A test of a mitochondrial gene-based phylogeny of woodpeckers (genus Picoides) using an independent nuclear gene, beta-fibrinogen intron 7

A conservative estimate of the species tree for the woodpecker genus Picoides based on two mitochondrial protein-coding genes is tested using sequences of an independently evolving nuclear intron, beta-fibrinogen intron 7. The mitochondrial gene-based topology and the intron-based topology are concordant, and a partition-homogeneity statistical test did not detect phylogenetic heterogeneity. The intron evolves more slowly than the mitochondrial sequences and tends not to resolve relationships among recently evolved species. However, the intron is superior over mitochondrial genes in resolving older bifurcations in the phylogeny. The two data sets were combined resulting in a robust estimate of the Picoides species tree in which most every node is statistically supported by bootstrap proportions. The Picoides species tree clearly shows that many morphological and behavioral characters used to lump species into this single genus have evolved by convergent evolution. Picoides is considered the largest genus of woodpeckers, but the molecular-based species tree suggests that Picoides is actually a conglomerate of several smaller groups.     

Molecular phylogeny of cockroaches (Blattaria) based on mitochondrial COII gene sequences

Phylogenetic relationships among five cockroach families (Cryptocercidae, Polyphagidae, Blattidae, Blattellidae and Blaberidae) using seventeen species, were estimated based on the DNA sequences of the mitochondrial cytochrome oxidase subunit II (COII) gene. A cladogram inferred using the neighbour‐joining method indicated that Polyphagidae and Cryptocercidae are closely related to each other, and that these two groups are a sister group to the remaining cockroach families. The monophyly of this clade, however, was not strongly supported in terms of bootstrap percentages. Blaberidae and Blattellidae were shown to be sister groups as previously proposed, with Blattidae as a sister group to that clade. Non‐weighted and weighted parsimony analyses were also performed following analyses of nucleotide substitution patterns that indicated saturation of the COII gene among these taxa had occurred. These parsimonious cladograms suggested that Polyphagidae was the basal family, and Polyphaginae, including Cryptocercus as proposed by Grandcolas 1994a ), was not monophyletic. The inferred relationships among cockroach families (Polyphagidae, Cryptocercidae + (Blattidae + (Blattellidae + Blaberidae))) is partly in agreement with some previously published analyses. Additionally, based on molecular data, Asian and American Cryptocercus are suggested to have diverged from one another before the Oligocene (～20 mya).     

基于线粒体基因cyt b的鹟亚科部分鸟类系统发育

采用分子系统学方法对鹟亚科(Muscicapinae)6属31种鸟类的cytb基因序列992bp进行系统发生分析。以荒漠伯劳(Lanius isabellinus)和发冠卷尾(Dicrurus hottentottus)为外群,采用贝叶斯法(Bayesian,BI)、最大似然法(Maximum-likelihood,ML)和最大简约法(Maximumparsimony,MP)分别构建鹟亚科的系统发育树。结果支持:寿带属(Terpsiphone)、扇尾鹟属(Rhipidura)与方尾鹟属(Culicicapa)可从鹟亚科中移出,其中寿带属归入王鹟科(Monarchidae),扇尾鹟属与方尾属归入扇尾鹟科(Rhipiduridae);鹟属(Muscicapa)、仙鹟属(Niltava)为单系发生,并聚为姐妹群,亲缘关系较近;姬鹟属(Ficedula)并非单系发生,白眉姬鹟(Ficedulazanthopygia)在3种系统发生树中的位置差别较大,研究结果未能确定其分类地位;铜蓝(Muscicapa thalassina)与白腹蓝(Cyanoptila cyanomelana)亲缘关系较近,前者应从属中移出,后者应从姬属移出,共同归入仙属或列为仙属的姐妹属。上述结论解决了亚科部分有争议属、种间的进化关系,为亚科分类系统提供了DNA水平证据。     

Molecular phylogeny of the genus Buteo (Aves: Accipitridae) based on mitochondrial marker sequences

