Parsimony Jackknifing Research Articles

Island life - classification, speciation and cryptic species ofPycnandra(Sapotaceae) in New Caledonia

Pycnandra (Sapotaceae), the largest endemic genus in New Caledonia, comprises 66 species classified in six subgenera. We tested phylogenetic relationships and a proposed infrageneric classification by sampling 60 species for sequences of nuclear ribosomal DNA (ETS, ITS, RPB2) and plastid DNA (trnH–psbA) and nine morphological characters. Data were analysed with Bayesian inference, parsimony jackknifing and lineage through time. We recovered a phylogenetic tree supporting the recognition of six proposed subgenera (Achradotypus, Leptostylis, Pycnandra, Sebertia, Trouettia and Wagapensia). Because a subgeneric classification is used, the nomenclature will be stable when the members are transferred to Pycnandra. Morphological traits were optimized in the BEAST analysis, adding evidence to earlier work that morphology has limited value for successfully diagnosing groups in Sapotaceae. We confirm a previously suspected case of cryptic species that exhibit the same morphological features and require the same abiotic conditions, but are distantly related in the phylogenetic tree. We detected two possible new cases of cryptic sibling species that might warrant recognition. A slowdown in speciation rate in several genera has been suggested as evidence that New Caledonia was once submerged after rifting from Australia. Plotting lineages through time reveals two important intervals at 7.5–8.6 Ma and present to 1.5 Ma, when net molecular diversification within the genus was zero. This indicates that the genus presently has reached a dynamic equilibrium, providing additional evidence that New Caledonia is an old Darwinian island, being submerged during the Eocene and colonized after re-emergence c. 37 Ma.

A new species, genus and tribe of Sapotaceae, endemic to Madagascar

AbstractPhylogenetic relationships of the two Malagasy Sapotaceae endemic genera Capurodendron and Tsebona have been unclear until now. Recent collections from Madagascar, as well as a better representation of the tribe Isonandreae, altogether 95 terminals, were used to estimate a phylogeny of subfamily Sapotoideae. We analysed sequences of nrDNA (ITS) and cpDNA (trnH‐psbA) with Bayesian inference and parsimony jackknifing. As in previous analyses, Sapoteae and Sideroxyleae are recovered monophyletic. In addition, Isonandreae, distributed in the Indo‐Pacific, is for the first time resolved as monophyletic and sister to Sapoteae. All Malagasy accessions of Capurodendron, Tsebona, and a new species are grouped in another well‐supported clade. This clade is accommodated in a new tribe Tseboneae characterized by caducous stipules, 5‐merous flowers with quincuncial sepals, contorted aestivation of corolla lobes, absence of corolla appendages, one or three stamens opposite each corolla lobe, villous staminodes, seeds with an adaxial scar and plano‐convex cotyledons, lacking endosperm. The new species is described in the new genus Bemangidia (B. lowryi) because it has a unique leaf venation for the tribe and combines different morphological features from Capurodendron and Tsebona. All three genera are well‐supported monophyletic groups. Bemangidia lowryi is threatened with extinction due to extensive ongoing forest destruction and is assigned a preliminary conservation status of Critically Endangered.

Towards a natural classification of Sapotaceae subfamily Chrysophylloideae in Oceania and Southeast Asia based on nuclear sequence data

AbstractGeneric limits within subfamily Chrysophylloideae (Sapotaceae) from Oceania and Southeast Asia are reconciled based on a molecular phylogeny. We analysed sequences of nuclear ribosomal DNA (ETS, ITS) and the nuclear gene RPB2 with BEAST and parsimony jackknifing, using a sample of 168 terminals. Eight morphological characters were traced on a condensed majority–rule consensus tree to identify diagnostic character combinations for the genera. Accepted genera with character support are Magodendron, Pichonia, Planchonella, Pycnandra, Sersalisia, and Van–royena, while Beccariella and Niemeyera require amendment. Beccariella, a widely distributed group, is an illegitimate later homonym and we propose that the genus Pleioluma is resurrected in its place. The Australian genus Niemeyera is paraphyletic, but it is rendered monophyletic by reinstating Amorphospermum for N. antiloga. Beauvisagea, Blabeia, Fontbrunea, and Krausella are all segregates of Planchonella and rejected, while Wokoia is a later synonym of Pichonia. Planchonella baillonii, an endemic species of New Caledonia, is the sole member of an old lineage and firmly placed as the sister to a clade comprising the other congeners. Planchonella sandwicensis, a Hawaiian species, previously proposed to be a distinct genus, is a member of Planchonella. In the Pacific, P. tahitensis (including P. grayana) is a polymorphic species, widely distributed and adapted to a wide range of habitats. We provide a generic key (excluding Xantolis), diagnostic character combinations for all genera, and the necessary taxonomic combinations for Pichonia, Planchonella, Pleioluma, and Sersalisia to render each genus monophyletic.

