Comparative morphology of pond,stream and phytotelm‐dwelling tadpoles of the South American Redbelly Toads (Anura: Bufonidae: Melanophryniscus)

We present a comprehensive review of larval morphology in the Neotropical toad genus Melanophryniscus. The taxa studied included 23 species with representatives of recognized phenetic groups and different larval ecomorphological guilds: pond, stream, and phytotelm‐dwelling tadpoles. Their external morphology variation is congruent with current phenetic arrangement based on adult features, but also reflects the habitat where larvae develop. Lotic tadpoles (i.e. M. tumifrons group and M. krauczuki) in general exhibit a more depressed body, a longer tail with lower fins, and larger oral discs than lentic forms (i.e. M. stelzneri group, M. moreirae, M. sanmartini, and M. langonei). Despite their peculiar, confined microhabitat, phytotelm larvae do not diverge markedly from non‐arboreal species. The distinctive features of all species are the presence of a pineal end organ and the placement of the intestinal reversal point at the left of the abdomen in typical larval stages. The buccal cavity and musculoskeletal anatomy are quite conserved between species, yet some characteristics differ from those of other bufonids. The presence of one pair of subhyoid muscles is apparently an exclusive trait of Melanophryniscus among Bufonidae. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 112 , 417–441.     

Ontogeny, phylogeny, and morphology in anuran larvae: morphometric analysis of cranial development and evolution in Rana tadpoles (Anura: Ranidae)

Comparative studies of chondrocranial morphology in larval anurans are typically qualitative in nature, focusing primarily on discrete variation or gross differences in the size or shape of individual structures. Detailed data on chondrocranial allometry are currently limited to only two species, Rana sylvatica and Bufo americanus. This study uses geometric morphometric and multivariate statistical analyses to examine interspecific variation in both larval chondrocranial shape and patterns of ontogenetic allometry among six species of Rana. Variation is interpreted within the context of hypothesized phylogenetic relationships among these species. Canonical variates analyses of geometric morphometric datasets indicate that species can be clearly discriminated based on chondrocranial shape, even when whole ontogenies are included in the analysis. Ordinations and cluster analyses based on chondrocranial shape data indicate the presence of three primary groupings (R. sylvatica; R. catesbeiana + R. clamitans; and R. palustris + R. pipiens + R. sphenocephala), and patterns of similarity closely reflect phylogenetic relationships. Analysis of chondrocranial allometry reveals that some patterns are conserved across all species (e.g., most measurements scale with negative allometry, those associated with the posterior palatoquadrate tend to scale with isometry or positive allometry). Ontogenetic scaling along similar allometric trajectories, lateral transpositions of individual trajectories, and variable allometric relationships all contribute to shape differences among species. Overall patterns of similarity among ontogenetic trajectories also strongly reflect phylogenetic relationships. Thus, this study demonstrates a tight link between ontogeny, phylogeny, and morphology, and highlights the importance of including both ontogenetic and phylogenetic data in studies of chondrocranial evolution in larval anurans.     

Living in fast‐flowing water: Morphology of the gastromyzophorous tadpole of the bufonid Rhinella quechua (R. veraguensis group)

We describe the bufonid gastromyzophorous tadpoles of Rhinella quechua from montane forest streams in Bolivia. Specimens were cleared and stained, and the external morphology, buccopharyngeal structures, and the musculoskeletal system were studied. These tadpoles show a combination of some traits common in Rhinella larvae (e.g., emarginate oral disc with large ventral gap in the marginal papillae, labial tooth row formula 2/3, prenarial ridge, two infralabial papillae, quadratoorbital commissure present, larval otic process absent, mm. mandibulolabialis superior, interhyoideus posterior, and diaphragmatopraecordialis absent, m. subarcualis rectus I composed of three slips), some traits apparently exclusive for the described species of the R. veraguensis group (e.g., second anterior labial tooth row complete, lingual papillae absent, adrostral cartilages present), and some traits that are shared with other gastromyzophorous tadpoles (e.g., enlarged oral disc, short and wide articular process of the palatoquadrate, several muscles inserting on the abdominal sucker). In the context of the substantial taxonomic and nomenclatural changes that the former genus Bufo has undergone, and despite the conspicuous morphological differences related to the presence of an abdominal sucker, the larval morphology of R. quechua supports including it in the genus Rhinella and placing it close to species of the R. veraguensis assemblage. J. Morphol., 2009. © 2009 Wiley‐Liss, Inc.     

