Divisestylus gen. nov. (aff. Iteaceae), a fossil saxifrage from the Late Cretaceous of New Jersey, USA

Fossilized flowers and fruits from the Upper Cretaceous (Turonian, ca. 90 million years [my] before present) Raritan Formation of New Jersey are described as the new genus Divisestylus with two species, D. brevistamineus and D. longistamineus. The fossils are fusainized and three-dimensionally preserved. Morphological characteristics suggest affinities with extant Saxifragaceae and Iteaceae, two closely related families in Saxifragales. Similarities include a pentamerous perianth, calyx fused below into a hypanthium with free sepal lobes above, haplostemonous androecium with stamens situated opposite the calyx lobes, inferior ovary, bicarpellate gynoecium, numerous ovules on axile placentas, conspicuous intrastaminal nectary ring, and capsulate fruit opening apically. The unique fusion of the gynoecium, with carpels and stigmas fused but styles free, indicates closer affinities with extant Iteaceae, whereas other characters, such as basifixed anthers in D. brevistamineus, tricolpate and striate pollen grains, and anomocytic stomata, indicate closer affinities to Saxifragaceae. Cladistic analyses utilizing molecular data from a previously published analysis and morphological data as well as morphological data alone demonstrate the fossils share a more recent common ancestor with Iteaceae than Saxifragaceae, thereby making Divisestylus the oldest fossils known with clear affinities to Iteaceae.     

Morphological, biochemical and molecular characterization of Herpetomonas samuelpessoai camargoi n. subsp., a trypanosomatid isolated from the flower of the squash Cucurbita moschata

We report the morphological, biochemical and molecular characteristics of a trypanosomatid isolated from the flower of Cucurbita moschata. Although the trypanosomatid was isolated from a plant, the lack of recognition of Phytomonas-specific molecular markers based on spliced-leader and ribosomal genes as well as by monoclonal antibodies specific for Phytomonas argues against assigning it to this genus. Because the isolate displayed typical opisthomastigote forms in culture, it is assigned to the genus Herpetomonas. Analysis of randomly amplified polymorphic DNA (RAPD) patterns and characterization of ribosomal SSU and ITS markers suggest that it is more closely related to H. samuelpessoai than to any other species. However, the presence of spined flagellates in culture (displaying lateral expansions of the plasma membrane originating near the flagellar pocket) and isolate-specific RAPD fingerprints argue strongly that the trypanosomatid belongs to a new subspecies, for which the name Herpetomonas samuelpessoai camargoi n. subsp. is proposed.     

From one to ten in a single stroke--resolving the European Eumidasanguinea (Phyllodocidae, Annelida) species complex

We investigate the genetic structure of European lineages of the polychaete Eumida sanguinea with mitochondrial COI and nuclear ITS, and demonstrate the presence of ten cryptic species. Within the E. sanguinea species complex there are six different white pigmentation patterns but only three of these are unique for a single species. No other consistent morphological differences were observed. We give new species names to seven of the lineages from our study, providing examples of combined morphological and molecular diagnoses, apply the available name E. sanguinea to one species, and leave two lineages with single representatives unnamed. We also include new data and a diagnosis for the poorly known E. notata, which also belongs to the E. sanguinea species complex.     

Tsukubamonas globosa n. gen., n. sp., a novel excavate flagellate possibly holding a key for the early evolution in "Discoba"

We report the ultrastructure and phylogenetic position of a free-living heterotrophic flagellate, Tsukubamonas globosa n. gen., n. sp. This flagellate was isolated from a pond in the University of Tsukuba, Japan. Under light microscopy, the spherical vegetative cells were naked and highly vacuolated, and always swam with rotating motion. Electron microscopic observations revealed that T. globosa possessed a ventral feeding groove, which is one of the hallmark characteristics of the supergroup Excavata. The position of T. globosa was unresolved in the small subunit ribosomal RNA phylogeny. On the other hand, a multigene phylogeny using α-tubulin, β-tubulin, actin, heat shock protein 90, and translation elongation factor 2 robustly united T. globosa with members of the "Discoba" clade of Excavata, composed of jakobids, euglenozoans, and heteroloboseans, although the precise position of T. globosa in this clade remained unresolved. Our detailed morphological comparisons elucidated that T. globosa possessed a novel set of morphological features, and could not be classified into any taxa in the Discoba clade. Instead we classified T. globosa into Tsukubamonadidae n. fam. under Tsukubamonadida n. ord.     

