Tres Marias form. Calving peaked from October through March, and there was evidence that calving seasons differed among geographic forms with coastal forms calving later than pelagic forms. We present the first life history parameter estimates for the coastal forms of spinner dolphins, provide additional support for recognition of the Tres Marias form, and the first evidence of the Central American form occurring north of the Gulf of Tehuantepec, Mexico.     

PCR-RFLP analysis of the cytochrome b gene in horse mitochondrial DNA

The mitochondrial DNA sequence of cytochrome b gene in a Thoroughbred horse was determined. By comparing DNA sequences between the Thoroughbred and published sequence data (two horses and one Grevyi zebra), polymerase chain reaction (PCR) primers were designed for amplification of a 590 bp DNA fragment in the cytochrome b gene, and PCR-restriction fragment length polymorphism (RFLP) analysis was studied in 140 horses of six breeds using three restriction enzymes ( AciI, BamHI, RsaI ). Two morphs were found using each of the three enzymes. By combining three enzymes morphs, the 140 horses examined were classified into four types. Type 2 was most frequent in all breeds.     

Phylogenetic relationships of Eurasian and American cyprinids using cytochrome b sequences

Neighbour-joining and parsimony analyses identified five lineages of cyprinids: (1) European leuciscins (including Notemigonus )+North American phoxinins (including Phoxinus phoxinus ); (2) European gobionins+Pseudorasbora ; (3) primarily Asian groups [cultrins+acheilognathins+ gobionins (excluding Abbotina )+xenocyprinins]; (4) Abbottina+Sinocyclocheilus+Acrossocheilus ; (5) cyprinins [excluding Sinocyclocheilus and Acrossocheilus ]+barbins+labeonins. Relationships among these lineages and the enigmatic taxa Rhodeus were not well-resolved. Tests of monophyly of subfamilies and previously proposed relationships were examined by constraining cytochrome b sequences data to fit previous hypotheses. The analysis of constrained trees indicated that sequence data were not consistent with most previously proposed relationships. Inconsistency was largely attributable to Asian taxa, such as Xenocypris and Xenocyprioides . Improved understanding of historical and taxonomic relationships in Cyprinidae will require further morphological and molecular studies on Asian cyprinids and taxa representative of the diversity found in Africa.     

A mtDNA phylogeny of sea eagles (genus Haliaeetus) based on nucleotide sequences of the cytochrome b-gene

The mitochondrial cytochrome b gene of seven species of sea eagles H. albicilla, H. leucocephalus, H. leucoryphus, H. leucogaster, H. sanfordi, H. pelagicus and H. vocifer was amplified by PCR and sequenced (1026 bp). Phylogeny reconstructions by the Maximum Parsimony and Neighbour-Joining methods produced similar trees in which sea eagles represent a monophyletic group. In addition, the clade H. albicilla/H. leucocephalus groups with the clade H. pelagicus/H. leucoryphus in a monophyletic boreal group while H. vocifer clusters with the H. leucogaster/H. sanfordi clade in a monophyletic tropical clade. The nearest relatives of sea eagles are the kites (genus Milvus) and buzzards (genus Buteo), whereas ‘booted’ eagles (genus Aquila) and vultures (genera Gyps and Aegypius) have diverged earlier from the accipitrid branch. Honey bussards (genus Pernis) and vultures of the genera Gypaetus and Neophron represent basal taxa of the accipitrid lineage. Falcons, New World vultures and the secretary bird (Sagittarius) appear in separate clades outside the Accipitridae.     

Comparative phylogeography of Zacco platypus and Opsariichthys bidens (Teleostei, Cyprinidae) in China based on cytochrome b sequences

We compared the phylogenetic relationships and the population structure of the sympatric cyprinids Opsariichthys bidens and Zacco platypus across the Yangzte, Pearl and Hai He drainages in Mainland China. We used the mitochondrial cytochrome b gene to evaluate their phylogeographical structure and to discuss further systematic implications. We sequenced 71 new individuals of Z. platypus to add to a previous study and to construct a geographically comparable data set with O. bidens (Mol. Phylogenet. Evol. 31, 2004, 192; Mol. Phylogenet. Evol. 37, 2005, 920). Phylogeographic analysis of Z. platypus identified two additional mtDNA lineages (Zacco E and F) in addition to the previously recovered lineages (Zacco A–D). The geographical range of the Zacco A was extended across the Pearl drainage, broadening considerably the previous distribution that was circumscribed to the Yangtze drainage. Our results show high molecular divergence between all Zacco A–F mtDNA lineages (mean TrN + I + G distance 17.7%, range 4.4–30.8%), and the population analyses showed that most of the genetic variation was found among major river tributaries (90.2%). Values of θ_ST higher than 0.97 corroborated this interruption of gene flow between Zacco A–F mtDNA lineages. Nested clade analysis results were also concordant with the phylogenetic relationships recovered. We found similar phylogeographic patterns in Z. platypus and O. bidens: they exhibited two widespread and common lineages (Zacco A and Opsariichthys 1) ranging across the Yangtze and Pearl drainages, and both taxa had Northern lineages (Zacco F and Opsariichthys 5) clearly differentiated. Similar phylogeographic patterns suggest that common historical factors have shaped current Zacco and Opsariichthys diversity. High genetic distances and θ_ST values suggested that differentiation of Zacco A–F mtDNA lineages and Opsariichthys 1–5 was not recent and was probably favoured by old geological events. However, population parameters and low genetic differentiation inside Zacco A and Opsariichthys 1 suggest a recent gene flow across river drainages. Mitochondrial‐based phylogenies for the combined data sets of Zacco A–F and Opsariichthys 1–5 mtDNA lineages always recovered both taxa as paraphyletic, highlighting the notion that their current systematics need a revision.     

