Phylogeny and evolution of Perezia （Asteraceae： Mutisieae： Nassauviinae）

A molecular phylogenetic analysis of most of the species of Perezia reveals that, as traditionally defined, the genus is not monophyletic with two species more closely related to Nassauvia than to Perezia. In addition, our results show that Burkartia （Perezia） lanigera is related to Acourtia and is the only member of that clade in South America. The remaining species are monophyletic and show a pattern of an early split between a western temperate and an eastern subtropical clade of species. Within the western clade, the phylogeny indicates a pattern of diversification that proceeded from southern, comparatively low-elevation habitats to southern high-elevation habitats, and ultimately into more northern high-elevation habitats. The most derived clades are found in the high central Andes, where significant radiation has occurred.     

Phylogeny of Ptychostomum （Bryaceae,Musci） inferred from sequences of nuclear ribosomal DNA internal transcribed spacer （ITS） and chloroplast rps4

The phylogeny of Ptychostomum was first spacer （ITS） region of the nuclear ribosomal （nr） DNA DNA rps4 sequences. Maximum parsimony, maximum undertaken based on analysis of the internal transcribed and by combining data from nrDNA ITS and chloroplast likelihood, and Bayesian analyses all support the conclusion that the reinstated genus Ptychostomum is not monophyletic. Ptychostomum funkii （Schwagr.） J. R. Spence （≡ Bryum funkii Schwaigr.） is placed within a clade containing the type species of Bryum, B. argenteum Hedw. The remaining members of Ptychostomum investigated in the present study constitute another well-supported clade. The results are congruent with previous molecular analyses. On the basis of phylogenetic evidence, we agree with transferring B. amblyodon Mull. Hal. （≡ B. inclinatum （Brid.） Turton≡ Bryum archangelicum Bruch ＆ Schimp.）, Bryum lonchocaulon Mull. Hal., Bryum pallescens Schleich. ex Schwaigr., and Bryum pallens Sw. to Ptychostomum.     

Systematic positions of Lamiophlomis and Paraphlomis （Lamiaceae） based on nuclear and chloroplast sequences

Genera Lamiophlomis and Paraphlomis were originally separated from genus Phlomis s.1. on the basis of particular morphological characteristics. However, their relationship was highly contentious, as evidenced by the literature. In the present paper, the systematic positions of Lamiophlomis, Paraphlomis, and their related genera were assessed based on nuclear internal transcribed spacer （ITS） and chloroplast rpl16 and trnL-F sequence data using maximum parsimony （MP） and Bayesian methods. and outgroup were sampled. Analyses of both separate In total, 24 species representing six genera of the ingroup and combined sequence data were conducted to resolve the systematic relationships of these genera. The results reveal that Lamiophlomis is nested within Phlomis sect. Phlomoides and its generic status is not supported. With the inclusion ofLamiophlomis rotata in sect. Phlomoides, sections Phlomis and Phlomoides of Phlomis were resolved as monophyletic. Paraphlomis was supported as an independent genus. However, the resolution of its monophyly conflicted between MP and Bayesian analyses, suggesting the need for expended sampling and further evidence.     

Phylogeny and Classification of Prunus sensu lato （Rosaceae）

The classification of the economically important genus Prunus L. sensu lato （s.L） is controversial due to the high levels of convergent or the parallel evolution of morphological characters. In the present study, phylogenetic analyses of fifteen main segregates of Prunus s.I. represented by eighty-four species were conducted with maximum parsimony and Bayesian approaches using twelve chloroplast regions （atpB- rbcL, matK, ndhF, psbA-trnH, rbcL, rpL 16, rpoC1, rps16, trnS-G, trnL, trnL-F and ycfl） and three nuclear genes （ITS, s6pdh and Sbel） to explore their infrageneric used to develop a new, phylogeny-based classification relationships. The results of these analyses were of Prunus s.I. Our phylogenetic reconstructions resolved three main clades of Prunus s.I. with strong supports. We adopted a broad-sensed genus, Prunus, and recognised three subgenera corresponding to the three main clades： subgenus Padus, subgenus Cerasus and subgenus Prunus. Seven sections of subgenus Prunus were recognised. The dwarf cherries, which were previously assigned to subgenus Cerasus, were included in this subgenus Prunus. One new section name, Prunus L. subgenus Prunus section Persicae （T. T. yu ＆ L. T. Lu） S. L. Zhou and one new species name, Prunus tianshanica （Pojarkov） S. Shi, were proposed.     