Development and molecular characterisation of the microsporidian Schroedera airthreyi n. sp. in a freshwater bryozoan Plumatella sp. (Bryozoa: Phylactolaemata)

The development of a new species of microsporidian, infecting a freshwater Plumatellid bryozoan, is described. The small-subunit rDNA, internal transcribed spacer region (ITS), and partial large-subunit rDNA genes were sequenced. Phylogenetic analysis demonstrated that the parasite clustered with Schroedera plumatellae. However, while there were morphological affinities with this species, significant differences were also observed. The infection initially appeared as a roughening of the peritoneum lining the metacoelom of the bryozoan. This roughening resolved into meront-infected syncytia, composed of interconnected cells of the body wall that detached to float in the coleomic cavity. Spores were observed to develop within these syncytia. All stages of development were diplokaryotic in contrast to S. plumatellae, which has a distinct monokaryotic merogony preceding sporogony. The infection was pathogenic to the host. Direct bryozoan-bryozoan transmission was not observed. We propose to name the microsporidian Schroedera aithreyi n. sp.     

Correct identification of species makes the amoebozoan rRNA tree congruent with morphology for the order Leptomyxida Page 1987; with description of Acramoeba dendroida n. g., n. sp., originally misidentified as 'Gephyramoeba sp.'

Morphological identification of protists remains an expert task, especially for little known and poorly described species. Culture collections normally accept organisms under the name provided by depositors and are not responsible for identification. Uncritical acceptance of these names by molecular phylogeneticists may result in serious errors of interpretation of phylogenetic trees based on DNA sequences, making them appear more incongruent with morphology than they really are. Several cases of misidentification in a major culture collection have recently been reported. Here we provide evidence for misidentifications of two more gymnamoebae. The first concerns “Gephyramoeba sp.” ATCC 50654; it is not Gephyramoeba, a leptomyxid with lobose pseudopods, but a hitherto undescribed branching amoeba with fine, filamentous subpseudopods named here Acramoeba dendroida gen. et sp. nov. We also sequenced 18S rRNA of Page's strain of Rhizamoeba saxonica (CCAP 1570/2) and show that it is the most deeply branching leptomyxid and is not phylogenetically close to ‘Rhizamoeba saxonica’ ATCC 50742, which was misidentified. Correcting these misidentifications improves the congruence between morphological diversity of Amoebozoa and their rRNA-based phylogenies, both for Leptomyxida and for the Acramoeba part of the tree. On morphological grounds we transfer Gephyramoebidae from Varipodida back to Leptomyxida and remove Flamella from Leptomyxida; sequences are needed to confirm these two revisions.     

Description and Phylogenetic Relationships of Spumochlamys perforata n. sp. and Spumochlamys bryora n. sp. (Amoebozoa, Arcellinida)

ABSTRACT. Spumochlamys perforata n. sp. and Spumochlamys bryora n. sp. were isolated and described from dry epiphytic moss. The morphology and ultrastructure of both species clearly demonstrate that they belong to the genus Spumochlamys (family Microchlamyiidae). They differ from its only described member, Spumochlamys iliensis (as well as from species of Microchlamys ), in the relief of the dorsal surface of the test, revealed by scanning electron microscopy, which can represent a good characteristic for species identification. They also differ in the structure of the dorsal part of the test wall (especially S. perforata ). Small subunit ribosomal DNA-based molecular phylogenetic analyses show that Spumochlamys is a deeply branching lineage of the Arcellinida, without any close affinities. Actin gene sequence analysis places this genus within the Tubulinea, close to two other arcellinid lineages but without forming a monophyletic group with them. These data together strongly suggest that the lack of resolution in the arcellinid molecular phylogenies is due to serious undersampling of taxa, a limited number of sequence data, and high divergence rates in most of the species.     