Morphology of the cranial skeleton and musculature in the obligate carnivorous tadpole of Lepidobatrachus laevis (Anura: Ceratophryidae)

Ziermann, J.M., Infante, C., Hanken, J. and Olsson, L. 2011. Morphology of the cranial skeleton and musculature in the obligate carnivorous tadpole of Lepidobatrachus laevis (Anura: Ceratophryidae). —Acta Zoologica (Stockholm) 00 :1–12. Lepidobatrachus laevis (Ceratophryidae: Ceratophryinae) is a bizarre frog endemic to the Chacoan desert of central South America. Its tadpole is an obligate carnivore that can catch and consume live prey nearly its own size. Morphological adaptations associated with this unique feeding mode, including the larval skull anatomy and associated cranial musculature, have only been partly described. We studied the head of Stages 26–27 larvae using gross dissection, immunohistochemistry, and standard histology. Derived features of this tadpole compared to the microphagous, herbivorous larvae of most other anurans include simplified chondrocranial cartilages and very robust jaw muscles. The mm. suspensorio‐ et quadratoangularis do not take their origin from the processus muscularis of the palatoquadrate, as in most other tadpoles, but instead originate from the corpus of the palatoquadrate caudal to this process. The jaw levators are unusually large. The tadpole of Ceratophrys, another member of the ceratophryine clade, also consumes large animal prey, but its morphology is very different. It probably has evolved independently from a generalized, mainly herbivorous tadpole similar to the larva of Chacophrys, the third ceratophryine genus. Most specialized features of the larval head of Lepidobatrachus laevis are adaptations for ‘megalophagy’—ingestion of whole, very large animal prey.     

Chondrocranial morphology of Leptodactylus larvae (Leptodactylidae: Leptodactylinae): Its utility in phylogenetic reconstruction

Chondrocranial morphology of leptodactylid frogs is scarcely known and has not been completely described for any species of Leptodactylus. We describe the diversity of chondrocranial morphology in the genus Leptodactylus based on the analysis of 22 species, representing the four species groups: the fuscus Group, ocellatus Group, melanonotus Group, and pentadactylus Group. Furthermore, 26 characters are identified and used in a phylogenetic analysis. The phylogenetic analysis using Physalaemus, Crossodactylus, and Hylodes as outgroups suggests two monophyletic clades within Leptodactylus: the melanonotus-ocellatus clade and the pentadactylus-fuscus clade. However, it does not support the monophyly of the species groups as currently recognized and it suggests a paraphyletic Leptodactylus. Enforcing the monophyly of the ingroup, i.e., Leptodactylus, results in the same major two clades of Leptodactylus. Leptodactylus riveroi, a taxon previously unassigned to any species group, appears most closely related to the melanonotus-ocellatus clade based on chondrocranial characteristics. J. Morphol. 238:287–305, 1998. © 1998 Wiley-Liss, Inc.     

Positive Darwinian selection results in resistance to cardioactive toxins in true toads (Anura: Bufonidae)

Members of the Family Bufonidae, true toads, are famous for their endogenously synthesized cardioactive steroids that serve as defensive toxins. Evolution of resistance to these toxins is not understood. We sequenced a key region of the toxin''s binding site in the Na⁺/K⁺ ATPase for relevant taxa representing Hyloidea (including bufonids), Ranoidea and Archaeobatrachia and tested for positive selection in a phylogenetic context. Bufonidae were distinct from other Hyloidea at 4–6 of 12 sites and, with one exception, had a homologous amino acid sequence. Melanophryniscus stelzneri had a distinct sequence, consistent with other independent evidence for a differentiated toxin. Tests within Bufonidae detected positive selection within the binding region, providing, to our knowledge, the first evidence of this type for positive selection within Amphibia. There was no evidence for positive selection on Bufonidae or M. stelzneri lineages. Sequence change in Leptodactylus ocellatus, a leptodactylid predator of Bufonidae, provides a molecular basis for predator resistance possibly associated with gene duplication.     