Phylogenetic studies favour the unification of Pennisetum, Cenchrus and Odontelytrum (Poaceae): a combined nuclear, plastid and morphological analysis, and nomenclatural combinations in Cenchrus.

Twenty-five genera having sterile inflorescence branches were recognized as the bristle clade within the x = 9 Paniceae (Panicoideae). Within the bristle clade, taxonomic circumscription of Cenchrus (20-25 species), Pennisetum (80-140) and the monotypic Odontelytrum is still unclear. Several criteria have been applied to characterize Cenchrus and Pennisetum, but none of these has proved satisfactory as the diagnostic characters, such as fusion of bristles in the inflorescences, show continuous variation. A phylogenetic analysis based on morphological, plastid (trnL-F, ndhF) and nuclear (knotted) data is presented for a representative species sampling of the genera. All analyses were conducted under parsimony, using heuristic searches with TBR branch swapping. Branch support was assessed with parsimony jackknifing. Based on plastid and morphological data, Pennisetum, Cenchrus and Odontelytrum were supported as a monophyletic group: the PCO clade. Only one section of Pennisetum (Brevivalvula) was supported as monophyletic. The position of P. lanatum differed among data partitions, although the combined plastid and morphology and nuclear analyses showed this species to be a member of the PCO clade. The basic chromosome number x = 9 was found to be plesiomorphic, and x = 5, 7, 8, 10 and 17 were derived states. The nuclear phylogenetic analysis revealed a reticulate pattern of relationships among Pennisetum and Cenchrus, suggesting that there are at least three different genomes. Because apomixis can be transferred among species through hybridization, its history most likely reflects crossing relationships, rather than multiple independent appearances. Due to the consistency between the present results and different phylogenetic hypotheses (including morphological, developmental and multilocus approaches), and the high support found for the PCO clade, also including the type species of the three genera, we propose unification of Pennisetum, Cenchrus and Odontelytrum. Species of Pennisetum and Odontelytrum are here transferred into Cenchrus, which has priority. Sixty-six new combinations are made here.

Species delimitation and evolution of metal bioaccumulation in the lichenized Acarospora smaragdula (Ascomycota, Fungi) complex.

The crustose lichenized fungi in the Acarosporaceae are splendid examples of organisms managing to survive in extremely harsh environments, such as highly mineralized rocks and low-pH habitats. Some representatives of the Acarospora smaragdula complex are known to accumulate substantial amounts of potentially toxic metals including iron and copper, resulting in populations with highly divergent coloration and morphology. These populations have often been treated as distinct species by lichen taxonomists. Parsimony and parsimony jackknifing analyses of β-tubulin, nuclear ITS rDNA, and mtSSU rDNA sequence data sets was used to investigate the evolution of iron and copper accumulation and the production of the secondary compound norstictic acid in populations within the A.smaragdula species complex in Sweden, with additional samples mainly from Norway and the UK. Phylogenetic species recognition (concordance of single-gene phylogenies) was used to investigate species delimitations. Seven species are recognized in the complex. Atypically green, copper-accumulating samples, often given species rank, do not form a distinct group but are nested within A.smaragdula s. str., indicating that this ability is widespread in this species. Rust-coloured, iron-accumulating samples form two well supported separate groups, indicating that two morphologically distinct, obligate, iron-accumulating species are present, but facultatively iron-accumulating populations occur in at least one additional species. Norstictic acid, sometimes claimed to characterize the whole A.smaragdula complex, is only present in A.smaragdula s. str. The evolutionary significance of metal accumulation in Acarospora is discussed, as is the significance of our results for the application of phylogenetic species recognition/gene tree concordance-based species recognition, and DNA barcoding. © The Willi Hennig Society 2009.