Mataza hastifera n. g., n. sp.: a possible new lineage in the Thecofilosea (Cercozoa)

A new cercozoan flagellate Mataza hastifera n. g., n. sp. is described from a surface seawater sample collected in Tokyo Bay. Cells are 3-5 μm in diameter and have two flagella. The cells alternate between swimming and stationary states in culture. Swimming cells have a nodding motion. Phylogenetic analyses using small subunit rDNA sequences demonstrate that M. hastifera belongs to the clade comprised of only environmental sequences closely related to thecofilosean cercozoans. Ultrastructural observations reveal that M. hastifera is quite similar to members of Cryomonadida, an order in Thecofilosea, and especially to Cryothecomonas spp. The cell of M. hastifera is covered with a thin double-layered theca and possesses a cylinder-shaped extrusome, as reported from cryomonads. On the other hand, the funnel that is characteristic of cryomonads was not found in the flagellar pit of M. hastifera. Combining both morphological and molecular analyses, we conclude that M. hastifera is a new lineage in Thecofilosea and suggest that Thecofilosea may be a larger group than previously thought.     

Design principles for active transport systems

We have previously described simple models for active transport and have derived steady state equations for the unidirectional flux of substrate in terms of a minimal set of kinetic parameters. Here we consider how to maximize the pumping rate of a carrier-enzyme through the optimal utilization of the ATP hydrolysis reaction. The equations for net flux contain rate constants and dissociation constants and these determine the maximum velocities and affinities measured in transport kinetic analysis. It is assumed that the rate constants can evolve to the diffusion limited rate of substrate binding as has apparently occurred in the enzyme triosephosphate isomerase (Knowles & Albery, 1977). The dissociation constants of the rate limiting intermediates fit the affinities for substrates on different sides of the membrane and are dependent on the basic free energy levels (Hill, 1976) of the carrier substrate system. From our analysis it is clear that there are three ways to design a system with optimal affinities and that the choice is linked to the sequence of substrate binding. It is possible to use free energy differences of isomerization (Boyer, 1975) or ligand-ligand interactions (Weber, 1975) both of which have been described previously, but which are incorporated here into a unified treatment. A third possibility is to couple the binding step of a transported ligand to the progress of a chemical reaction as might occur, for example, if Na⁺ must be bound before the carrier can be phosphorylated. In this way the free energy of hydrolysis can be used not only to drive the overall pumping reaction, but also to fix differentially the affinity for substrate on either side of the membrane, as required for rapid pumping.     

Evaluating microevolutionary models for the early settlement of the New World: the importance of recurrent gene flow with Asia

Different scenarios attempting to describe the initial phases of the human dispersal from Asia into the New World have been proposed during the last two decades. However, some aspects concerning the population affinities among early and modern Asians and Native Americans remain controversial. Specifically, contradictory views based mainly on partial evidence such as skull morphology or molecular genetics have led to hypotheses such as the "Two Waves/Components" and "Single Wave" or "Out of Beringia" model, respectively. Alternatively, an integrative scenario considering both morphological and molecular variation has been proposed and named as the "Recurrent Gene Flow" hypothesis. This scenario considers a single origin for all the Native Americans, and local, within-continent evolution plus the persistence of contact among Circum-Arctic groups. Here we analyze 2D geometric morphometric data to evaluate the associations between observed craniometric distance matrix and different geographic design matrices reflecting distinct scenarios for the peopling of the New World using basic and partial Mantel tests. Additionally, we calculated the rate of morphological differentiation between Early and Late American samples under the different settlement scenarios and compared our findings to the predicted morphological differentiation under neutral conditions. Also, we incorporated in our analyses some variants of the classical Single Wave and Two Waves models as well as the Recurrent Gene Flow model. Our results suggest a better explanatory performance of the Recurrent Gene Flow model, and provide additional insights concerning affinities among Asian and Native American Circum-Arctic groups.     

