A Molecular Phylogeny of Warbling-Finches (Poospiza): Paraphyly in a Neotropical Emberizid Genus

We investigated the phylogenetic relationships of 12 species within a single genus of neotropical passerine (Poospiza) using 849 bp (283 codons) of the cytochrome b mitochondrial gene. We further explored evolutionary affinities of these taxa using sequence from an additional 47 thraupine (tanagers) and 7 emberizine (sparrows and buntings) genera, members of the predominantly New World family Emberizidae. Poospiza have traditionally been considered part of the emberizine radiation. However, our analyses suggest that members of this genus are more closely related to some thraupine lineages than they are to the other neotropical emberizine genera included in our study (Atlapetes, Embernagra, Melopyrrha, Phrygilus, Saltatricula, Tiaris). Although member taxa are closely related, the genus Poospiza appears to be paraphyletic with representatives of 6 thraupine genera (Cnemoscopus, Cypsnagra, Hemispingus, Nephelornis, Pyrrhocoma, Thylpopsis) interspersed among four well-supported Poospiza clades. The majority of species within this Poospiza–thraupine clade have geographic ranges that are exclusive to, or partially overlap with, the Andes Mountains. It is probable that these mountains have played an important role in driving cladogenesis within this group. Sequence divergence (transversions only; mean 4.7 ± 1.3%) within the clade suggests that much of this diversification occurred within the late Miocene and Pliocene, a period coincident with major orogenic activity in central-western South America.     

Surveillance for batrachochytrium dendrobatidis using Mixophyes (Anura: Myobatrachidae) larvae

Fourteen populations of anuran larvae (tadpoles), including three populations of the endangered Fleay's Barred Frog (Mixophyes fleayi) and 11 populations of the common Great Barred Frog (Mixophyes fasciolatus), in creek sites in the southeast region of Queensland were selected. Site selection was based on a history (within the district) of adult frog population declines and/or disappearances or records of infection of adult frogs or larvae by Batrachochytrium dendrobatidis. Larvae were collected once from each creek site between October 2002 and October 2004, and were between Gosner developmental stages 25 and 40. Total body length ranged from 18 mm to 100 mm. Mouthparts were examined under a dissecting microscope for grossly visible abnormalities, and then examined for histologic evidence of B. dendrobatidis. The most consistent mouthpart abnormalities found were multifocal depigmentation of the jaw sheaths and loss or shortening of the tooth rows. At the individual larva level, presence of mouthpart abnormalities was strongly associated with histologic diagnosis of B. dendrobatidis (93%). At least one larva with abnormal mouthparts was detected at 12 of the 14 sites and histologic evidence of B. dendrobatidis was detected at 13 of the 14 sites. These findings suggest that larvae of Mixophyes species are suitable for surveillance for B. dendrobatidis. We conclude that surveillance of B. dendrobatidis where individual larva prevalences of mouthpart abnormalities and histologic evidence of B. dendrobatidis are as high as those observed in this study (66% and 78%, respectively), relatively small numbers of larvae are required to detect these infections. Medium to large larvae (body length >30 mm) were much more likely to be affected than small larvae (body length < or =30 mm), suggesting that larger individuals should be targeted for surveillance.     

Molecular Phylogeny of the Genus Paramesotriton (Caudata: Salamandridae)

To elucidate the phylogeny of the genus Paramesotriton (Caudata: Salamandridae), we investigated three mitochondrial DNA gene fragments (1207 bp in total) of cytochrome b, ND2, and ND4 for its six recognized species. The phylogenetic relationships within Paramesotriton were reconstructed by maximum parsimony (MP) and maximum likelihood (ML) methods. Phylogenetic trees (MP and ML trees) that were constructed from the combined data set of the three gene fragments indicated that all six species of Paramesotriton formed a monophyletic group, with P. caudopunctatus as basal to the other five species. This result suggests that P. fuzhongensis is a valid species in Paramesotriton.     