DNA taxonomy of Swedish Catenulida (Platyhelminthes) and a phylogenetic framework for catenulid classification

Specimens of Catenulida were collected at 34 localities in Sweden. We used 18S rDNA, 28S rDNA, ITS-5.8S, and cytochrome oxidase I (COI) nucleotide sequences to infer phylogeny from parsimony jackknifing and Bayesian analysis. Our dataset contained 74 ingroup terminals and 5111 characters. The results show a basal split between a clade consisting of the marine Retronectidae+the limnic Catenulidae, and a second clade consisting of the limnic Stenostomidae. The hypothesis of the marine Retronectidae as the sister group of the limnic Catenulida is rejected. The recently introduced genus Anokkostenostomum Noreña, Damborenea & Brusa, 2005 results as non-monophyletic, and Suomina Marcus, 1945 as a group inside Catenula Dugès, 1832. Therefore, we propose to render Anokkostenostomum a new junior synonym of Stenostomum Schmidt, 1848, and Suomina a new junior synonym of Catenula. Consequently, the new combinations Catenula evelinae (Marcus, 1945), Catenula sawayai (Marcus, 1945), and Catenula turgida (Zacharias, 1902) are proposed, and 14 species are returned to their original genus, Stenostomum. The molecular phylogenetic hypothesis is used to identify and discriminate catenulid species. In our material, we found 12 species of Catenulida new to Sweden, and four species new to science, all of which are distinguishable by morphological characters.

Multi-gene phylogeny of the pantropical subfamily Chrysophylloideae (Sapotaceae): evidence of generic polyphyly and extensive morphological homoplasy.

We present a molecular phylogeny of 26 out of the 28 currently accepted genera in the subfamily Chrysophylloideae (Sapotaceae) using parsimony, parsimony jackknifing, and Bayesian inference. A data matrix of 8984 characters was obtained from DNA sequences of seven chloroplast loci, two nuclear loci, indels coded as binary characters, and morphology. Our phylogenetic reconstruction suggests that Chrysophyllum, Pouteria, and Pradosia, as well as some sections within Chrysophyllum and Pouteria, are all polyphyletic. These taxa were previously described largely on the basis of unique combinations of states for a set of morphological characters. Mapping some of these characters onto one of the most parsimonious trees indicates that the symplesiomorphic flower in the subfamily was probably 5-merous, had stamens inserted in the tube orifice, staminodes, seeds with foliaceous cotyledons, exserted radicle, and endosperm. These characters have subsequently been lost multiple times and cannot be used as synapomorphies to support broad generic concepts. Despite the high degree of homoplasy some well-defined clades can be described on the basis of alternative character state combinations. Also, many of these well-supported clades appear to be restricted to particular geographical areas (e.g. all taxa in Australasia form a monophyletic group). Hence, we suggest that the segregate genera Aningeria, Malacantha, and Martiusella may ultimately be resurrected, and probably also Donella and Gambeya, but their circumscriptions are still unclear. One species, Chrysophyllum cuneifolium, may have originated from a hybridization event between continents where the maternal genome (cpDNA) comes from South America and the nuclear genome comes from Africa. © The Willi Hennig Society 2008.

Molecular phylogeny ofPlanchonella(Sapotaceae) and eight new species from New Caledonia

We present a hypothesis of phylogenetic relationships inPlanchonella(Sapotaceae, Chrysophylloideae) using nrDNA (ITS) data analysed with parsimony jackknifing and Bayesian inference. Results from these two approaches yield similar majority‐rule consensus trees.Planchonella(formerly included inPouteria) is a monophyletic genus of trees and shrubs with 60 species currently recognised. They are distributed in Australia (13 spp.), New Caledonia (32 spp.), the Pacific islands (10 spp.), and New Guinea (5 spp.), but more molecular research is needed in order to identify all members in New Guinea and Malesia. Three strongly supported clades and several subclades were recovered by our analysis. Two clades are restricted to New Caledonia. Within these are eight recently discovered species that are described here (P. crenata, P. glauca, P. latihila, P. luteocostata, P. mandjeliana, P. povilana, P. roseoloba, P. rufocostata), each with a conservation assessment. Our molecular phylogeny shows that some of these taxa, initially believed to be conspecific or closely related, represent separate lineages that deserve species rank. Two of these species are restricted to areas with near‐future mining activities, which is why urgent conservation assessments are needed. Several subspecies or varieties ofP. myrsinifoliaandP. cotinifoliaare recognised in Australia. Neither of these two species is monophyletic in its present circumscription, and for this reasonP. cotinifoliavar.pubescensis given species rank here asP. pubescens. We also amend the generic description ofPlanchonella, a genus best distinguished on a character combination of stamens positioned just below (rarely in) the tube orifice, a multi‐seeded fruit, and foliaceous cotyledons embedded in endosperm.