Phylogeny and biogeography of the "Australian" trichostomes (Ciliophora: Litostomata)

The phylogenetic relationships of members of the ciliate class Litostomatea were determined by a molecular phylogeny using the small subunit of the ribosomal RNA (ssu-rRNA) gene and a morphological phylogeny based on ultrastructural analyses of the group. Molecular analyses consistently supported the monophyly of Trichostomatia, Entodiniomorphida and the "Australian" trichostomes but provided limited support for a monophyletic Vestibuliferida and Haptoria. The results of the morphological analyses depended on the way in which the dataset was treated: "unordered" and "ordered" recovered a monophyletic Trichostomatia, Haptoria and the "Australian" trichostomes but challenged the monophyly of Entodinimorphida and Vestibuliferida; "dollo" recovered a monophyletic Trichostomatia and Entodiniomorphida but at the cost of a greatly longer tree than either "unordered" or "ordered" datasets. The monophyly of each "Australian" trichostome family was supported in all analyses and by both approaches. These results suggest that the trichostome ciliates may have become associated with mammals in Gondwana with the "Australian" trichostome ciliates entering Australia with primitive herbivorous marsupials. Subsequent diversification of the "Australian" families was probably a result of dietary specialization and oral and cortical synapomorphies define each family. We decline at this time to erect a formal taxon name for the "Australian" trichostomes due to the instability of other superfamilial taxa within the Litosomatea and concerns about the stability of tree topology until a better taxon sample of litostome ciliates is available.     

Investigations on the sheathed bacteriumHaliscomenobacter hydrossis gen.n., sp.n., isolated from activated sludge

The morphological and physiological properties of a recently isolated, thin, sheathed bacterium are described. The observations demonstrate that this bacterium differs from the known sheathed microorganisms in many characteristics. The following nomenclature is therefore proposed:Haliscomenobacter hydrossis gen.n. sp.n. The growth of this bacterium in activated sludge is discussed.     

The novel marine gliding zooflagellate genus Mantamonas (Mantamonadida ord. n.: Apusozoa)

Mantamonasis a novel genus of marine gliding zooflagellates probably related to apusomonad and planomonad Apusozoa. Using phase and differential interference contrast microscopy we describe the type species Mantamonas plasticasp. n. from coastal sediment in Cumbria, England. Cells are ～5μm long, ～5μm wide, asymmetric, flattened, biciliate, and somewhat plastic. The posterior cilium, on which they glide smoothly over the substratum, is long and highly acronematic. The much thinner, shorter, and almost immobile anterior cilium points forward to the cell's left. These morphological and behavioural traits suggest thatMantamonasis a member of the protozoan phylum Apusozoa. Analyses of 18S and 28S rRNA gene sequences of Mantamonas plasticaand a second genetically very different marine species from coastal sediment in Tanzania show Mantamonasas a robustly monophyletic clade, that is very divergent from all other eukaryotes. 18S rRNA trees mostly placeMantamonaswithin unikonts (opisthokonts, Apusozoa, and Amoebozoa) but its precise position varies with phylogenetic algorithm and/or taxon and nucleotide position sampling; it may group equally weakly as sister to Planomonadida, Apusomonadida or Breviata. On 28S rRNA and joint 18/28S rRNA phylogenies (including 11 other newly obtained apusozoan/amoebozoan 28S rRNA sequences) it consistently strongly groups with Apusomonadida (Apusozoa).     

A likelihood method for detecting trait-dependent shifts in the rate of molecular evolution

Rate heterogeneity within groups of organisms is known to exist even when closely related taxa are examined. A wide variety of phylogenetic and dating methods have been developed that aim either to test for the existence of rate variation or to correct for its bias. However, none of the existing methods track the evolution of features that account for observed rate heterogeneity. Here, we present a likelihood model that assumes that rate variation is caused, in part, by species' intrinsic characteristics, such as a particular life-history trait, morphological feature, or habitat association. The model combines models of sequence and character state evolution such that rates of sequence change depend on the character state of a lineage at each point in time. We test, using simulations, the power and accuracy of the model to determine whether rates of molecular evolution depend on a particular character state and demonstrate its utility using an empirical example with halophilic and freshwater daphniids.     