A new sinistral species of the land-snail genus Satsuma (Pulmonata: Camaenidae) from China

ABSTRACT

The species of Satsuma are mostly endemic to East Asia except for one species distributed in Batan Island of the Philippines. More than 99% of the known species of this genus are endemic to the island environment. Only three species are currently known to occur on the mainland. Here we describe a new sinistral Satsuma species from China, Satsuma guandi n. sp. We studied the shell morphology and genital anatomy of the new species and reconstructed the molecular phylogeny of the genus based on partial nucleotide sequences of the mitochondrial gene cytochrome c oxidase subunit I and nuclear markers from the ribosomal RNA cistrons (the internal transcribed spacer and the external transcribed spacer regions). The new species differs from other sinistral Satsuma species by having a strongly angulated shell and an open dark brownish-red umbilicus. The new species also differed from all other sinistral congeners in details of its reproductive anatomy. The molecular analysis supports the validity of the new species within the genus Satsuma.     

Isolation and high genetic diversity in dwarf mountain toads (Capensibufo) from South Africa

Traditional models of amphibian dispersal and gene flow point to low dispersal and high philopatry. In recent years, this traditional view has been challenged and it appears that no general model holds across taxa. Conservation of amphibians cannot be addressed on an over‐arching scale, but must come on a case‐by‐case basis, especially for range‐restricted species where information on gene flow and migration must be incorporated into conservation efforts. The only two members of the genus Capensibufo Grandison, 1980 (Anura: Bufonidae) are range restricted small bufonids, with distributions limited to montane areas in South Africa. Using a Bayesian analysis of two mitochondrial markers (16S and ND2), we examined the genetic patterns in Capensibufo rosei and Capensibufo tradouwi in order to understand both taxonomic and geographic boundaries. These species were not monophyletic, and demonstrate no clear taxonomic boundaries. Instead, the genus is extremely diverse genetically, with distinct lineages confined to isolated mountains that represent geographic boundaries. In addition, bioclimatic modelling using MAXENT and scenarios of climatic conditions at both the present and last glacial maximum suggest multiple bioclimatic and physical barriers to gene flow at present and in the past. We conclude that members of the genus have very low vagility, that current taxonomic boundaries are inadequate, and that strong geographic structuring has undoubtedly contributed to genetic diversity at the species level, rather than the population level. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 822–834.     

Patterns of fluctuating asymmetry in the limbs of anurans

It has been hypothesized that fluctuating asymmetry (FA) may provide an indication of the functional importance of structures within an organism, with structures that more strongly impact fitness being more symmetric. Based on this idea, we predicted that for tetrapods in which the forelimbs and hindlimbs play an unequal role in locomotion, the less functionally important limb set should display higher levels of FA. We conducted a multispecies test of this hypothesis in anurans (frogs and toads), whose saltatory locomotor mode is powered by the hindlimbs. We also tested whether FA in the forelimbs, which play a more important role during landing, differed between families that differ in the degree of forelimb use in locomotion (Bufonidae vs. Ranidae). We calculated FA from the lengths of humeri and femora measured from disarticulated skeletal specimens of four anuran taxa (Bufonidae: Anaxyrus americanus, Rhinella marina; Ranidae: Lithobates catesbeianus, Lithobates clamitans). Our findings were consistent with the hypothesis that natural selection for increased locomotor performance may influence patterns of FA seen in vertebrate limbs, with all species displaying lower mean FA in the hindlimbs. More subtle functional roles between the forelimbs of bufonids and ranids, however, did not elicit different levels of FA.     