Filling a gap in the phylogeny of flatworms: relationships within the Rhabdocoela (Platyhelminthes), inferred from 18S ribosomal DNA sequences

The phylogeny of the Rhabdocoela, a species‐rich taxon of free‐living flatworms, is reconstructed based on complete 18S rDNA sequences. The analysis includes 62 rhabdocoels and 102 representatives of all major flatworm taxa. In total, 46 new sequences are used, 41 of them from rhabdocoel species, five from proseriates. Phylogenetic analysis was performed using maximum parsimony and Bayesian inference. Clade support was evaluated with parsimony jackknifing, Bremer support indices and Bayesian posterior probabilities. The resulting cladogram corroborates that the Rhabdocoela is monophyletic, but its sister group remains uncertain. The ‘Dalyellioida’ and the ‘Typhloplanoida’, both former rhabdocoel subtaxa, are polyphyletic. Within the Rhabdocoela the monophyletic Kalyptorhynchia, characterized by a muscular proboscis, forms the sister group of all other rhabdocoels. The Schizorhynchia is a monophyletic subtaxon of the Kalyptorhynchia, with the split proboscis as a synapomorphy. Except for the Dalyelliidae and the Typhloplanidae, both freshwater taxa, none of the ‘families’ previously included in the ‘Typhloplanoida’ and the ‘Dalyellioida’ appears to be monophyletic. As a result of this analysis, three existing and four new taxon names are formally defined following the rules of the Phylocode.

Generic delimitations in the family Stictidaceae (Ostropales, Ascomycota): the Stictis–Conotrema problem

The family Stictidaceae (Ostropales, Ascomycota) contains both lichenized and non-lichenized fungi. Here, we test if Conotrema (lichenized) and Stictis (non-lichenized) as currently delimited are distinct monophyletic genera, by parsimony and parsimony jackknifing analyses of combined nuclear rDNA (ITS and partial LSU rDNA) and mitochondrial SSU rDNA sequence data matrices. The study includes four species of Stictis, three species of Conotrema, and representatives of the related Schizoxylon (lichenized), Odontotrema, Carestiella (at least sometimes associated with algae), Cryptodiscus and Thelotrema (lichenized). In all analyses, the Conotrema species were nested within Stictis with high support. Thus, we conclude that Conotrema are only lichenized representatives of Stictis. The type species of the two generic names, C. urceolatum and S. radiata, are sister taxa in our analyses. Furthermore, the analysis gave no support for the present infrageneric classification of Stictis. Carestiella socia (the type of Carestiella) and the two representatives of Schizoxylon studied were also nested within Stictis s. lat. The Odontotremataceae is the sister group to the Stictidaceae, and Cryptodiscus foveolaris groups with Thelotrema rather than with the Stictidaceae. We conclude that lichenization in the Stictidaceae does not characterize natural groups, and that Conotrema should be considered a synonym to Stictis, as predicted by anatomical characteristics. The new combinations Stictis urceolatum and Stictis populorum are made.

Phylogenetic analysis of bovine pestiviruses: testing the evolution of clinical symptoms.

This study presents a phylogenetic analysis of 115 bovine pestiviruses. A sequence data set from the 5' untranslated genomic region was analyzed with maximum parsimony, bootstrapping and parsimony jackknifing. We tested for the proposed classifications of the group and analyzed the evolution of the symptoms associated with Pestivirus infections in bovines. Based on the historical framework provided by our phylogenetic trees, we also characterized the extent and importance of contamination caused in biologicals by the virus. Our phylogenetic analyses showed that the previously defined genotypes are monophyletic, except for genotype 1a. Based on our cladograms, we propose the existence of more than 12 monophyletic groups within the species BVDV 1. The mapping of clinical symptoms suggests that the emergence of some genotypes could have been driven by a change in the pathogenic process. Enteric Problems appear to be ancestral, while Reproductive and Respiratory Problems arise with the emergence of genotypes 1b, 1d and the herein-proposed genotype Arg 1. The distribution of contaminant strains on the cladograms shows that pestiviral contamination is a common process, and also suggests that a contaminated product might be a vehicle for virus dispersion. Implications for virus evolution, virus taxonomy, veterinary medicine and biotechnology are discussed.