Relaxed clocks and inferences of heterogeneous patterns of nucleotide substitution and divergence time estimates across whales and dolphins (Mammalia: Cetacea)

Various nucleotide substitution models have been developed to accommodate among lineage rate heterogeneity, thereby relaxing the assumptions of the strict molecular clock. Recently developed "uncorrelated relaxed clock" and "random local clock" (RLC) models allow decoupling of nucleotide substitution rates between descendant lineages and are thus predicted to perform better in the presence of lineage-specific rate heterogeneity. However, it is uncertain how these models perform in the presence of punctuated shifts in substitution rate, especially between closely related clades. Using cetaceans (whales and dolphins) as a case study, we test the performance of these two substitution models in estimating both molecular rates and divergence times in the presence of substantial lineage-specific rate heterogeneity. Our RLC analyses of whole mitochondrial genome alignments find evidence for up to ten clade-specific nucleotide substitution rate shifts in cetaceans. We provide evidence that in the uncorrelated relaxed clock framework, a punctuated shift in the rate of molecular evolution within a subclade results in posterior rate estimates that are either misled or intermediate between the disparate rate classes present in baleen and toothed whales. Using simulations, we demonstrate abrupt changes in rate isolated to one or a few lineages in the phylogeny can mislead rate and age estimation, even when the node of interest is calibrated. We further demonstrate how increasing prior age uncertainty can bias rate and age estimates, even while the 95% highest posterior density around age estimates decreases; in other words, increased precision for an inaccurate estimate. We interpret the use of external calibrations in divergence time studies in light of these results, suggesting that rate shifts at deep time scales may mislead inferences of absolute molecular rates and ages.     

Molecular Phylogenetic Positions and Ultrastructure of Marine Gregarines (Apicomplexa) Cuspisella ishikariensis n. gen., n. sp. and Loxomorpha cf. harmothoe from Western Pacific scaleworms (Polynoidae)

Marine gregarines are unicellular parasites of invertebrates commonly found infecting the intestine and coelomic spaces of their hosts. Situated at the base of the apicomplexan tree, marine gregarines offer an opportunity to explore the earliest stages of apicomplexan evolution. Classification of marine gregarines is often based on the morphological traits of the conspicuous feeding stages (trophozoites) in combination with host affiliation and molecular phylogenetic data. Morphological characters of other life stages such as the spore are also used to inform taxonomy when such stages can be found. The reconstruction of gregarine evolutionary history is challenging, due to high levels of intraspecific variation of morphological characters combined with relatively few traits that are taxonomically unambiguous. The current study combined morphological data with a phylogenetic analysis of small subunit rDNA sequences to describe and establish a new genus and species (Cuspisella ishikariensis n. gen., n. sp.) of marine gregarine isolated from the intestine of a polynoid host (Lepidonotus helotypus) collected from Hokkaido, Japan. This new species possesses a set of unusual morphological traits including a spiked attachment apparatus and sits on a long branch on the molecular phylogeny. Furthermore, this study establishes a molecular phylogenetic position for Loxomorpha cf. harmothoe, a previously described marine gregarine, and reveals a new group of gregarines that infect polynoid hosts.     

End of an enigma: <Emphasis Type="Italic">Aenigmopteris</Emphasis> belongs in <Emphasis Type="Italic">Tectaria</Emphasis> (Tectariaceae: Polypodiopsida)

The phylogenetic affinities of the fern genus Aenigmopteris have been the subject of considerable disagreement, but until now, no molecular data were available from the genus. Based on the analysis of three chloroplast DNA regions (rbcL, rps16-matK, and trnL-F) we demonstrate that Aenigmopteris dubia (the type species of the genus) and A. elegans are closely related and deeply imbedded in Tectaria. The other three species of genus are morphologically very similar; we therefore transfer all five known species into Tectaria. Detailed morphological comparison further shows that previously proposed diagnostic characters of Aenigmopteris fall within the range of variation of a broadly circumscribed Tectaria.     

<Emphasis Type="Italic">Saccharothrix tharensis</Emphasis> sp. nov., an actinobacterium isolated from the Thar Desert,India

The taxonomic provenance of a filamentous actinobacterial strain isolated from a desert soil was established using a polyphasic approach. The strain has chemotaxonomic and morphological properties consistent with its classification in the genus Saccharothrix. It forms a distinct branch in the Saccharothrix 16S rRNA gene tree, related to the type strain of Saccharothrix saharensis (96.7%) but was distinguished readily from it using a combination of phenotypic properties. The genotypic and phenotypic data show that the strain represents a novel species in the genus Saccharothrix, for which the name Saccharothrix tharensis sp. nov. is proposed with the type strain TD-093^T (=?KCTC 39724^T?=?MCC 2832^T).