Soil pH influences embryonic survival in Pseudophryne bibronii (Anura: Myobatrachidae)

Abstract: There has been limited research examining the role that terrestrial habitat characteristics play in influencing persistence of amphibian populations. In this study we investigated the influence of a terrestrial habitat attribute on the distribution of the terrestrial egg‐laying toadlet, Pseudophryne bibronii. Eggs of this species are deposited in depressions, or under leaf litter, and develop to a stage where they can hatch to free swimming tadpoles when water covers breeding sites or embryos are washed into water. Because rainfall can be intermittent, eggs may sit for extended periods in terrestrial nests before sufficient rainfall initiates hatching. Appropriate egg‐laying sites must be chosen by both males and females to ensure embryos survive these periods. A study of 20 sites found that soil pH at sites where P. bibronii were recorded as present was significantly lower than pH at sites where P. bibronii were recorded as absent. To determine whether soil pH influenced the hatching success in P. bibronii, experiments were conducted in which batches of eggs were translocated to local areas with different soil pH. Survival rate of eggs was significantly influenced by relatively small changes in pH, with hatching success higher at lower pH. In a laboratory trial survival of eggs raised on sterilized soil was not affected by soil pH. Fungal infection of eggs was observed in the field and laboratory suggesting that hatching success might be affected via a complex interaction between soil pH, fungi and other soil biota. Choice trials using P. bibronii metamorphs indicated that individuals were capable of distinguishing between small differences in pH. Therefore, distribution of P. bibronii may be influenced by either differential mortality of embryos, or habitat choice by post metamorphic individuals, and further experiments are required to determine the relative importance of these factors.     

Synchronous polyandry and multiple paternity in the frog Crinia georgiana (Anura: Myobatrachidae)

Multiple paternity has rarely been reported in anuran amphibians, with only three previous documented examples. For the Australian frog Crinia georgiana, we observed synchronous polyandry in an average of 44% of matings observed at four field sites. This suggests matings involving more than one male are common in this species. One to eight males were observed in amplectant groups with second males amplexed ventrally. Genetic analyses, using allozyme electrophoresis, of offspring from two matings indicated that at least two of three possible males fathered offspring. Third males were unlikely to have shared paternity, explained by their inappropriate position during amplexus. Multiple paternity may be more common in frogs than has been reported. Copyright 1999 The Association for the Study of Animal Behaviour.     

Molecular Phylogeny and Biogeography of the Genus Ilex L. (Aquifoliaceae)

The chloroplast atpB-rbcL spacer was sequenced for 115 speciesof Ilex and forNemopanthus mucronatus (Aquifoliaceae). In addition,rbcL sequences were obtained for 47 selected species. Takentogether with fossil records in the literature, the data indicatethat: (1) the lineage Ilex was already cosmopolitan long beforethe end of the Cretaceous. A relative test of the rate of nucleotidesubstitution indicates that extant species do not representthe entire lineage because of the extinction of its basal branches.The common ancestor of the extant species probably appearedat the upper Tertiary. (2) Several Asian/North American disjunctionsare observed, as well as North American/South American relationships.The directions of these relationships are yet to be determined.The African species Ilex mitis is closely related to Asian species.Ilex anomala (Hawaii and Tahiti) is related to American species.Ilex perado and Ilex canariensis (both in Macaronesia) havetotally different relationships, the former being related toEurasian species, while relationships of the latter remain obscure.(3) Gene trees substantially contradict the systematics of Loesener.Nemopanthus,closely related to Ilex amelanchier, is nested in the genusIlex. The family Aquifoliaceae is organized in four groups,each of them having a geographic or ecological peculiarity.Copyright 2000 Annals of Botany Company Aquifoliaceae, chloroplast DNA, fossil record, historical biogeography, Ilex (holly tree), molecular phylogeny.     