Phylogeny and Biogeography of Dionysia (Primulaceae)

Cladistic analyses using parsimony jackknifing and Bayesian inference were performed to elucidate phylogenetic relationships in the genus Dionysia. DNA sequences from one nuclear DNA region (ITS) and from two chloroplast genome regions (rps16 intron and trnL‐trnF) were used with and without the addition of a morphological character matrix. A total of 43 species from Primulaceae were sequenced, including 39 species of Dionysia. The results strongly suggest that the sister species to the rest of genus Dionysia are Dionysia balsamea and D. hissarica, not D. mira. No section and subsection with more than one species of Dionysia as currently circumscribed appears to be monophyletic. Instead, species often belong to clades corresponding to geographical proximity. Earlier proposed notions of evolutionary trends and geographical migrations in Dionysia are discussed.

The phylogenetic relationships of the cyanobacterial lichens in the Lecanorales suborder Peltigerineae.

We present major cladistic analyses of the Lecanoromycetes (Ascomycota, Fungi) focusing on the Lecanorales suborder Peltigerineae, a group including the majority of the cyanobacterial lichens. DNA sequence datasets from the mtSSU and nLSU rDNA were produced and analyzed with maximum parsimony and parsimony jackknifing. The results suggest that the Lecanorales is monophyletic. The Peltigerineae (including Placynthiaceae, Peltigeraceae, Lobariaceae, Nephromataceae, Collemataceae, Coccocarpiaceae, Pannariaceae, and Massalongia) is likewise a monophyletic group. The Lobariaceae, and Lobaria in the traditional sense, are strongly supported as monophyletic, in contrast to results of other investigations based on nITS rDNA data. Pseudocyphellaria may be paraphyletic. Placynthiaceae is the sister group to the Collemataceae and Collema may be nested within Leptogium. Pannariaceae in the traditional sense is not a monophyletic group. Finally, the Lecanorineae is nonmonophyletic in all analyses, and the Cladoniineae and Teloschistineae are nested within the Lecanorineae in the combined analysis.

The phylogenetic relationships of the cyanobacterial lichens in the Lecanorales suborder Peltigerineae

We present major cladistic analyses of the Lecanoromycetes (Ascomycota, Fungi) focusing on the Lecanorales suborder Peltigerineae, a group including the majority of the cyanobacterial lichens. DNA sequence datasets from the mtSSU and nLSU rDNA were produced and analyzed with maximum parsimony and parsimony jackknifing. The results suggest that the Lecanorales is monophyletic. The Peltigerineae (including Placynthiaceae, Peltigeraceae, Lobariaceae, Nephromataceae, Collemataceae, Coccocarpiaceae, Pannariaceae, and Massalongia) is likewise a monophyletic group. The Lobariaceae, and Lobaria in the traditional sense, are strongly supported as monophyletic, in contrast to results of other investigations based on nITS rDNA data. Pseudocyphellaria may be paraphyletic. Placynthiaceae is the sister group to the Collemataceae and Collema may be nested within Leptogium. Pannariaceae in the traditional sense is not a monophyletic group. Finally, the Lecanorineae is nonmonophyletic in all analyses, and the Cladoniineae and Teloschistineae are nested within the Lecanorineae in the combined analysis.

A phylogenetic perspective on larval spine morphology in Leucorrhinia (Odonata: Libellulidae) based on ITS1, 5.8S, and ITS2 rDNA sequences

Leucorrhinia (Odonata, Anisoptera, Libellulidae) consists of 14–15 species with a holarctic distribution. We have combined the morphological characters of a previous study with sequence data from the ITS1, 5.8S rDNA, and ITS2 regions of the nuclear ribosomal repeat. Cloning was used to investigate the intra-individual variation and such variation was found in all investigated species. Parsimony jackknifing was used to identify supported groups. The effect of sequence alignment and gap coding was explored by a modified sensitivity analysis. Loss of spines in Leucorrhinia larvae has occurred twice: once in Europe and once in North America. The role of spines as a defence against predation is discussed in a phylogenetic context.