Larval stages of the crab Mithrax tortugae (Brachyura: Mithracidae) with comparisons between all species of Mithrax

The postembryonic larval stages of Mithrax tortugae are described, illustrated and compared with the described zoeae of other Mithrax species. Mithrax tortugae showed morphological features in all the stages of larval development that differed from those observed in other species of Mithrax, especially M. hispidus. In the Zoea I stage, M. tortugae and M. pleuracanthus lacked the minute spine on the dorsal spine observed in M. hispidus; M. tortugae exhibited a terminal spine on the inner lobe of the coxal endite of the maxilla, which was not observed in M. hispidus or M. pleuracanthus. Also, M. tortugae exhibited furcae with spines that are not spinulated, whereas in M. hispidus and M. pleuracanthus these spines are spinulated. In the Zoea II stage, M. tortugae showed a terminal spine on the coxal endite of the maxillula, whereas in M. hispidus and M. pleuracanthus this spine is absent. In the Megalopa stage, we also observed differences in the sternal plate setation between M. tortugae and M. hispidus, where M. tortugae had eight simple setae and M. hispidus showed two simple and four plumodenticulate setae. Partial sequences of the 16S rRNA and COI genes of the parental female were analysed, providing additional evidence for species identification. Together, our analyses of larval morphology and the results of the molecular analyses reinforced recognition of the relationships among M. tortugae, M. hispidus and M. pleuracanthus.     

Phylogeny of the Malus (Apple Tree) Species,Inferred from the Morphological Traits and Molecular DNA Analysis

Some debated issues of the genus Malus (apple) taxonomy were examined using a variety of species from the collection of the Maikop Experimental Station, Vavilon Research Institute of Plant Industry (Krasnodar krai). Phylogenetic relationships among these species were studied using traditional analysis of morphological traits, RAPD, and complete sequencing of the 5"- internal transcribed spacer (ITS1), 5.8S rRNA, 3"- internal transcribed spacer (ITS2) (constituting a cluster of the rRNA genes), and the terminal fragment of the matK gene encoding chloroplast maturase. The results showed that the Sorbomalussection was polyphyletic; the American apple M. fusca was closely related to the species contributing to the East Asian center of the genus origin, and the American speciesM. angustifolia, M. coronaria, and M. ioensis were closely related to the M. trilobata relict species, whose assignment to the genus Malus is debated by some authors. Molecular analysis of the species relationships showed that the Middle Asian apple M. sieversii is the species from which apple domestication started.     

Around the world in 10 million years: biogeography of the nearly cosmopolitan true toads (Anura: Bufonidae)

Aim The species‐rich family of true toads (Anura: Bufonidae) has been the focus of several earlier studies investigating the biogeography of geographically widespread taxa. Herein, we employ newly developed Bayesian divergence estimate methods to investigate the biogeographical history of this group. Resulting age estimates are used to test several key temporal hypotheses including that the origin of the bufonid clade pre‐dates Gondwanan vicariance (~105 million years ago, Ma). Area cladograms are also invoked to investigate the geographical origin of the family. Location Worldwide, except the Australia–New Guinea plate, Madagascar and the Antarctic. Methods A phylogenetic hypothesis of the relationships among true toads was derived from analysis of 2521 bp of DNA data including fragments from three mitochondrial (12S, tRNA^val, 16S) and two nuclear (RAG‐1, CXCR‐4) genes. Analysis of multiple, unlinked loci with a Bayesian method for estimating divergence times allowed us to address the timing and biogeographical history of Bufonidae. Resulting divergence estimates permitted the investigation of alternative vicariance/dispersal scenarios that have been proposed for true toads. Results Our area cladogram resulting from phylogenetic analysis of DNA data supports a South American origin for Bufonidae. Divergence estimates indicate that the family originated earlier than had been suggested previously (78–99 Ma). The age of the enigmatic Caribbean clade was dated to the late Palaeocene–early Eocene. A return of bufonids to the New World in the Eocene was followed by rapid diversification and secondary expansion into South America by the early Oligocene (Rupelian). Main conclusions The South American origin of Bufonidae in the Upper Cretaceous was followed by relatively rapid expansion and radiation around the globe, ending with a return to the Americas via a Eurasian/North American land bridge in the Eocene. Though the exact route of this dispersal (Beringia or North Atlantic) remains unclear, an argument is made for the less frequently invoked North Atlantic connection. The origin of the enigmatic Caribbean lineage was found to be consistent with colonization following the bolide impact at the K/T boundary. These findings provide the first, firm foundation for understanding true toad divergence times and their truly remarkable and global radiation.     