Comparative skeletogenesis of the Oriental Tree Frog Hyla orientalis (Anura: Hylidae)

The ontogeny of skeletal development of the Oriental Tree Frog, Hyla orientalis (Bedriaga, 1890), is followed from tadpole to froglet and compared with four other hylid species i.e., Hypsiboas lanciformis, Hypsiboas pulchellus, Phyllomedusa vaillanti and Pseudis platensis. Our analysis and comparison is based on cleared and double-stained specimens. The parasphenoid is the first skeletal element that ossified in Hyla orientalis is followed by the exoocipital, prootic, axial and appendicular skeletal elements, frontoparietal, nasal, upper jaw, squamosal, pterygoid, lower jaw, vomer and quadratojugal. Ossification initiated at early Stage 33 in Hyla orientalis whereas in other hylids it begins after Stage 37. Major transformations of the larval jaw were visible after Stage 42. Cranial ossifications were completed after appendicular and axial ossifications of Hyla orientalis. All skeletal elements ossified before the end of the metamorphosis, a unique feature of Hyla orientalis in comparison to the other hylids studied here.     

Molecular phylogeny of the Australian frog genera Crinia, Geocrinia, and allied taxa (Anura: Myobatrachidae).

We present a mitochondrial gene tree for representative species of all the genera in the subfamily Myobatrachinae, with special emphasis on Crinia and Geocrinia. This group has been the subject of a number of long-standing taxonomic and phylogenetic debates. Our phylogeny is based on data from approximately 780 bp of 12S rRNA and 676 bp of ND2, and resolves a number of these problems. We confirm that the morphologically highly derived monotypic genera Metacrinia, Myobatrachus, and Arenophryne are closely related, and that Pseudophryne forms the sister group to these genera. Uperoleia and the recently described genus Spicospina are also part of this clade. Our data show that Assa and Geocrinia are reciprocally monophyletic and together they form a well-supported clade. Geocrinia is monophyletic and the phylogenetic relationships with the genus are fully resolved with two major species groups identified: G. leai, G. victoriana, and G. laevis; and G. rosea, G. alba, and G. vitellina (we were unable to sample G. lutea). We confirm that Taudactylus forms the sister group to the other myobatrachine genera, but our data are equivocal on the phylogenetic position of Paracrinia. The phylogenetic relationships among Crinia species are well resolved with strong support for a number of distinct monophyletic clades, but more data are required to resolve relationships among these major Crinia clades. Crinia tasmaniensis and Bryobatrachus nimbus form the sister clade to the rest of Crinia. Due to the lack of generic level synapomorphies for a Bryobatrachus that includes C. tasmaniensis, we synonymize Bryobatrachus with Crinia. Crinia georgiana does not form a clade distinct from other Crinia species and so our data do not support recognition of the genus Ranidella for other Crinia species. Crinia subinsignifera, C. pseudinsignifera, and C. insignifera are extremely closely related despite differences in male advertisement call. A preliminary investigation of phylogeographic substructure within C. signifera revealed significant divergence between samples from across the range of this species.     

A New Species of the Genus Rhacophorus(Anura:Rhacophoridae) from Southern China

A new species of rhacophorid of the genus Rhacophorus is described from the Shiwandashan National Nature Reserve, Guangxi, Southern China. Rhacophorus pinglongensis sp. nov. is compared with congeners from China and other parts of Southeast Asia. The new species is distinguished from its congeners by combination of following characters:(1) small size(adult male, SVL 32.0-38.5 mm);(2) smooth and green dorsum;(3) flanks, axilla, ventral surface of forearms, inguinal, anterior and posterior surfaces of thighs, ventral surface of shank, and dorsal surface of feet covered with black blotches with white spots or white spots with a faint orange tint;(4) ventral surface of feet and webbing tangerine;(5) supratympanic fold weak;(6) outer margin of forearms and feet with low dermal ridges;(7) calcars absent on heels;(8) iris silver, diffusing to ecru laterally with light khaki ring along margin. The new species is closely related to R. dorsoviridis, R. moltrechti, and R. nigropunctatus based on adult morphology. Currently, this species is known only from mid-elevation montane evergreen forest in the Shiwandashan National Nature Reserve, Guangxi, China.     