New combination in Veronica （Scrophulariaceae s.l.） based on morphological characters and the seed storage protein polymorphism

Veronica erciyasdagi （M. A. Fischer） C. Vural comb. ＆ stat. nov., previously regarded as a variety, is proposed as a new combination, based on the morphological characters and seed storage protein polymorphism presented in this study. In addition, information about the ecology and conservation status of Veronica erciyasdagi was reported. This taxon is endemic to central Anatolia, Turkey and is critically endangered.     

A New Species of the Genus Tylototriton （Urodela： Salamandridae） from Northeastern Hunan Province,China

We describe a new species from the family Salamandridae from northeastern Hunan Province of China based on morphological and molecular genetic evidences.The new species,Tylototriton liuyangensis,is a member of the T.asperrimus group,and shares a number of similarities with T.wenxianensis and T.broadoridgus.The dorsal surface of the new Tylototriton species is completely black,with nodule-like warts distributed evenly along the lateral margin of dorsal body,and there is no transverse striae between the warts.There are several diagnostic characters of the new species that differ from those of T.wenxianensis and T.broadoridgus as follows：（1）the interorbital space（IOS）is comparatively wide;（2）there is a bony ridge present on the dorsal head surface,running along the upper eyelids;（3）the space between axilla and groin（AGS）is comparatively large,with the mean male AGS approximately equal to 50%of the snout-vent length（SVL）and the mean female AGS being 53%of SVL;and（4）there is a small genital armature on the anterior angle of labial folds present inside the anal fissure of adult males during the breeding season.The result from the molecular phylogenetic analysis of the genus Tylototriton（including the type specimen of the new species）based on the complete DNA sequence of the mitochondrial ND2 gene（1035 bp）indicated that the new species was similar to T.wenxianensis,T.dabienicus,and T.broadoridgus,but formed an independent clade.This result was consistent with the morphological analysis above,supporting that the Liuyang Population represented a distinct species.     

Integrative taxonomy of Leptonetela spiders (Araneae,Leptonetidae), with descriptions of 46 new species

Extreme environments,such as subterranean habitats,are suspected to be responsible for morphologically inseparable cryptic or sibling species and can bias biodiversity assessment.A DNA barcode is a short,standardized DNA sequence used for taxonomic purposes and has the potential to lessen the challenges presented by a biotic inventory.Here,we investigate the diversity of the genus Leptonetela Kratochvíl,1978 that is endemic to karst systems in Eurasia using DNA barcoding.We analyzed 624 specimens using one mitochondrial gene fragment (COI).The results show that DNA barcoding is an efficient and rapid species identification method in this genus.DNA barcoding gap and automatic barcode gap discovery (ABGD) analyses indicated the existence of 90 species,a result consistent with previous taxonomic hypotheses,and supported the existence of extreme male pedipalpal tibial spine and median apophysis polymorphism in Leptonetela species,with direct implications for the taxonomy of the group and its diversity.Based on the molecular and morphological evidence,we delimit and diagnose 90 Leptonetela species,including the type species Leptonetela kanellisi (Deeleman-Reinhold,1971).Forty-six of them are previously undescribed.The female of Leptonetela zhai Wang & Li,2011 is reported for the first time.Leptonetela tianxinensis (Tong & Li,2008) comb.nov.is transferred from the genus Leptoneta Simon,1872;the genus Guineta Lin & Li,2010 syn.nov.is a junior synonym of Leptonetela;Leptonetela gigachela (Lin & Li,2010)comb.nov.is transferred from Guineta.The genus Sinoneta Lin & Li,2010 syn.nov.is a junior synonym of Leptonetela;Leptonetela notabilis (Lin & Li,2010) comb.nov.and Leptonetela sexdigiti (Lin & Li,2010) comb.nov.are transferred from Sinoneta;Leptonetela sanchahe Wang & Li nom.nov.is proposed as a replacement name for Sinoneta palmata (Chen et al.,2010) because Leptonetela palmata is preoccupied.     