DNA sequences of the mitochondrial nd6 gene and the non-repetitive part of the pseudo-control region (PsiCR) were isolated from 101 individuals to analyze the phylogenetic relationships among all buzzards of the genus Buteo and other buteonine genera. Comparisons of the two marker sequences indicate that the PsiCR evolved two times faster than the nd6 gene. The PsiCR proved to be an efficient, neutral genetic marker sequence for phylogenetic analyses at the intrageneric level, especially suitable for analyses based on old tissues, where only short fragments can be obtained. The molecular data set implies a neotropical origin of the genus Buteo. Monophyly of the genus Buteo as currently defined is contradicted due to the positions of Asturina nitida, Geranoaetus melanoleucus, Buteo magnirostris, and Buteo leucorrhous. These findings suggest several taxonomic consequences. A. nitida and G. melanoleucus should be included into the genus Buteo. Moreover, B. leucorrhous should be transferred into the genus Percnohierax (which clusters with Parabuteo), and B. magnirostris into the genus Rupornis. According to this classification of the genus Buteo, the basal lineage of the genus is formed by a clade containing Buteo polyosoma, Buteo poecilochrous, and Buteo melanoleucus. The "woodland buteos" form a paraphyletic assemblage with B. magnirostris as a clearly separated lineage basal to the genus Buteo.     

五种色蟌Cyt b基因序列及其系统发育关系

以昆虫mtDNA的Cytb基因作为分子遗传标记,用其特异性引物进行PCR扩增及DNA测序,共获得蜻蜓目束翅亚目色蟌科4属5种及1外群的Cytb基因部分序列(576bp),该片段中碱基T,C,A,G的平均含量分别为31.6,21.6,31.4和15.4%,A+T平均含量为63%,明显高于G+C含量(37%)。密码子第3位点的A+T平均含量较高,为66.4%。碱基替换多发生在密码子的第3位点。以巨齿尾溪蟌Bayadera melanopteryx作为外群构建系统发育树,结果显示,绿色蟌属Mnais和单脉色蟌属Matrona是单系群,绿色蟌属是单脉色属、细色蟌属Vestalis及艳色蟌属Neurobasis的姐妹群,是较早分化出来的一个类群。     

DNA barcoding of genus Toxoptera Koch (Hemiptera: Aphididae): Identification and molecular phylogeny inferred from mitochondrial COI sequences

Abstract Identification of aphid species is always difficult due to the shortage of easily distinguishable morphological characters. Aphid genus Toxoptera consists of species with similar morphology and similar to Aphis in most morphological characters except the stridulatory apparatus. DNA barcodes with 1 145 bp sequences of partial mitochondrial cytochrome‐coxidase I (COI) genes were used for accurate identification of Toxoptera. Results indicated mean intraspecific sequence divergences were 1.33%, whereas mean interspecific divergences were greater at 8.29% (0.13% and 7.79% if T. aurantii 3 and T. aurantii 4 are cryptic species). Sixteen samples were distinguished to four species correctly by COI barcodes, which implied that DNA barcoding was successful in discrimination of aphid species with similar morphology. Phylogenetic relationships among species of this genus were tested based on this portion of COI sequences. Four species of Toxoptera assembled a clade with low support in maximum‐parsimony (MP) analysis, maximum‐likelihood (ML) analysis and Bayesian phylogenetic trees, the genus Toxoptera was not monophyletic, and there were two sister groups, such as T. citricidus and T. victoriae, and two clades of T. aurantii which probably presented cryptic species in the genus.     

Molecular identification of Adoxophyes honmai (Yasuda) (Lepidoptera: Tortricidae) based on mitochondrial COI gene sequences

Molecular identification techniques are used where morphological characters are not useful for distinguishing species that resemble each other closely. The example studied here is the Adoxophyes species complex, in which A. orana (Fischer von R?sslerstamm) is officially the only known Korean species in the genus Adoxophyes (Lepidoptera: Tortricidae). However there have been suspicions that at least two types of A. orana exist in Korea based on the distribution and range of the host, with A. orana attacking apples and peaches, and another Adoxophyes sp. attacking tea and pears. The latter is presumed to be A. honmai (Yasuda), but the two have remained confused because of their extreme morphological similarity, despite several Asian studies of pheromonal and morphological characteristics. To confirm the occurrence of an Adoxophyes species other than A. orana in Korea, we compared 940 bp of the mitochondrial cytochrome oxidase I (COI) gene from 16 samples of Adoxophyes and found that there is a second Adoxophyes species different from A. orana. Comparison of the different sequences to that of Japanese A. honmai confirmed that they belong to the latter. From the sequence difference between the two Korean species, we were able to develop new PCR primer sets that distinguish them. This molecular identification technique with no enzyme digestion or sequencing step is a convenient and rapid way of differentiating between species that are hard to distinguish morphologically.     