基于线粒体Cyt b基因的全长序列探讨闭壳龟类的系统进化

采用PCR技术对淡水龟科具闭壳结构的黄缘盒龟、黄额盒龟、金头闭壳龟、潘氏闭壳龟、锯缘龟和白腹摄龟的线粒体Cytb基因的全长序列进行了PCR扩增和序列测定,并结合GenBank中16种淡水龟科物种的同源序列,进行了序列变异和系统发生分析。经C lustalX1.8软件对位排列后共有1154个位点,其中可变位点413个,简约信息位点301个;A＋T的平均含量（56.5%）高于G＋C（43.5%）。在氨基酸密码子中,第一位富含A,第二位富含T,第三位富含C;碱基转换/颠换率为5.97,碱基替换多发生在密码子第三位。以中华鳖和马来鳖为外群,通过最大简约法,最大似然法和贝叶斯法重建了分子系统树,均具有一致的拓扑结构,结果表明：金头闭壳龟和潘氏闭壳龟最先聚成一支,再和三线闭壳龟聚成一组,说明形态上相似的三种闭壳龟亲缘关系最近;闭壳龟属、盒龟属和单种锯缘龟属聚成一个单系的闭壳龟群,建议合并为闭壳龟属;齿缘龟属和果龟属聚为一支,它们与新的闭壳龟属关系较远,揭示闭壳结构的形成不是由一个共同祖先分化而来;乌龟属、花龟属和拟水龟属三属为并系起源,建议三属可以合并为一属。     

Primers and polymerase chain reaction conditions for DNA barcoding teleost fish based on the mitochondrial cytochrome b and nuclear rhodopsin genes

This report describes a set of 21 polymerase chain reaction primers and amplification conditions developed to barcode practically any teleost fish species according to their mitochondrial cytochrome b and nuclear rhodopsin gene sequences. The method was successfully tested in more than 200 marine fish species comprising the main Actinopterygii family groups. When used in phylogenetic analyses, its combination of two genes with different evolutionary rates serves to identify fish at the species level. We provide a flow diagram indicating our validated polymerase chain reaction amplification conditions for barcoding and species identification applications as well as population structure or haplotyping analyses, adaptable to high‐throughput analyses.     

Molecular phylogenetic relationship of Eplnephelus based on sequences of mtDNA Cty b

The mtDNA Cyt b gene was sequenced partially for Variola louti of Serranidae,Epinephelinae and seven endemic species of groupers-Epinephelus awoara,E.brunneus,E.coioides,E.longispinis,E.sexfasciatus,E.spilotoceps and E.tauvina in China.The seven endemic species and other seven foreign species of groupers--E,aeneus,E.caninus,E.drummondhayi,E,haifensis,E.labriformis,E.marginatus and E.multinotatus from the GenBank were combined and analysed as ingroup,while Variola louti was used as outgroup.We compared the 420 bp sequences of Cyt b among the 15 species and constructed two types of molecular phylogenetic trees with maximum parsimony method (MP)and neighbor-joining method (NJ) respectively.The results were as follows:(1) As to the base composition of mtDNA Cyt b sequence (402 bp) of 14 species of Epinepkelus,the content of (A + T) was 53.6%,higher than that of (G + C) (46.4%).The transition/transversion ratio was 4.78 with no mutation saturation.(2) The duster relationships between E.awoara and E.sexfasciatus,E.coioides and E.tauvina,E.longispinis and E.spilotoceps were consistent with phenotypes in taxonomy.(3) In the phylogenetic tree,the species in the Atlantic Ocean were associated closely with those in the Pacific Ocean,which suggested that the Cyt b sequences of Epinephelus were highly conserved.This may be attributed to the coordinate evolution.(4) In well-bred mating or heredity management,mating Epinephelus of the same branch should be avoided.It is likely to be an effective way to mate the species of the Atlantic Ocean with those of the Pacific Ocean to improve the inheritance species.     