Multilocus Phylogeny of Lycodon and the Taxonomic Revision of Oligodon multizonatum

Classification of the Asian snake genera Lycodon and Oligodon has proven challenging. We conducted a molecular phylogenetic analysis to estimate the phylogenetic relationships in the genus of Lycodon and clarify the taxonomic status of Oligodon multizonatum using mitochondrial（cyt b, ND4） and nuclear（c-mos） genes. Phylogenetic trees estimated using Maximum Likelihood and Bayesian Inference indicated that O. multizonatum is actually a species of Lycodon. Comparing morphological data from O. multizonatum and its closest relatives also supported this conclusion. Our results imply that a thorough review of the evolutionary relationships in the genus of Lycodon is strong suggested.     

Cytological study on the genus Syncalathium （Asteraceae-Lactuceae）, an endemic taxon to alpine scree of the Sino-Himalayas

Cytological characters of four species in Syncalathium （Asteraceae： Lactuceae）, a small genus with six identified species endemic to alpine scree of the Sino-Himalayan region, are surveyed in this report. Three species （Syncalathiumpilosum, Syncalathium chrysocephalum, and Syncalathium disciforme） are examined for the first time. Combined with our previous counts, five species have been cytologically investigated from the genus and the results indicated that all species are diploid with the basic somatic chromosome number of x=8. The karyotype asymmetry of Syncalathium souliei is 2A, distinct from the other four species of 1A, and the remaining species are divided into two subgroups with different karyotypes, consistent with their morphological features. The significance of the cytological evolution of Syncalathium is briefly discussed.     

A REVIEW OF THE GENUS A CRICOTOPUS KIEFFER （DIPTERA： CHIRONOMIDAE） IN CHINA

The genus Acricotopus Kieffer including three species from China is reviewed.Acricotopus lucens (Zetterstedt) is reexamined. Two species A. simplex sp. nov. and A.zhalingensis sp. nov. are described as male imagines. Acricotopus simplex sp. nov. was collected from Xizang, Sichuan and Qinghai Provinces; the species differs from others of the genus by the absence of hind tibial comb, temporal and clypeal seta. Acricotopus zhalingensis sp. nov. was collected from Qinghai Province; the species is recognized by a combination of characters: the larger body size; the clypeus without setae; anal lobe strongly produced, nearly hemispherical shape; inferior volsella triangular, strongly sclerotized; gonostylus with 13 very long setae. A key to the world males of the genus is presented. The generic diagnosis is emended.All the types described in this paper are deposited in Department of Biology, Nankai University,China (BDN).     

Phylogeny of genus Glossina (Diptera: Glossinidae) according to ITS2 sequences

The flies of genus Glossina (Diptera: Glossinidae) are an important vector of African trypanosomiases which cause diseases in humans and animals. The ribosomal DNA Internal Transcribed Spacer-2 (ITS-2) region sequences from different Glossina species were PCR-amplified and analyzed in order to construct a molecular phylogeny for genus Glossina. Trees generated by parsimony confirmed the monophyletic taxonomic placement of genus Glossina where fusca group species formed the deepest branch followed by morsitans and palpalis groups, respectively. The placement of Glossina austeni by both the traditional morphological and biochemical criteria has been controversial. Results presented here, based on ITS-2 locus sequence analysis, suggest that Glossina austeni can be placed into a separate subgenerus which forms a sister-group relationship with the morsitans group species.     

Phytogeny of 12 species of genus Glycine Willd. reconstructed with internal transcribed region in nuclear ribosomal DNA

The ITS-Is of 24 accessions belong to 10 species of subgenus Glycine, and 2 species of subgenus Soja of genus Glycine were amplified, cloned and sequenced. According to the homology of the sequences, the phy-logeny of the 24 accessions were reconstructed. The reconstructed dendrogram showed that there were some divergent genomic types found in the previously classified species, such as G . tomentella, G. canescens and G. tabacina, and they might be some cryptic species by morphologic analysis.     

Resolution of phylogenetic relationships of the major subfamilies of the Delphacidae （Homoptera： Fulgoroidea） using the mitochondrial ribosomal DNA

