Investigations into the perplexing interrelationship of the Genus Takifugu Abe, 1949 (Tetraodontiformes, Tetraodontidae)

The phylogenetic relationships within the genus Takifugu Abe, 1949 (Tetraodontiformes, Tetraodonti- dae) remain unresolved. Because of the use of Takifugu as model organisms, the resolution of these relationships is crucial for the interpretation of evolutionary trends in biology. Pufferfishes of this ge- nus are comprised of a comparatively small number of species and are mainly distributed along the coastal region of the western part of the Sea of Japan and the coastline of China. Mitochondrial gene sequences were employed to test the phylogenetic hypotheses within the genus. Seventeen species of the genus were examined. Molecular phylogenetic trees were constructed using the maximum parsi- mony, neighbor-joining, maximum likelihood and Bayesian methods. Our hypothesis of internal relationships within the genus differs from previous hypotheses. Our results indicate that (1) the genus Takifugu is a monophyletic assemblage; (2) the genus is divided into 6 subgroups based on the mo- lecular data; and (3) there is low genetic diversity among the species within this genus. In addition, speciation within Takifugu appears to be driven by hybridization and isolation by distribution. Our re- sults also suggested that the taxonomy in the genus should be clarified based on both molecular and morphological data.     

Bayesian mixed models and divergence time estimation of Chinese cavefishes （Cyprinidae： Sinocyclocheilus）

The genus Sinocyclocheilus is distributed in Yun-Gui Plateau and its surrounding region only, within more than 10 cave species showing different degrees of degeneration of eyes and pigmentaUon with wonderful adaptations. To present, published morphological and molecular phylogenetic hypotheses of Slnocyclocheilus from prior works are very different and the relationships within the genus are still far from clear. We obtained the sequences of cytochrome b （cyt b） and NADH dehydrogenase subunlt 4 （ND4） of 34 species within Sinocyclocheilus, which represent the most dense taxon sampling to date. We performed Bayesian mixed models analyses with this data set. Under this phylogenetic framework, we estimated the divergence times of recovered clades using different methods under relaxed molecular clock. Our phyloegentic results supported the monophyly of Sinocyclocheilus and showed that this genus could be subdivided into 6 major clades. In addition, an earlier finding demonstrating the polyphyletlc of cave species and the most basal position of S. jii was corroborated. Relaxed divergence-time estimation suggested that Sinocyclocheilus originated at the late Miocene, about 11 million years ago （Ma）, which is older than what have been assumed.     

Bayesian mixed models and divergence time estimation of Chinese cavefishes (Cyprinidae: Sinocyclocheilus)

The genus Sinocyclocheilus is distributed in Yun-Gui Plateau and its surrounding region only, within more than 10 cave species showing different degrees of degeneration of eyes and pigmentation with wonderful adaptations. To present, published morphological and molecular phylogenetic hypotheses of Sinocyclocheilus from prior works are very different and the relationships within the genus are still far from clear. We obtained the sequences of cytochrome b (cyt b) and NADH dehydrogenase subunit 4 (ND4) of 34 species within Sinocyclocheilus, which represent the most dense taxon sampling to date. We performed Bayesian mixed models analyses with this data set. Under this phylogenetic framework, we estimated the divergence times of recovered clades using different methods under relaxed mo-lecular clock. Our phyloegentic results supported the monophyly of Sinocyclocheilus and showed that this genus could be subdivided into 6 major clades. In addition, an earlier finding demonstrating the polyphyletic of cave species and the most basal position of S. jii was corroborated. Relaxed diver-gence-time estimation suggested that Sinocyclocheilus originated at the late Miocene, about 11 million years ago (Ma), which is older than what have been assumed.     

Phylogenetic position of three Condylostoma species （Protozoa,Ciliophora, Heterotrichea） inferred from the small subunit rRNA gene sequence

The systematically poorly known ciliate genus Condylostoma was erected by Vincent in 1826. About 10 morphotypes have been reported, but any molecular investigations concerning this group so far are lacking. In this work, the small subunit ribosomal RNA （SS rRNA） gene of three marine Condylostoma species was sequenced, by which the phylogenetic trees were constructed by distancematrix, maximum parsimony and Bayesian inference methods. The results show that （1） all the trees have similar topologies with high supports; （2） Condylostoma is mostly related to the genus Condylostentor; and （3） three Condylostoma species as well as Condylostentor auriculatus cluster together and form a sister group with other heterotrichs. This is moderately consistent with the assessment of phylogenetic relationships of Condylostoma-related heterotrichs from the morphological information. The phylogenetic relationship of some other related heterotrichs, Peritromus, Folliculina, Stentor and Blepharisma, has been also discussed.     