从细胞色素b部分序列探讨石斑鱼属的分子系统发育关系

石斑鱼的分类在学术界尚存在很多争议。本文采用PCR技术,测定了分布于中国的点带石斑鱼(Epinepheluscoioiaes)、青石斑鱼(E.awoara)、六带石斑鱼(E.sexfasciatus)、褐石斑鱼(E.brunneus),长棘石斑鱼(E.longispinis),吻斑石斑鱼(E.spilotoceps),巨石斑鱼(E.tauvina)以及石斑鱼亚科侧牙鲈属的侧牙鲈(Variola louti)的线粒体细胞色素b基因402bp的序列,结合GenBank提供的7种中国无记录的石斑鱼:青铜石斑鱼(E.aeneus)、海丰石斑鱼(E.haifensis)、犬牙石斑鱼(E.caninus)、东大西洋石斑鱼(E.marginatus)、白斑石斑鱼(E.multinotatus)、德氏石斑鱼(E.drummondhayi)、淡点石斑鱼(E.labriformis),把这14种石斑鱼作为内群,对他们的序列组成和结构特点进行了分析;并以同为科(Serranidae)、石斑鱼亚科(Epinephelinae)的侧牙鲈属(Variola)的侧牙鲈作外群,分别用MP法、NJ法对内、外群构建了分子系统树,结果表明:(1)14种石斑鱼402bp的mtDNA Cytb部分序列的碱基组成上,A+T的含量为53.6%高于G+C含量(46.4%),序列中转换颠换比为4.78,没有突变饱和;(2)青石斑鱼与六带石斑鱼、点带石斑鱼与巨石斑鱼、长棘石斑鱼与吻斑石斑鱼分别聚在一起,与形态分类结果一致;(3)系统树中,大西洋与太平洋种类各枝交替汇合,表明石斑鱼的Cytb序列有严格的保守性,也可能是协同进化的结果;(4)揭示在良种选配或遗传管理时,避免居于同枝的石斑鱼混交,将大西洋种类与太平洋种类杂交可能是遗传改良的一种途径。       

Molecular phylogeny of three subspecies of common carp Cyprinus carpio, based on sequence analysis of cytochrome b and control region of mtDNA

The complete cytochrome b and the control region of mtDNA (about 2070 bp in total) of 10 strains belonging to three subspecies of the common carp, including three wild subspecies (the Yangtze River wild common carp – Cyprinus carpio haematopterus , Yuanjiang River wild common carp – Cyprinus carpio rubrofuscus and Volga River wild common carp – Cyprinus carpio carpio ) and seven domestic strains (Xingguo red carp, Russian scattered scaled mirror carp, Qingtian carp, Japanese Koi carp, purse red carp, Big-belly carp, German mirror carp) were sequenced. Phylogenetic analysis indicated that the 10 strains form three distinct clades, corresponding to C. c. haematopterus , C. c. rubrofuscus and C. c. carpio respectively. Purse red carp, an endemic domestic strain in Jiangxi province of China, showed a higher evolution rate in comparison with the other strains of C. c. haematopterus , most probably because of intensive selection and a long history of domestication. Base variation ratios among the three subspecies varied from 0.78% (between C. c. haematopterus and C. c. rubrofuscus ) to 1.47%(between C. c. carpio and C. c. rubrofuscus ). The topography of the phylogenetic tree and the geographic distribution of three subspecies closely resemble each other. The divergence time between C. c. carpio and the other two subspecies was estimated to be about 0.9 Myr and about 0.5 Myr between C. c. haematopterus and C. c. rubrofuscus . Based on phylogenetic analysis, C. c. rubrofuscus might have diverged from C. c. haematopterus .     

Mechanism of proton-pumping in the cytochrome b/f complex

Several models have been proposed to interpret the mechanism of proton-pumping associated with the electron transfer reactions in the cytochrome b/f complex. Energetics considerations suggest that the proton pump is coupled to the oxidation of cytochrome b by plastoquinone. Experiments performed in living cells under anaerobic conditions suggest that proton-pumping can occur through two independent mechanisms. When the two b cytochromes are reduced prior to a flash illumination i.e. after a long dark anaerobic incubation (>10 minutes), proton-pumping is very likely associated with the reduction of a semiquinone by cyt b which occurs at a site close to the inner face of the membrane. The electrogenic phase is associated with the tranfer of protons via a transmembrane channel. This process is not inhibited by 2-n-nonyl-4-hydroxyquinoline N-oxide (NQNO). Under repetitive-flash or under aerobic conditions, proton-pumping occurs according to a modified Q-cycle mechanism, which is inhibited by NQNO.Dedicated to Prof. L.N.M. Duysens on the occasion of his retirement