Molecular phylogeny of the genus Oryzomys (Rodentia: Sigmodontinae) based on cytochrome b DNA sequences

The genus Oryzomys comprises 40 species arranged in several species groups. To test the monophyly of three Oryzomys species groups ("capito," nitidus, and subflavus), we analyzed, by distance, parsimony, and maximum-likelihood (ML), 801 bp of the mitochondrial gene cytochrome b. Our results did not sustain the monophyly of Oryzomys nor of the nitidus and subflavus species groups. Within the "capito" species group, O. perenensis appeared as a valid species, as a sister branch of the clade formed by O. megacephalus and O. laticeps. Within the nitidius species group, only the association between O. nitidus and O. lamia was well supported.The subflavus species group split into two clades: one with O. subflavus karyomorphotypes and another grouping O. angouya with species of different genera in the parsimony, distance, and ML trees.     

Molecular phylogeny of tephritid fruit flies in the Bactrocera tau complex using the mitochondrial COI sequences.

We compared sequences of the mitochondrial cytochrome oxidase I gene of eight species of the Bactrocera tau complex using Bactrocera dorsalis, Bactrocera pyrifoliae, Ceratitis capitata, Anopheles gambiae, and Locusta migratoria as outgroups. A 639-bp variable region was sequenced. The sequence divergence between species in the B. tau complex ranged from 0.06 to 28%, and up to 29% between the complex and its tephritid outgroups, B. dorsalis and C. capitata. According to the phylogenetic relationships, these members of the B. tau complex could be classified into four clades. Thus, species A and D form clades 1 and 3, respectively, while species C and I belong to clade 4. However, species B, E, and F form a distinct group, clade 2, and infested the fruits of non-cucurbit hosts. Host-plant shifts resulting in adaptive radiation and premating isolation among species might play an important role in species differentiation of the B. tau complex.     

Molecular phylogeny of the medaka fishes genus Oryzias (Beloniformes: Adrianichthyidae) based on nuclear and mitochondrial DNA sequences

The phylogenetic relationships among medaka fishes of 2 genera, Oryzias and Xenopoecilus, were studied using the nuclear tyrosinase and mitochondrial 12S and 16S rRNA genes. Of the 23 species currently described for these genera, 13 species of Oryzias and 2 species of Xenopoecilus were examined. The tree topologies obtained from the nuclear and mitochondrial data were consistent, indicating that Xenopoecilus is a polyphyletic genus nested within Oryzias. This result suggested the necessity for a systematic study and taxonomic revision of Xenopoecilus. The combined data analysis of all data partitions resulted in a well-resolved tree, with most internal branches supported by high statistical values. Based on our combined data phylogeny, we divided the Oryzias species into three major species groups, namely the latipes, javanicus, and celebensis groups. These three groups corresponded to the three chromosomal groups (biarmed, monoarmed, and fused chromosome groups) previously proposed from karyological analyses. The phylogeographic pattern suggests historical vicariance between Sulawesi Island and the continental shelf.     

A molecular phylogeny of the genus Gammarus (Crustacea: Amphipoda) based on mitochondrial and nuclear gene sequences

A phylogeny of the genus Gammarus Fabricius, 1775 was constructed using DNA sequence data from the mitochondrial genes COI and 16S, and the nuclear genes 18S and 28S. Both parsimony and Bayesian analyses were conducted on separate and combined data partitions. The Bayesian phylogeny from the combined analysis was selected as the preferred phylogenetic hypothesis. The hypothesis supports monophyly of the genus Gammarus, paraphyly of the European-North American Gammarus, and monophyly of the Asian Gammarus. The Asian clade was further split into a southeastern group and a northwestern group. The dramatic climate change following the uplift of the Tibetan Plateau was probably the most important factor in triggering the diversification of southeastern and northwestern groups. The genus Sinogammarus is invalid and should be part of the genus Gammarus.     

Molecular phylogeny of the speciose vole genus Microtus (Arvicolinae, Rodentia) inferred from mitochondrial DNA sequences