Phylogenetic relationship and morphological evolution in the subfamily Limenitidinae （Lepidoptera： Nymphalidae）

The mitochondrial cytochrome oxidase subunit I （CO1） gene and the nuclear elongation factor 1 α（EF-1α） gene were sequenced from 29 species of Nymphalidae （Nymphalidae, Lepidoptera）. Phylogenetic trees were constructed based on the sequences determined from the 29 species and sequences of other 36 species deposited in GenBank using the neighbor-joining （NJ）, maximum likelihood （ML） and Bayesian methods with Libythea celtis （Libytheinae） as the outgroup. Our phylogenetic trees indicated four major clades. Clade A includes three subfamilies： Apaturinae, Nymphalinae, and Limenitidinae, excluding the tribe Limenitidini; Clade B includes the subfamilies Heliconiinae and the tribe Limenitidini; Clade C includes Satyrinae, Calinaginae, Charaxinae and Morphinae; and Clade D includes subfamily Danainae. Our study suggested that the tribes Pseudergolini, Biblidini, Limenitidini and Cyrestidini should be considered as subfamilies and confirmed the interspecific relationships within the subfamily Pseudergolinae, namely Amnosia ＋（Pseudergolis ＋ （Stibochiona ＋ Dichorragia））. We then mapped three morphological characters （spot of anal angle, eyespots, and process from outer margin of hind wing） onto the phylogenetic tree constructed by ML analysis using the combined sequence data. Based on this the evolutionary patterns of these morphological characters were identified, they indicated that the three characters evolved repeatedly in the family Nymphalidae.     

Phylogenetic position of Ephedra rhytidosperma, a species endemic to China： Evidence from chloroplast and ribosomal DNA sequences

The chloroplast genes matK and rbcL, ribosomal gene 18S and ITS regions of nuclear ribosomal DNA from Ephedra rhytidosperma, a species endemic to China, were sequenced and its phylogenetic position was investigated. Independent and combined phylogenetic analyses for the DNA sequences from 16 taxa representing 15 species of the genus Ephedra were performed using the maximum parsimony （MP）, neighbor-joining （N J）, minimum evolution （ME） and maximum likelihood （ML） methods. The results indicate that E. rhytidosperma is closely related to E. equisetina. The divergence time between them is estimated to be 10.85±2.44 Ma based on the results of the relative-rate tests and the evolutionary rate of rbcL gene.     

Phylogenetic relationships and divergence times of the family Araucariaceae based on the DNA sequences of eight genes

Araucariaceae is one of the most primitive families of the living conifers, and its phylogenetic relationships and divergence times are critically important issues. The DNA sequences of 8 genes, i.e., nuclear ribosomal 18S and 26S rRNA, chloroplast 16S rRNA, rbcL, matK and rps4, and mitochondrial coxl and atpl, obtained from this study and GenBank were used for constructing the molecular phylogenetic trees of Araucariaceae, indicating that the phyiogenetic relationships among the three genera of this family should be （（Wollemia, Agathis）, Araucaria）. On the basis of the fossil calibrations of Wollemia and the two tribes Araucaria and Eutacta of the genus Araucaria, the divergence time of Araucariaceae was estimated to be （308± 53） million years ago, that is, the origin of the family was in the Late Carboniferous rather than Triassic as a traditional view. With the same gene combination, the diver- gence times of the genera Araucaria and Agathis were （246 ± 47） and （61 ±15） Ma, respectively. Statis- tical analyses on the phylogenetic trees generated by using different genes and comparisons of the divergence times estimated by using those genes suggested that the chloroplast matK and rps4 genes are most suitable for investigating the phylogenetic relationships and divergence times of the family Araucariaceae.     

Molecular phylogeny and biogeography of the highly specialized grade schizothoracine fishes （Teleostei： Cyprinidae） inferred from cytochrome b sequences

We recovered the phylogenetic relationships among 23 species and subspecies of the highly special-ized grade schizothoracine fishes distributing at 36 geographical sites in the Tibetan Plateau and its surrounding regions by analyzing sequences of cytochrome b genes. Furthermore, we estimated the possible divergent times among lineages based on a historical geological isolation event in the Tibetan Plateau. The molecular data revealed that the highly specialized grade schizothoracine fishes were not a monophyletic group, but were the same as genera Gymnocypris and Schizogypsis. Our results indi-cated that the molecular phylogenetic relationships apparently reflected their geographical and historical associations with drainages, namely species from the same and adjacent drainages clustered together and had close relationships. The divergence times of different lineages were well consistent with the rapid uplift phases of the Tibetan Plateau in the late Cenozoic, suggesting that the origin and evolution of schizothoracine fishes were strongly influenced by environment changes resulting from the upheaval of the Tibetan Plateau.     