Evolutionary relationships within the protostome phylum Sipuncula: a molecular analysis of ribosomal genes and histone H3 sequence data

The phylogenetic relationships of the members of the phylum Sipuncula are investigated by means of DNA sequence data from three nuclear markers, two ribosomal genes (18S rRNA and the D3 expansion fragment of 28S rRNA), and one protein-coding gene, histone H3. Phylogenetic analysis via direct optimization of DNA sequence data using parsimony as optimality criterion is executed for 12 combinations of parameter sets accounting for different indel costs and transversion/transition cost ratios in a sensitivity analysis framework. Alternative outgroup analyses are also performed to test whether they affected rooting of the sipunculan topology. Nodal support is measured by parsimony jackknifing and Bremer support values. Results from the different partitions are highly congruent, and the combined analysis for the parameter set that minimizes overall incongruence supports monophyly of Sipuncula, but nonmonophyly of several higher taxa recognized for the phylum. Mostly responsible for this is the split of the family Sipunculidae in three main lineages, with the genus Sipunculus being the sister group to the remaining sipunculans, the genus Phascolopsis nesting within the Golfingiiformes, and the genus Siphonosoma being associated to the Phascolosomatidea. Other interesting results are the position of Phascolion within Golfingiidae and the position of Antillesoma within Aspidosiphonidae. These results are not affected by the loci selected or by the outgroup chosen. The position of Apionsoma is discussed, although more data would be needed to better ascertain its phylogenetic affinities. Monophyly of the genera with multiple representatives ( Themiste, Aspidosiphon, and Phascolosoma) is well supported, but not the monophyly of the genera Nephasoma or Golfingia. Interesting phylogeographic questions arise from analysis of multiple representatives of a few species.

Phylogenetic hypotheses: Cladoniaceae, Stereocaulaceae, Baeomycetaceae, and Icmadophilaceae revisited

Parsimony analyses of SSU rDNA sequences were conducted to examine phylogenetic relationships of selected genera within the families Cladoniaceae, Stereocaulaceae, Icmadophilaceae and Baeomycetaceae (lichen-forming ascomycetes). The analyses included 93 taxa (84 species) representing various groups of ascomycetous fungi. Analyses of the matrix with pre-aligned sequences were performed using heuristic and parsimony ratchet searches, and support values for the same matrix were calculated using parsimony jackknifing. The results support the recognition of the four families. Cladoniaceae are recircumscribed to accommodate Cladia, Cladonia, Heterodea, Metus, Pilophorus, Pycnothelia, Ramalea, Thysanothecium and the newly erected genus Carassea. Myelorrhiza is excluded from the family, while the status of other potential members, Calathaspis, Gymnoderma s.str. and Squamella, remains unresolved. Baeomycetaceae include Baeomyces and Phyllobaeis. Stereocaulaceae include Stereocaulon only, although the status of Muhria is still unclear. Finally, Icmadophilaceae include Dibaeis, Endocena, Knightiella, Icmadophila, Siphula and Thamnolia, while the status of Pseudobaeomyces and Siphulella requires further elucidation. The genus Cladonia appeared to be a polyphyletic assemblage, and accordingly, a new genus Carassea S. Stenroos, gen. nov., represented by C. connexa (Vain.) S. Stenroos, comb. nov., is described. Carassea is most closely related to Pycnothelia and Metus in the Cladoniaceae. Siphula, represented in the present analysis by six species, is not monophyletic, and is in need of reclassification.

A cladistic analysis of Scinax (Anura: Hylidae)

