Relationships among orders and families of marsupials based on 12S ribosomal DNA sequences and the timing of the marsupial radiation

Part of the mitochondrial 12S ribosomal RNA gene was amplified and sequenced for 26 marsupials. Multiple alignments for these sequences as well as seven additional sequences taken from GenBank were obtained using CLUSTAL. PAUP was used for phylogenetic analysis and to obtain random tree-length distributions. Analyses were performed with and without phylogenetic constraints. Our results clearly show that 12S rDNA contains phylogenetic signal at and above the ordinal level and is thus appropriate for addressing phylogenetic questions deep in the mammalian tree. Standard parsimony analyses provide some support for a clade containing diprotodontians, dasyurids,Dromiciops, andNotoryctes; transversion parsimony analysis suggests the possible inclusion of peramelids as well. Within the Diprotodontia, vombatids and phascolarctids cluster together on transversion parsimony and phalangerids may be associated with this clade. The enigmatic tarsipedids are apparently part of a clade that also contains pseudocheirids, petaurids, and acrobatids. The 12S sequences suggest that the origination of extant marsupial orders peaked 15 million years later than the equivalent taxonomic diversification of extant placental orders and may be entirely post-Cretaceous. Families of diprotodontian marsupials originated during the Eocene and early Oligocene, which is consistent with previous single-copy DNA hybridization results.     

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 .     

Analysis of genetic diversity of domestic water buffaloes and anoas based on variations in the mitochondrial gene for cytochrome b

There are two major groups of domestic water buffaloes in East and Southeast Asia: swamp buffaloes and river buffaloes. Genetic diversity among swamp and river buffaloes was studied by DNA sequence analysis of the mitochondrial gene for cytochrome b. The results showed that each of the two groups has mitochondrial DNA (mtDNA) with a specific cytochrome b haplotype. The pairwise nucleotide sequence divergence was calculated to be 2·67% between swamp and river buffaloes, suggesting that they might have diverged from the ancestral populations of Asiatic domestic water buffaloes, approximately 1 million years ago. In addition, the sequences of the same gene from three subspecies of anoa (lowland, mountain and quarles anoa) were determined and compared with that of a domestic water buffalo. The sequence divergence was 1·2% for mountain anoa vs quarles anoa, 3·6% for mountain anoa vs lowland anoa and 3·3% for quarles anoa vs lowland anoa. Moreover, the sequence divergence between water buffaloes and anoas was found to be approximately 3·33%. Our results provide molecular evidence to support the taxonomic classification, namely, that Asiatic buffaloes may be classified into four lineages, swamp buffalo, river buffalo, lowland anoa and mountain plus quarles anoa. However, the sequence divergence values among these four groups were lower than the sequence divergence values found in the genus and subgenus levels within the subfamily Bovinae. In particular, in contrast to some proposed taxonomic classifications, our results indicated that mtDNA in the water buffaloes and anoas did not diverge at the genus level.     

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.     

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.     

Patterns of genetic variation in Pacific island land snails: the distribution of cytochrome b lineages among Society Island Partula

The radiation of Partula land snails has produced a large array of distinct morphological, ecological and behavioural types occupying many tropical volcanic islands in the Pacific Ocean. Within the Society Islands of French Polynesia, the mode of evolution is thought to have involved a single colonization event on each island, with later speciation occurring largely in situ. The present study examines genetic variation in the mitochondrial cytochrome b gene among taxa within the Society Island archipelago. Levels of intraspecific variation are found to be high, but variation among species is sometimes small. Mitochondrial variants do not always cluster according to island and some species are found to be polyphyletic in the cytochrome b tree, despite other morphological and molecular evidence that strongly supports their monophyly. A possible explanation for the polyphyly of species is that different variants are derived from ancestral mitochondrial polymorphisms that have been retained despite speciation events. Although it is possible that there has been some gene flow among islands, the distribution of mitochondrial lineages across islands strongly indicates that their origins predate the colonization of the islands in the study, and that they are very unlikely to have evolved entirely in situ.     

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。     

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。     

Evolution of trophic types in emperor fishes ( Lethrinus,Lethrinidae, Percoidei) based on cytochrome B gene sequence variation

Three trophic categories exist within emperor fishes, genus Lethrinus, relating to body form and dentition type. One group contains low-bodied, high speed, stalking predators with conical teeth. Another group comprises high-bodied, slow speed carnivores with molariform teeth capable of crushing hard-shelled benthic prey. A third group is also high-bodied but with conical teeth feeding mostly on small or soft-shelled benthic prey. Inferring the evolution of these trophic types within Lethrinus using morphology is problematic since these characters are typically correlated with feeding mode and are potentially homoplasious. We use mitochondrial DNA sequences, to independently determine a phylogenetic hypothesis for Lethrinus, which are not dependent on morphological characters relating to trophic categories. We analyzed complete cytochrome b gene sequences (1140 bp) for 20 species of Lethrinus, representing the three trophic types, and for 13 outgroup species, including four other representatives of the Lethrinidae. A monophyletic Lethrinidae did not resolve, but the monophyly of Lethrinus is well supported. In addition, two major clades within Lethrinus are well supported. One of these clades exclusively contains low-bodied species with conical teeth while the other clade only comprises the high-bodied species with molariform teeth. A high-bodied species with conical teeth, Lethrinus miniatus, appears most ancestral and sister to all other Lethrinus species. We hypothesize that this generalist trophic type was the evolutionary precursor to both of the other primary trophic types.     