Voles of the genus Microtus represent one of the most speciose mammalian genera in the Holarctic. We established a molecular phylogeny for Microtus to resolve contentious issues of systematic relationships and evolutionary history in this genus. A total of 81 specimens representing ten Microtus species endemic to Europe as well as eight Eurasian, six Asian and one Holarctic species were sequenced for the entire cytochrome b gene (1140 bp). A further 25 sequences were retrieved from GenBank, providing data on an additional 23, mainly Nearctic, Microtus species. Phylogenetic analysis of these 48 species generated four well-supported monophyletic lineages. The genus Chionomys, snow voles, formed a distinct and well-supported lineage separate from the genus Microtus. The subgenus Microtus formed the strongest supported lineage with two sublineages displaying a close relationship between the arvalis species group (common voles) and the socialis species group (social voles). Monophyly of the Palearctic pitymyid voles, subgenus Terricola, was supported, and this subgenus was also subdivided into two monophyletic species groups. Together, these groupings clarify long-standing taxonomic uncertainties in Microtus. In addition, the Asian and the Nearctic lineages reported previously were identified although the latter group was not supported. However, relationships among the main Microtus branches were not resolved, suggesting a rapid and potentially simultaneous radiation of a widespread ancestor early in the history of the genus. This and subsequent radiations discernible in the cytochrome b phylogeny, show the considerable potential of Microtus for analysis of historical and ecological determinants of speciation in small mammals. It is evident that speciation is an ongoing process in the genus and that the molecular data provides a vital insight into current species limits as well as cladogenic events of the past.     

Molecular phylogeny of Fordini (Hemiptera: Aphididae: Pemphiginae) inferred from nuclear gene EF-1 alpha and mitochondrial gene COI

The tribe Fordini is a fascinating group because of its complicated life history, primary host specificity and gall-forming characteristic. Different species produce galls with different morphology on different parts of the host plants. The EF-1alpha-based, COI-based and combined sequences-based phylogenetic trees with three algorithms MP, ML and Bayes all strongly suggest that Fordini is a monophyletic group with two clades corresponding to two subtribes, Fordina and Melaphidina, each also monophyletic. Some important morphological characters and primary host plants of aphids were mapped onto the phylogenetic tree to analyse the division of subtribes and to uncover at which level the aphids correspond to their primary hosts, Pistacia and Rhus. Results suggest that the division of subtribes in Fordini is closely related to host selection of aphids. The evolution of gall morphology and the probable driving force behind it in this tribe were also discussed. The Fordini aphids seem to have evolved towards a better ability to manipulate their host plant, induce strong sinks and gain high reproductive success. Galls in this tribe evolved mainly along two directions to attain this goal: (i) by enlarging the gall from small bag to spherical, even big cauliflower-like, and changing the galls' location or forming two galls in their life cycle (Fordina); (ii) by moving the gall position from midrib, petiole of the leaflet, and eventually to the common petiole of the compound leaf (Melaphidina).     

Molecular phylogeny and biogeography of woolly flying squirrel (Rodentia: Sciuridae), inferred from mitochondrial cytochrome b gene sequences

To investigate the genetic diversity between the populations of woolly flying squirrels (Eupetaurus) from the eastern and western extremes of the Himalayas, partial mitochondrial cytochrome b gene sequences (390-810 bp) that were determined from the museum specimens were analyzed using maximum parsimony (MP) and maximum likelihood (ML) methods. The molecular data reveal that the two specimens that were collected in northwestern Yunnan (China) are members of the genus Eupetaurus. Reconstructed phylogenetic relationships show that the populations of Eupetaurus in the eastern and western extremes of the Himalayas are two distinct species with significant genetic differences (12%) and diverged about 10.8 million years ago. Eupetaurus is significantly different from Petaurista and Pteromys. The level of estimated pairwise-sequence divergence observed between Eupetaurus and Petaurista or Pteromys is greater than that observed between Eupetaurus and Trogopterus, Belomys, Glaucomys, or Hylopetes. Considering the divergence time of the two Eupetaurus groups, the glaciations and the uplift of the Himalayas and Qinghai-Tibet plateau during the Pliocene-Pleistocene period might be the major factors affecting the present distribution of Eupetaurus along the Himalayas.     

A molecular phylogeny of the marine mussel genus Perna (Bivalvia: Mytilidae) based on nuclear (ITS1&2) and mitochondrial (COI) DNA sequences

A molecular phylogeny is presented for marine mussels of the genus Perna, based on nuclear (ITS1,ITS2) and mitochondrial (COI) DNA sequence data. The three generally recognised species (Perna viridis, Perna perna and Perna canaliculus) and one putative species (Perna picta) were each sampled from several locations within their known geographic distributions. A range of phylogenetic analyses was used to investigate the current taxonomic assignments, evolutionary relationships and the biogeographical history of the genus. The different analyses produced similar, well supported topologies and verified the monophyly of the genus with respect to five mytilid outgroup species. P. perna (Atlantic), P. viridis (Indo-West Pacific), and P. canaliculus (New Zealand) each formed distinct clades, confirming their specific status. Putative P. picta from North Africa clustered within the P. perna clade and is not regarded as a separate species. P. perna and P. canaliculus were the most closely related of the three species. Possible biogeographic explanations for the present species distributions are evaluated.     