The phylogenetic relationship of the family Lutjanidae based on analyses of AFLP and mitochondrial 12S rRNA sequences

Fishes of the family Lutjanidae are commercially important in South China Sea. However, the phylogeny of Lutjanids is still unclear and there are many controversies over it. Herein, studies about the phylogeny of Lutjanids were performed based on Amplified Fragment Length Polymorphism （AFLP） analysis of genome DNA and sequence analysis of mitochondrial 12S rRNA gene, and 10 Lutjanidae species and 1 Lethrinidae species were employed. The topologies of minimum evolution （ME） trees based on the two analyses respectively were congruent except for positions of genera Pristipomoides and Caesio. The optimal substitution model TrN ＋ G for DNA sequences of 12S rRNA genes in Lutjanids was obtained using MODELTEST 3.6 software and maximum likelihood （ML） analysis supports the topology displayed by the ME tree. The test of log-likelihood suggests that the use of molecular clock calibrations to estimate species divergence time appeared valid. Phylogenetic analyses using AFLP data and DNA sequences of mitochondrial 12S rRNA genes indicated the monophyly of Lutjanus genra. However, further studies are required to reveal the phylogenetic relationship among other genera. In addition, the results demonstrated that AFLP genetic marker was suitable for the phylogenetic analysis of Lutjanids.     

Mitochondrial 16S rRNA sequence variations and phylogeny of the Chinese sisorid catfishes

Partial sequences of mitochondrial 16S rRNA gene were obtained by PCR amplification for comparisons among nine species of glyptosternoid fishes and six species of non-glyptosternoids representing 10 sisorid genera. There are compositional biases in the A-rich unpaired regions and G-rich paired regions. A-G transitions are primarily responsible for the Ts/Tv bias in unpaired regions. The overall substitution rate in unpaired regions is almost two times higher than that in the paired regions. Saturation plots at comparable levels of sequence divergence demonstrate no saturation effects. Phylogenetic analyses using both maximum likelihood and Bayesian methods support the monophyly of Sisoridae. Chinese sisorid catfishes are composed of two majorlineages, one represented by (Gagata (Bagarius, Glyptothorax)) and the other by “glyptosternoids Pseudecheneis“. The glyptosternoids may not be a monophyletic group. A previous hypothesis referring to Pseudecheneis as the sister group of monophyletic glyptosternoids, based on morphological evidence, is not supported by the molecular data. Pseudecheneis is shown to be a sister taxon of Glaridoglanis. Pareuchiloglanis might be paraphyletic with Pseudexostoma and Euchiloglanis. Our results also support the hypothesis that Pareuchiloglanis anteanalis might be considered as the synonyms of Pareuchiloglanis sinensis, and genus Euchiloglanis might have only one valid species, Euchiloglanis davidi.     

Asian origin for Polystichum （Dryopteridaceae） based on rbcL sequences

Chloroplast rbcL sequences of 60 species of Polystichum sensu lato (s.l.), including 23 new sequences from southwest China, were used to assess the phylogenetic relationships within the genus. On the basis of estimated evolution rate of rbcL gene and the genetic distance data that passed relative-rate tests, we further estimated the divergence times between some clades of the genus. The phylogenetic relationships were inferred using the neighbor-joining and maximum-parsimony methods, both methods producing trees with completely congruent topology. These trees reveal that all species of Polystichum s.l. in this study (including Cyrtomium and Cyrtomidictyum) form a monophyletic group. The basal split in Polystichum s.l. separates a clade with all Asian members from a clade containing other species from all over the world. The phylogenetic and divergence time estimation results lead us to suggest that Polystichum s.l. originated in Asia in the late Late Cretacous (≈76 Ma) and migrated into other places in the world in early Eocene(≈46 Ma).     