A New Species of the Genus Leptolalax(Anura: Megophryidae) from Northern Thailand

A new species of the genus Leptolalax is described from Doi Saket, Chiang Mai Province, northern Thailand. The new species is distinguished from its congeners by the following combination of characters: 1) adult size large(male SVL 45.8-52.5 mm); 2) tympanum distinct; 3) skin of dorsal surface mostly smooth, with distinct tubercles on dorsal and lateral parts of the body; 4) ventrolateral glandular ridge distinct and complete; 5) lateral body and groin covered with dark brown or black spots; 6) throat, chest, and belly nearly immaculate white; and 7) bi-colored iris, upper onethird golden, and lower two-thirds gray. Molecular phylogenetic analysis of mitochondrial 16S rRNA further distinguishes it from its congeners for which sequences are available.     

A Molecular Phylogeny of the Aphidiinae (Hymenoptera: Braconidae)

Phylogenetic relationships within the Aphidiinae, and between this and other subfamilies of Braconidae (Hymenoptera), were investigated using sequence data from three genes: elongation factor-1α, cytochrome b, and the second expansion segment of the 28S ribosomal subunit. Variation in both protein-coding genes was characterized by a high level of homoplasy, but analysis of the expansion segment—robust over a range of alignment methods and parameters—resolved some of the older divergences. Parsimony analysis of the combined data suggests the following tribal relationships: (Ephedrini + (Praini + (Aphidiini + Trioxini))). In addition, the cyclostome subfamilies were found to form a clade separate from the Aphidiinae, but relationships between the Aphidiinae and the noncyclostome braconids could not be resolved. The inferred phylogeny also supported a secondary loss of internal pupation within the Praini and a polyphyletic origin of endoparasitism within the Braconidae.     

Phylogeny of West AfricanCaryedon(Coleoptera: Bruchidae): Congruence between Molecular and Morphological Data

Seed beetles belonging to the Old World genusCaryedonfeed in the seeds of various Caesalpinioideae, Mimosoideae, and Combretaceae. In an attempt to resolve broad phylogenetic relationships within the genus, we obtained 332 base pair sequences of mitochondrial 12S ribosomal DNA and morphological data for the 16 West AfricanCaryedonspecies. Morphological characters were analyzed under maximum parsimony and sequences were compared under maximum parsimony, maximum likelihood, and neighbor joining. Using a partition homogeneity test, we determined that morphological and molecular data sets were combinable. Combined data were analyzed under maximum parsimony. Morphological and molecular trees were congruent at the species group level and total evidence analyses yielded the same topologies as molecular data with each of the three outgroups used. Four main terminal clades are recognized, each corresponding with a group of species generally feeding on the same host plant family, subfamily, genus, or species. The monophyly of legume feedingCaryedonis supported by both data sets, and Combretaceae feeders split in two monophyletic assemblages.     

Molecular Phylogeny and Historical Biogeography of the Neotropical Swarm-Founding Social Wasp Genus Synoeca (Hymenoptera: Vespidae)

The Neotropical Region harbors high biodiversity and many studies on mammals, reptiles, amphibians and avifauna have investigated the causes for this pattern. However, there is a paucity of such studies that focus on Neotropical insect groups. Synoeca de Saussure, 1852 is a Neotropical swarm-founding social wasp genus with five described species that is broadly and conspicuously distributed throughout the Neotropics. Here, we infer the phylogenetic relationships, diversification times, and historical biogeography of Synoeca species. We also investigate samples of the disjoint populations of S. septentrionalis that occur in both northwestern parts of South America through Central American and the Brazilian Atlantic rainforests. Our results showed that the interspecific relationships for Synoeca could be described as follows: (S. chalibea + S. virginea) + (S. cyanea + (S. septentrionalis/S. surinama)). Notably, samples of S. septentrionalis and S. surinama collected in the Atlantic Forest were interrelated and may be the result of incomplete lineage sorting and/or mitochondrial introgression among them. Our Bayesian divergence dating analysis revealed recent Plio-Pleistocene diversification in Synoeca. Moreover, our biogeographical analysis suggested an Amazonian origin of Synoeca, with three main dispersal events subsequently occurring during the Plio-Pleistocene.     