Delphacid relationships from the genus level to the subfamily have been completely resolved （among those taxa examined） using sequence data from the 3＇ end of the 12S gene. Monophyly of the non-asiracine subfamilies was strongly supported and the asiracine Ugyops was placed in the most basal position of the tree. Support levels for monophyly of the Delphacini increased after weighting transversions more heavily than transitions and after removing the cixiid outgroup from the dataset. Among the Delphacini, Conomelus and Megamelus were more closely related to each other than either was to Chloriona. These results are in agreement with the tree based on morphological characters. However, in contrast to morphological data our results strongly supported a sister group relationship between the Stenocraninae and the Kelisiinae. Although the 12S gene fragment gave some information about the species relationships within Chloriona, neither this fragment nor the 5＇ end of the 16S gene appear to be very useful for this level. Molecular evolutionary patterns provided evidence that there has been a shift in base composition from T to A during the early evolution of the non-Asiracinae. The non-Asiracinae also had comparatively fast substitution rates and these two observations are possibly correlated. In the ‘ modem＇ delphacid Chloriona, the AT content was comparatively low in regions free of constraints but this was not the case for ‘ non-modem＇ delphacids. The tRNA for valine has been translocated elsewhere, probably before the Delphacidae and Cixiidae diverged from each other.     

Phylogenetic relationships of Cyprinidae (Teleostei: Cypriniformes) inferred from the partial S6K1 gene sequences and implication of indel sites in intron 1

The family Cyprinidae is widely distributed in East Asia, and has the important phylogenetic signifi- cance in the fish evolution. In this study, the 5′ end partial sequences (containing exon 1, exon 2 and indel 1) of S6K1 gene were obtained from 30 representative species in Cyprinidae and outgroup using PCR amplification and sequencing. The phylogenetic relationships of Cyprinidae were reconstructed with neighbor joining (NJ), maximum parsimony (MP), maximum likelihood (ML), and Bayesian meth- ods. Myxocyprinus asiaticus (Catostomidae) was assigned to the outgroup taxon. Similar phylogenetic relationships within the family Cyprinidae were achieved with the four analyses. Leuciscini and Barbini were monophyletic lineages respectively with the high nodal supports. Leuciscini comprises Hy- pophthalmichthyinae, Xenocyprinae, Cultrinae, Gobioninae, Acheilognathinae and East Asian species of Leuciscinae and Danioninae. Monophyly of East Asian clade was supported with high nodal support. Barbini comprises Schizothoracinae, Barbinae, Cyprininae and Labeoninae. The monophyletic lineage consisting of Danio rerio, D. myersi, and Rasbora trilineata was basal in the tree. In addition, the large fragment indels in intron 1 were analyzed to improve the understanding of Cyprinidae relationships. The results showed that the large fragment indels were correlated with the relations among species. Some conserved regions in intron 1 were thought to be involved in the functional regulation. However, no correlation was found between sequence variations and species characteristic size.     

Geometric morphometric analysis of Eysarcoris guttiger,E.annamita and E.ventralis (Hemiptera: Pentatomidae)

The genus Eysarcoris can be easily distinguished from other genera through the two spots in the basal angle of the scutellum.Nevertheless,Eysarcoris species show complex variances.Geometric morphometric methods have been increasingly applied to distinguish species and to define the boundary of genera among insects.In the present study,geometric morphometric approach was firstly employed to evaluate the shape variation of three characters (fore wing,hind wing and pygophore) of E.guttiger,E.annamita and E.ventralis using E.aeneus as outgroup to ascertain whether this approach is a reliable method for the taxonomy of Eysarcoris.Analysis was conducted on the landmarks of the three characters of these species.Multivariate regression of procrustes coordinates against centroid size was conducted to test the presence of allometry.Principal component analysis (PCA),canonical variate analysis (CVA)and cluster analysis were utilized to describe variations in shapes among the studied species.For all of the three characters,though PCA analysis showed some overlap among species,p-values for procrustes distance and mahalanobis distance were all less than 0.0001.The distribution of the three studied species corresponds with their species status.This study demonstrates that the geometric morphometrics of both the fore wing and the hind wing might represent a possible tool for the identification of species within this genus.     

NOTES ON ANNOSA SPECIES-GROUP OF HELINA R.-D. （DIPTERA： MUSCIDAE） IN CHINA

The genus Helina R.-D., 1830 is the second largest genus in Muscidae. By now,more than 400 species are known in the world and about 200 species have been described in China. This paper outlines earlier studies on the subgenus and species-group of Helina. Helina annosa species-group is erected. A key to the known Chinese subgroups and species is given.The 19 known Chinese species is listed, including one new species, i.e. Helina setipostitibia sp. nov. Type specimen of the holotype of the new species is deposited in the Institute of Entomology, Shenyang Normal University.     