Morphological evolution from aquatic to terrestrial in the genus Oreolalax （Amphibia, Anura, Megophryidae）

A phylogeny of 17 species in the genus Oreolalax is reconstructed based on 21 morphological characters from adult specimens, skeleton specimens, tadpoles and eggs. Four species groups are recognized, of which the O. rugosus species group is the most primitive, the O. weigoldi species group is the second, the O. omeimontis species group is the third and the O. pingii species group is the most recently diversified. Based on the evolutional tendency of the morphological characters on the phylogenetic tree, it is proposed that the evolution of tympanum, tympanic annulus, columella, spoon-like cartilage and the web between toes reflect the habit changes from aquatic to terrestrial. Thus, Oreolalax is regarded as one important representative genus to study further the evolution of morphological characters from aquatic to terrestrial.     

Speciation of the genus Arthricocephalus Bergeron, 1899 (Trilobita) from the late Early Cambrian and its stratigraphic significance

The genus Arthricocephalus Bergeron, 1899 is revised, and Halipanktos Balker & Peel, 1997 is suggested here as a senior synonym. The subgenus Arthricocephalus (Arthricocephalites) Chien & Lin in Lu et al., 1974 is considered as a separate genus. Of the 20 previously assigned species of Arthricocephalus (Arthricocephalus) Bergeron, 1899, Arthricocephalus (Arthricocephalites) Chien & Lin in Lu et al., 1974, Arthricocephalus (Euarthricocephalus) Ju, 1983 are lumped into eight species. The speciation trend of Arthricocephalus and Arthricocephalites is demonstrated based on their stratigraphic occurrences. It not only enhances the resolution of the biostatigraphic zonation in the uppermost Lower Cambrian, but also represents a potential candidate to define the Duyunian stage. The base of the stage is suggested at the first appearance datum (FAD) of Arthricocephalus chauveaui Bergeron, 1899 within the evolutionary lineage from Ar. jiangkouensis Yin in Yin & Li,1978 to Ar. chauveaui in a global scale.     

Speciation of the genus Arthricocephalus Bergeron, 1899 (Trilobita) from the late Early Cambrian and its stratigraphic significance

The genus Arthricocephalus Bergeron, 1899 is revised, and Halipanktos Balker & Peel, 1997 is suggested here as a senior synonym. The subgenus Arthricocephalus (Arthricocephalites) Chien & Lin in Lu et al., 1974 is considered as a separate genus. Of the 20 previously assigned species of Arthricocephalus (Arthricocephalus) Bergeron, 1899, Arthricocephalus (Arthricocephalites) Chien & Lin in Lu et al., 1974, Arthricocephalus (Euarthricocephalus) Ju, 1983 are lumped into eight species. The speciation trend of Arthricocephalus and Arthricocephalites is demonstrated based on their stratigraphic occurrences. It not only enhances the resolution of the biostatigraphic zonation in the uppermost Lower Cambrian, but also represents a potential candidate to define the Duyunian stage. The base of the stage is suggested at the first appearance datum (FAD) of Arthricocephalus chauveaui Bergeron, 1899 within the evolutionary lineage from Ar. jiangkouensis Yin in Yin & Li,1978 to Ar. chauveaui in a global scale.     

中国白肩天蛾属记述与一新记录种(鳞翅目:天蛾科)

白肩天蛾属(Rhagastis RothschildJordan,1903)在中国分布有9种.于2017年上半年和2018年上半年分别对贵州荔波县茂兰自然保护区和海南乐东县尖峰岭地区进行考察并采集到一些天蛾标本.在对长喙天蛾族(Macroglossini Harris,1839)的标本进行整理之后发现,其中一些标本为宽缘白肩天蛾(Rhagastis acuta Walker,1856),这是该种首次在中国境内被记录.对该新记录种雄性成虫的形态进行了描述且附图,并对中国白肩天蛾属目前已知的种类进行了记述.     

New fossil elaterids （Insect： Coleoptera： Polyphaga： Elateridae） from the Middle Jurassic of Inner Mongolia, China

A new genus with a new species of fossil elaterid, Paraprotagrypnus superbus gen. et sp. nov., from the Middle Jurassic Jiulongshan Formation in Daohugou Village (N41°18.979′, E119°14.318′), Shantou Township, Ningcheng County, Inner Mongolia, China is described and illustrated. The genus Paraprotagrypnus belongs to the family of Elateridae, subfamily of Protagrypninae and tribe of Protagrypnini. Some primitive characters of the new genus and new species shed light on the ages between the Jiulongshan Formation in Inner Mongolia and the Yixian Formation in the western Liaoning Province of China. The habitat of the new species is briefly discussed.