The hylid frog genus Scinax ranges from southern Mexico to eastern Argentina. The 84 recognized species are currently arranged in five groups: the Scinax rubra, Scinax rostrata, Scinax catharinae, Scinax staufferi, and Scinax perpusilla species groups. Although a few species groups are distinctive, compelling evidence of monophyly is lacking for three groups, and their interrelationships remain virtually unknown. With the objective of testing the monophyly of the species groups and exploring their interrelationships, a cladistic analysis was performed, employing 86 characters from osteology, myology, adult and larval external morphology, reproductive biology, and karyotypes taken from 36 representative species of the five species groups plus eight outgroup taxa. The analysis of the resulting data matrix shows moderately well-supported (as assessed through Bremer support and parsimony jackknifing) S. catharinae and S. rostrata species groups. The S. staufferi species group is polyphyletic, as currently composed, given that some of its species are nested within the S. catharinae species group. The only representative of the S. perpusilla species group is the sister species of the S. catharinae species group. Although weakly supported, the representatives of the S. rostrata and S. staufferi species groups are nested, separately, within the S. rubra species group. In order to have a taxonomy consistent with these results, it is proposed not to recognize the S. rubra group but to recognize a more inclusive rubra clade that includes all studied exemplars of the former S. rubra group, plus some of the S. staufferi group and all of the S. rostrata group. Similarly, a catharinae clade is recognized, within which all studied exemplars of the S. catharinae group (that now includes Scinax berthae and Scinax agilis) and the one studied representative of the S. perpusilla group are included.

A cladistic analysis of Scinax (Anura: Hylidae).

The hylid frog genus Scinax ranges from southern Mexico to eastern Argentina. The 84 recognized species are currently arranged in five groups: the Scinax rubra, Scinax rostrata, Scinax catharinae, Scinax staufferi, and Scinax perpusilla species groups. Although a few species groups are distinctive, compelling evidence of monophyly is lacking for three groups, and their interrelationships remain virtually unknown. With the objective of testing the monophyly of the species groups and exploring their interrelationships, a cladistic analysis was performed, employing 86 characters from osteology, myology, adult and larval external morphology, reproductive biology, and karyotypes taken from 36 representative species of the five species groups plus eight outgroup taxa. The analysis of the resulting data matrix shows moderately well-supported (as assessed through Bremer support and parsimony jackknifing) S. catharinae and S. rostrata species groups. The S. staufferi species group is polyphyletic, as currently composed, given that some of its species are nested within the S. catharinae species group. The only representative of the S. perpusilla species group is the sister species of the S. catharinae species group. Although weakly supported, the representatives of the S. rostrata and S. staufferi species groups are nested, separately, within the S. rubra species group. In order to have a taxonomy consistent with these results, it is proposed not to recognize the S. rubra group but to recognize a more inclusive rubra clade that includes all studied exemplars of the former S. rubra group, plus some of the S. staufferi group and all of the S. rostrata group. Similarly, a catharinae clade is recognized, within which all studied exemplars of the S. catharinae group (that now includes Scinax berthae and Scinax agilis) and the one studied representative of the S. perpusilla group are included.

New genes for phylogenetic studies of lichenized fungi: glyceraldehyde-3-phosphate dehydrogenase and beta-tubulin genes

Abstract:Primers for amplification and sequencing of partial glyceraldehyde-3-phosphate dehydrogenase (gpd) gene were designed for lichenized fungi. The 5′ gpd primer is most probably fungal specific, since a BLAST search in GenBank found identical sequences only from ascomycetous taxa, whereas the 3′ gpd primer was more universal. Utility of the gpd primers and previously designed beta-tubulin primers was tested in nine lichen taxa. Both the gpd and beta-tubulin primer pairs amplified in most of the taxa examined: the gpd primers generated a c.1100 nucleotide fragment, whereas the PCR product obtained from the beta-tubulin primers was c.900 nucleotides long. The gpd amplification products of Cladonia arbuscula and C. rangiferina were sequenced and both were found to contain three introns, the length of which varied between 49 to 83 nucleotides. To examine the applicability of gpd sequences in resolving relationships within Ascomycota, trees were calculated from 22 fungal gpd sequences obtained from GenBank together with the twoCladonia sequences using parsimony jackknifing. The gpd tree was compared with the SSU rDNA tree of the respective species (or genera). A similar analysis of the beta-tubulin gene was not performed, because only a few beta-tublin sequences from the same taxa were available in GenBank. The gpd tree was well resolved but in conflict with the SSU rDNA tree. In contrast to the SSU rDNA tree, the gpd tree did not support the monophyly of the Ascomycota. Analysis of the combined data set produced a tree very similar to that of the SSU rDNA data. However, the relationship of Lecanorales to the other orders remained unresolved. Even though gpd and beta-tubulin are highly conserved proteins, the third codon positions and introns are variable and both genes have the potential for inferring phylogenetic relationships at the lower taxonomic levels in the lichenized fungi. The two genes may be useful even below species level, depending on the species investigated.