A fossil-calibrated relaxed clock for Ephedra indicates an Oligocene age for the divergence of Asian and New World clades and Miocene dispersal into South America

Ephedra comprises approximately 50 species, which are roughly equally distributed between the Old and New World deserts, but not in the intervening regions （amphitropical range）. Great heterogeneity in the substitution rates of Gnetales （Ephedra, Gnetum, and Welwitschia） has made it difficult to infer the ages of the major divergence events in Ephedra, such as the timing of the Beringian disjunction in the genus and the entry into South America. Here, we use data from as many Gnetales species and genes as available from GenBank and from a recent study to investigate the timing of the major divergence events. Because of the tradeoff between the amount of missing data and taxon/gene sampling, we reduced the initial matrix of 265 accessions and 12 loci to 95 accessions and 10 loci, and further to 42 species （and 7736 aligned nucleotides） to achieve stationary distributions in the Bayesian molecular clock runs. Results from a relaxed clock with an uncorrelated rates model and fossil-based calibration reveal that New World species are monophyletic and diverged from their mostly Asian sister clade some 30 mya, fitting with many other Beringian disjunctions. The split between the single North American and the single South American clade occurred approximately 25 mya, well before the closure of the Panamanian Isthmus. Overall, the biogeographic history of Ephedra appears dominated by long-distance dispersal, but finer-scale studies are needed to test this hypothesis.     

Molecular phylogeny of Soims-laubachia （Brassicaceae） s.l., based on multiple nuclear and plastid DNA sequences,and its biogeographic implications

The Hengduan Mountains region of south-west China is a noted biodiversity,hotspot, but the geographic origins and historical assembly of its rich endemic flora, including the sky-island species ofSolms-laubachia Muschl. （Brassicaceae）, have been little studied. Previous molecular studies on the phylogeny of Solms-laubachia showed it to be paraphyletic, leading to considerable expansion not only of its taxonomic limits, but also its geographic range, with the inclusion of taxa from outside the Hengduan region. However, these studies provided little resolution of interspecific relationships, preventing inferences about historical biogeography within the clade. In the present study, new sequence data from two nuclear genes （LEAFY and G3pdh） and two chloroplast intergenic spacers （petN-psbM and psbM-trnD） were combined with existing markers to increase phylogenetic signals. Phaeonychium villosum （Maxim.） Al-Shehbaz was found to be nested within Solms-laubachia s.l. In general, phylogenetic relationships appear to be a good predictor of geography, with the Hengduan Mountain endemics embedded in a paraphyletic grade of species from the western Himalayas and central Asia, but they also imply morphological homoplasy, lncongruence was detected between the nuclear and chloroplast gene trees, perhaps resulting from incomplete lineage sorting of ancestral polymorphisms. The crown age ofSolms-laubachia s.l. was estimated to be approximately 1.42-3.68 mya, using Bayesian relaxed molecular clock analysis. Historical biogeographic analysis using a parametric dispersalextinction-cladogenesis model inferred central Asia and the western Himalayas as most probable ancestral range of Solms-laubachia s.l., and estimated higher rates of eastward expansion than westward during the diversification of descendant lineages. In summary, our results suggest that Solms-laubachia s.l. originated during the Pliocene in central Asia, and subsequently migrated eastward into the Hengduan Mountains, colonizing sky-island, alpine scre     

Molecular phylogeny reveals the non-monophyly of tribe Yinshanieae(Brassicaceae) and description of a new tribe,Hillielleae

The taxonomic treatment within the unigeneric tribe Yinshanieae(Brassicaceae) is controversial, owing to differences in generic delimitation applied to its species. In this study, sequences from nuclear ITS and chloroplast trn L-F regions were used to test the monophyly of Yinshanieae, while two nuclear markers(ITS, ETS) and four chloroplast markers(trnL-F, trn H-psbA, rps16, rpL32-trnL) were used to elucidate the phylogenetic relationships within the tribe. Using maximum parsimony, maximum likelihood, and Bayesian inference methods, we reconstructed the phylogeny of Brassicaceae and Yinshanieae. The results show that Yinshanieae is not a monophyletic group, with the taxa splitting into two distantly related clades: one clade contains four taxa and falls in Lineage I, whereas the other includes all species previously placed in Hilliella and is embedded in the Expanded Lineage II. The tribe Yinshanieae is redefined, and a new tribe, Hillielleae, is proposed based on combined evidence from molecular phylogeny, morphology, and cytology.