Phylogenetic and phylogeographic relationships in the crayfish genus Austropotamobius inferred from mitochondrial COI gene sequences

The present study explores the utility of mitochondrial COI gene sequences to reveal phylogenetic and phylogeographic relationships for the entire European freshwater crayfish genus Austropotamobius. The two traditional taxa, Austropotamobius pallipes and Austropotamobius torrentium, were monophyletic, showing similar genetic diversity, with 28 and 25 haplotypes, respectively, and an uncorrected average pairwise divergence of 0.059 and 0.041. A third distinct haplotype clade, in sister relation to A. torrentium, was discovered at the Upper Kolpa drainage in the northern Dinaric area. All populations north and west of the Alps are genetically impoverished (nucleotide diversity (pi)=0.000-0.001), while southern populations are more diverse (pi=0.001-0.034). A. pallipes reaches the highest diversity in the region of Istra, probably its primary center of radiation. The genetic diversity center for A. torrentium is the southern Balkan peninsula. Other potential glacial refugia were identified in Southern France, Northwestern Italy, the Apennine Peninsula, and in the northern Dinaric area. The Iberian Peninsula has been stocked artificially from Northern Italy. Three main periods of radiation were tentatively identified: late Miocene/early Pliocene for the divergence of species and main lineages, the Pleistocene for the divergence within populations south from Alps, and a postPleistocene expansion north and west from Alps.     

Molecular phylogeny and evolution of the freshwater eels genus Anguilla based on the whole mitochondrial genome sequences

Molecular phylogenetic analyses were conducted using the whole mitochondrial genome sequences of all 18 species/subspecies of the freshwater eels genus Anguilla to infer their phylogenetic relationships and to evaluate hypotheses about the possible dispersal routes of this genus. The Bayesian and maximum likelihood analyses using a total of 15,187 sites of mitochondrial DNA sequences suggested that A. mossambica was the most basal species of anguillid eel, and that the other species (except for A. borneensis) formed three geographic clades: Atlantic (two species), Oceania (three species), and Indo-Pacific (11 species). The present study clearly indicated a sister relationship between the Atlantic and Oceanian species, which now have distantly separated geographic distributions. This suggests that the previous hypotheses to estimate the dispersal route of anguillid eels into the Atlantic Ocean based on the current geographic distribution of species are unsupported by the present more complete analysis. Alternatively, the unique geographic distribution of the present day species in the genus Anguilla appears to have resulted from multiple dispersal events. Although the age of the beginning of speciation among anguillid eels was tentatively estimated as 20 million years ago using a calibration for bony fishes of 7.3x10(-4) substitutions/site/million years, it is possible that this divergence time was underestimated because of the ecological characteristics of these fishes. The results of the present study suggest that the hypotheses for the dispersal route and divergence time of the genus Anguilla should be reconsidered.     

Molecular phylogeny of Clupeiformes (Actinopterygii) inferred from nuclear and mitochondrial DNA sequences

The taxonomy of clupeiforms has been extensively studied, yet phylogenetic relationships among component taxa remain controversial or unresolved. Here we test current and new hypotheses of relationships among clupeiforms using mitochondrial rRNA genes (12S and 16S) and nuclear RAG1 and RAG2 sequences (total of 4749bp) for 37 clupeiform taxa representing all five extant families and all subfamilies of Clupeiformes, except Pristigasterinae, plus seven outgroups. Our results, based on maximum parsimony, maximum likelihood, and Bayesian analyses of these data, show that some traditional hypotheses are supported. These include the monophyly of the families Engraulidae, consisting of two monophyletic subfamilies, Engraulinae (Engraulis and Anchoa) and Coilinae (Coilia and Setipinna), and Pristigasteridae (here represented only by Ilisha and Pellona). The basal position of Denticeps among clupeiforms is consistent with the molecular data when base compositional biases are accounted for. However, the monophyly of Clupeidae was not supported. Some clupeids were more closely related to taxa assigned to Pristigasteridae and Chirocentridae (Chirocentrus). These results suggest that a major revision in the classification of clupeiform fishes may be necessary, but should await a more complete taxonomic sampling and additional data.