A taxonomic study of Whitehead's torrent frog,Meristogenys whiteheadi,with descriptions of two new species (Amphibia: Ranidae)

The genus Meristogenys (Anura: Ranidae), endemic to Borneo, presents serious taxonomic problems despite being one of the commonest frogs in the mountainous regions of this island. We investigated molecular and morphological variations in Meristogenys whiteheadi (Boulenger, 1887) using larval and adult specimens from Sabah and Sarawak (Malaysia). We found three allopatric lineages in this species. We regard each of these as a distinct species because they are separated by a large genetic distance, and do not form any monophyletic group. Their morphological characters indicate that the distributional range of M. whiteheadi s.s. is divided into two disjunct areas: Mt Kinabalu (northern Sabah) and northern Sarawak. The two other lineages occupy ranges between those of M. whiteheadi, and represent undescribed cryptic species. One of these, Meristogenys stigmachilus sp. nov. , collected from the northern part of the Crocker Range, is distinguished from M. whiteheadi by black spots on the upper lip and dark dots scattered on the back. A second undescribed species, Meristogenys stenocephalus sp. nov. , was collected mainly from the southern part of the Crocker Range, and is characterized by the large body size of males and a relatively narrow head. Meristogenys stenocephalus sp. nov. also differs from M. stigmachilus sp. nov. and M. whiteheadi in larval morphology, but larvae of the latter two cannot be differentiated morphologically. We discuss relative tibia length, a diagnostic specific characteristic in the genus Meristogenys, and the relationships between body size and sexual size dimorphism in this genus. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 161 , 157–183.     

Veratrum album and V. nigrum (Melanthiaceae) in Italy: Micromorphology and systematics

ABSTRACT

Floral anatomy and pollen morphology of the two European species of Veratrum (V. nigrum and V. album subsp. lobelianum) from Italy are described in the context of the systematics of the genus and tribe Melanthieae (Melanthiaceae sensu stricto). Septal nectaries are absent, as in other Liliales. Most characters in Veratrum are typical of Melanthieae: e.g. semi-inferior ovaries, fused carpellary bundles and operculate pollen.     

Phylogenetic relationships of bufonid frogs and tests of alternate macroevolutionary hypotheses characterizing their radiation

The frog family Bufonidae is a large group distributed throughout all major land masses of die world except the polar regions. Although the monophyly of Bufonidae is strongly supported, phylogenetic relationships within the group are not well understood. Because of apparently large differences in rates of morphological and behavioral evolution, speciation, and geographical range, Bufonidae raises intriguing macroevolutionary questions. For example, one might imagine that the group's evolutionary history is congruent (1) with its morphological and behavioral character distribution, or (2) with its current geographical positioning. To examine phylogenetic relationships within Bufonidae and test these alternate hypotheses, DNA sequence data were obtained from four genes, three mitochondrial and one nuclear, from populations throughout the geographic and taxonomic breadth of the family. Phylogenetic analyses of these sequences by several methods support several hypotheses of relationships, notably: (1) contrary to the previous hypotheses of some workers, Bufo is not the basal, wildly paraphyletic genus of Bufonidae, although it is also not monophyletic, (2) the basal splits within bufonids separate South American 'atelopodid'genera from all other bufonids, although the monophyly of the 'atelopodids' is not supported, and (3) the most highly supported clades within the family generally correspond to small- to medium-sized, geographically and morphologically homogeneous groups. These trees are used to test the two alternate macroevolutionary hypotheses, and, while neither hypothesis fully explains the observed phylogenetic relationships, these relationships are broadly consistent with both character distribution and geography at particular levels.