中国四川省蛙科一新种——南江臭蛙(两栖纲:无尾目)

对分布于四川、重庆市、贵州、湖北、湖南、安徽和福建的花臭蛙[Odorrana(Odorrana)schmackeri(Boettger,1892)]标本进行了形态比较,结果发现四川省南江和万源县的花臭蛙标本与其他产地的花臭蛙标本在形态特征上存在明显区别,主要是:1)成体雄雌体长之比值较小,约为1∶1.34;2)犁骨齿列短,两内侧间距宽,距内鼻孔远;3)背面深色斑点周围无浅色边缘;4)股后部深色斑大而稀疏;5)趾间蹼较弱,第四趾蹼达远端关节下瘤;6)腹内成熟卵的动植物极均为乳白色。故将四川省南江和万源的标本订为新种--南江臭蛙[Odorrana(Odorrana)nanjiangensis,sp.nov.]。     

东秦岭地区齿突蟾属一新种记述（两栖纲：角蟾科）

2020年5月在洛阳市嵩县白云山国家森林公园玉皇顶林场（33°38′37″N,111°50′01″E,海拔1 882 m）采集到齿突蟾属（Scutiger）物种9号标本,经形态特征比较及线粒体COI和Cyt b基因联合构建的齿突蟾属部分物种系统发育树显示,采集到的齿突蟾标本系一新种,命名为华夏齿突蟾Scutiger feiliangi Zhou, Guan and Shi, sp. nov.。其主要鉴别特征为：1）体型中等,雄性体长45.7～50.2 mm,雌性体长48.9～51.5 mm;2）头宽略大于头长,鼓膜不显;3）前臂及手长略超过体长之半,后肢较短;4）背面粗糙,布满疣粒;5）指间无蹼,趾间微蹼;6）具两对胸腺,外侧者略小于内侧;7）背部有4行断续排列的纵行肤褶;8）肛周有多个近圆形腺体。同时,对新种华夏齿突蟾的生物学特性进行描述。     

Molecular Phylogeny and Species Identification of Pufferfish of the Genus Takifugu (Tetraodontiformes, Tetraodontidae)

The phylogenetic relationships and species identification of pufferfishes of the genus Takifugu were examined by use of randomly amplified polymorphic DNA (RAPD) and sequencing of the amplified partial mitochondrial 16S ribosomal RNA genes. Amplifications with 200 ten-base primers under predetermined optimal reaction conditions yielded 1962 reproducible amplified fragments ranging from 200 to 3000 bp. Genetic distances between 5 species of Takifugu and Lagocephalus spadiceus as the outgroup were calculated from the presence or absence of the amplified fragments. Approximately 572 bp of the 16S ribosomal RNA gene was amplified, using universal primers, and used to determine the genetic distance values. Topological phylogenic trees for the 5 species of Takifugu and outgroup were generated from neighbor-joining analysis based on the data set of RAPD analysis and sequences of mitochondrial 16S rDNA. The genetic distance between Takifugu rubripes and Takifugu pseudommus was almost the same as that between individuals within each species, but much smaller than that between T. rubripes, T. pseudommus, and the other species. The molecular data gathered from both analysis of mitochondria and nuclear DNA strongly indicated that T. rubripes and T. pseudommus should be regarded as the same species. A fragment of approximately 900 bp was amplified from the genome of all 26 T. pseudommus individuals examined and 4 individuals of intermediate varieties between T. rubripes and T. pseudommus. Of the 32 T. rubripes individuals, only 3 had the amplified fragment. These results suggest that this fragment may be useful in distinguishing between T. rubripes and T. pseudommus. Received September 29, 2000; accepted February 26, 2001.