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 among four species and a sub-species of Mullidae (Actinopterygii; Perciformes) based on mitochondrial cytochrome B, 12S rRNA and cytochrome oxidase II genes

DNA sequence comparisons of three mitochondrial DNA genes were used to reveal phylogenetic relationships among four species and a sub-species of Mullidae family. This is the first report using mitochondrial DNA sequence data to infer intraspecific relationship among different populations of Mullus barbatus and Mullus surmuletus; phylogenetic relationships between M. barbatus and its sub-species; M. barbatus ponticus. Cytochrome b, 12S ribosomal RNA, and cytochrome oxidase II regions of 242 individuals belonging to species M. barbatus, M. surmuletus, Upeneus moluccensis, Upeneus pori and sub-species M. barbatus ponticus were sequenced and phylogenetic trees were constructed using four different algorithms. The phylogenetic trees constructed support the existing taxonomical data of two mullid genera (Mullus, Upeneus). Molecular data shows no significant difference between same species of different geographical populations. The results suggest that the molecular difference is not large enough between M. barbatus and M. barbatus ponticus to consider them as sub-species.     

The largemouth bass cytochrome b gene

The cytochrome b gene is one of the protein-coding genes of the mitochondrial genome that has gained importance because of the ease with which molecular techniques can be applied to the analysis of its structure. The nucleotide sequence of the largemouth bass ( Microplerus salmoides ) cytochrome b gene was determined and the inferred amino acid structure is presented in the form of a structural model derived originally from rat cytochrome b . The inferred amino acid sequences from divergent animal species are aligned and compared in the context of this model. The data suggest that regions of the gene may be evolving at different rates due to different selection pressures associated with functional constraints. Conserved and variable regions of cytochrome b have been identified and can be targeted for species identification, the examination of intraspeciflc variation, and phylogenetic reconstructions in future research.     

Molecular phylogenetic relationships of China Seas groupers based on cytochrome <Emphasis Type="Italic">b</Emphasis> 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 T _S/T _V 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.     

Genetic diversity of mitochondrial cytochrome b gene in Chinese native buffalo

The origins of the domestic water buffalo remain contentious. To better understand the origins of Chinese water buffalo, we sequenced the complete mitochondrial cytochrome b (MT-CYB) gene from 270 individuals representing 13 Chinese domestic swamp buffalo populations. We found genetic evidence of introgression of river buffalo into Chinese swamp buffalo herds. Swamp buffalo haplotypes can be divided into two highly divergent lineages (A and B), suggesting that Chinese native swamp buffalo have two maternal origins. We found that the A→G transition in the buffalo MT-CYB gene stop codon resulted in buffalo haplotypes being terminated by one of two stop codons: AGA or AGG. AGA is common to river buffalo and lineage A of swamp buffalo, while AGG is specific to lineage B of swamp buffalo. Lineage A appears to have been domesticated in China. Further genetic evidence is required to clarify the origins of lineage B.     

Novel primers for complete mitochondrial cytochrome b gene sequencing in mammals

Sequence-based species identification relies on the extent and integrity of sequence data available in online databases such as GenBank. When identifying species from a sample of unknown origin, partial DNA sequences obtained from the sample are aligned against existing sequences in databases. When the sequence from the matching species is not present in the database, high-scoring alignments with closely related sequences might produce unreliable results on species identity. For species identification in mammals, the cytochrome b (cyt b) gene has been identified to be highly informative; thus, large amounts of reference sequence data from the cyt b gene are much needed. To enhance availability of cyt b gene sequence data on a large number of mammalian species in GenBank and other such publicly accessible online databases, we identified a primer pair for complete cyt b gene sequencing in mammals. Using this primer pair, we successfully PCR amplified and sequenced the complete cyt b gene from 40 of 44 mammalian species representing 10 orders of mammals. We submitted 40 complete, correctly annotated, cyt b protein coding sequences to GenBank. To our knowledge, this is the first single primer pair to amplify the complete cyt b gene in a broad range of mammalian species. This primer pair can be used for the addition of new cyt b gene sequences and to enhance data available on species represented in GenBank. The availability of novel and complete gene sequences as high-quality reference data can improve the reliability of sequence-based species identification.     

Phylogenetic Relationships and the Radiation of Sigmodontine Rodents in South America: Evidence from Cytochrome b

Phylogenetic relationships among South American sigmodontine rodents were examined based on the complete sequence for the mitochondrial cytochrome b gene [1140 base pairs (bp)] for 66 species and between 759 and 1140 bp for an additional 19 species. Thirty-eight South American genera were represented, coming from eight of nine tribes. Outgroups included the North American murid rodents Peromyscus, Reithrodontomys, Scotinomys, and Neotoma, the Old World murine rodents Mus and Rattus, and the geomyoid genera Thomomys, Geomys, Dipodomys, and Perognathus as the most distant outgroup. The South American sigmodontines were supported as a monophyletic lineage. Within this radiation several clear-cut suprageneric groupings were identified. Many of the currently recognized tribal groupings of genera were found fairly consistently, although not always with high levels of bootstrap support. The various tribes could not be linked hierarchically with any confidence. In addition, several genera stand out as unique entities, without any apparent close relatives. The overall pattern suggests a rapid radiation of the sigmodontines in South America, followed by differentiation at the tribal and generic levels.     

Relationships among several species of the subfamily Gymnelinae (Zoarcidae) inferred from mitochondrial DNA sequence analysis

Nucleotide sequences of the mtDNA COI and cytochrome b genes were determined in Magadanichthys skopetsi, a member of the new monotypic genus Magadanichthys, endemic to the northern coast of the Sea of Okhotsk. Comparison of this species with other representatives of the subfamily Gymnelinae (family Zoarcidae) revealed high genetic similarity of M. skopetsi to Hadropareia middendorffii and considerable differences between these species and Gymnelopsis ochotensis.