New insights into Paleozoic charophyte morphology and phylogeny

Examination of Paleozoic charophyte fructifications using microscopy and high-resolution x-ray synchrotron microtomography has revealed that most of them have a utricle that forms a supplementary calcified cover around the gyrogonite. All Paleozoic families with utricles have been assigned to the Sycidiales. We consider the Moellerinaceae to occupy a central position in the phylogeny of the Charophyta. From these, one group of descendants constitutes the gyrogonites inside the utricles of the Sycidiales; a second descendant goup is thought to be the utricle-free ancestors of the Charales prior to inversion of spiralization and reduction in cell number. The Sycidiales have a multilayered wall and an internal vesicle, but their families are distinguished by diversity in orientation of external cells, complexity of the utricle wall, and in presence or absence of antheridia. The solidly packed structure of the utricle is believed to be an organ protecting the zygote against dessication. We interpret the morphological similarities between Paleozoic Sycidiales and Mesozoic Clavatoraceae, both with a utricle, as homoplasous rather than expressing a true phylogenetic relationship. We confirm that some umbellids might correspond to utricles of charophytes.     

Paleoamerican morphology in the context of European and East Asian late Pleistocene variation: implications for human dispersion into the New World

Early American crania show a different morphological pattern from the one shared by late Native Americans. Although the origin of the diachronic morphological diversity seen on the continents is still debated, the distinct morphology of early Americans is well documented and widely dispersed. This morphology has been described extensively for South America, where larger samples are available. Here we test the hypotheses that the morphology of Early Americans results from retention of the morphological pattern of Late Pleistocene modern humans and that the occupation of the New World precedes the morphological differentiation that gave rise to recent Eurasian and American morphology. We compare Early American samples with European Upper Paleolithic skulls, the East Asian Zhoukoudian Upper Cave specimens and a series of 20 modern human reference crania. Canonical Analysis and Minimum Spanning Tree were used to assess the morphological affinities among the series, while Mantel and Dow-Cheverud tests based on Mahalanobis Squared Distances were used to test different evolutionary scenarios. Our results show strong morphological affinities among the early series irrespective of geographical origin, which together with the matrix analyses results favor the scenario of a late morphological differentiation of modern humans. We conclude that the geographic differentiation of modern human morphology is a late phenomenon that occurred after the initial settlement of the Americas.     

DNA amplification: New insights into its mechanism

DNA amplification is a process whereby a limited part of the genome is increased in copy number with various consequences for the cell. It is frequently observed in cancer cells and it can be induced in mammalian cells grown in culture as well as in tumor cells when these are subjected to growth inhibiting drugs. In recent years new insights into the mechanisms involved in DNA amplification have been obtained; discussion of these will form the major subject of this short review.     

Amorphophallus: New insights into pollen morphology and the chemical nature of the pollen wall

In pollen characters, Amorphophallus is one of the most diverse genera in the Araceae. The present work is a critical survey of contradicting reports on the impact of acetolysis treatment on Amorphophallus pollen, on the chemical nature of the outer pollen wall layer and of electron-dense (dark) granules found within it. Furthermore, we wanted to clarify the pollen polarity and to test conclusions based on different preparation techniques. Pollen morphology of 25 species is investigated by light microscopy, scanning electron microscopy and transmission electron microscopy. Our results show that Amorphophallus pollen is not resistant to acetolysis treatment. The use of different transmission electron microscopy staining methods proved the polysaccharide nature of the outer pollen wall layer and of the granules within it. Moreover, an additional thin surface layer was found in all investigated species. Microspores in early and late tetrad stages show that the less convex side of the microspore is the proximal face and the more convex side the distal face. The extrusion of pollen in strands is illustrated for the first time by light microscopy and scanning electron microscopy. Furthermore, observations of pollen in water showed that in some of the investigated species the pollen wall is shed immediately before pollen tube formation.     

Leaf morphology and anatomy of Manilkara Adans. (Sapotaceae) from northeastern Brazil

Aiming to complement the group’s taxonomy, the objective of this work is to describe and characterise, searching for morpho-anatomical patterns, the leaf anatomy of the Manilkara species occurring in the northeast region of Brazil. The leaves of all Manilkara species were analysed, using herbaria specimens and collections made in the field by the authors. The characterisation of the leaf blades was performed according to the usual protocols in vegetal anatomy. As diagnosed by anatomical characters, the Manilkara Adans. genus presents petioles with a thick cuticle, a uniseriate epidermis, a sclerenchymatous sheath involving the vascular bundle, and laticifers on the cortical and medullar regions. The anatomical data increase the M. elata and M. huberi synonymisation and show the importance of the anatomy as a complementary tool to the taxonomy based on external morphology.     

Investigations into the late Pleistocene and Holocene history of vegetation and climate in Santa Catarina (S Brazil)

Palynological studies have been carried out on three highland peat bogs, and one situated on the Atlantic coastal plain. In the highlands, the late Pleistocene (14,000 - 10,000 uncal B.P.) vegetation was dominated by campos (grassland). Scattered stands of Araucaria forests were preserved in deep valleys. In the region of the sites at Morro da Igreja and Serra do Rio Rastro, the dominance of campos vegetation continued until about 1000 B.P. while at the Serra da Boa Vista site there was an expansion of Atlantic pluvial forest elements followed by Araucaria forests at the beginning of the Holocene. A general expansion of A. angustifolia, clearly related to a change towards an increasingly moist climate, can be dated to the present millenium. On the coastal plains, the late Pleistocene vegetation was dominated by Myrtaceae which were replaced by tropical taxa in the Holocene. The lowland profile (Poço Grande) also covers part of the upper Holocene, where the rich flora of the Atlantic pluvial forests can be characterized by taxa including Alchornea, Urticales and Rapanea. Close to the coring site, there was a repeated alternation between two different dune communities (4840 - 4590 B.P.), followed by a lake stage with aquatic plant succession (4590 - 4265 B.P.), plant communities dominated by Rapanea (4265 - 4230 B.P.) and the spread of Alchornea (4230 - 3525 B.P.). Late Pleistocene climate conditions (14,000 - 10,000 B.P.) can be described as cold and relatively dry, possibly including an equivalent of the Younger Dryas period. In the Holocene, there were changes from a warm and drier climate (10,000 -3000 B.P.) to a cool and more moist regime (ca. 3000 -ca. 1000 B.P.) and finally to a cool and very moist period (from around 1000 B.P.).     

New insights into the systematics of the enigmatic Polynesian sandpipers Aechmorhynchus parvirostris and Prosobonia leucoptera

With only a single extant representative, endemic to the Tuamotu Archipelago, the Polynesian sandpipers (Aechmorhynchus and Prosobonia) may have had a larger distribution in Eastern Polynesia in the past, with four endemic taxa. Although these aberrant sandpipers' membership to the Scolocapidae has been well supported, finding their closest living taxa has proved difficult and the phylogenetic relationships of these taxa have remained unresolved. We present the first molecular analysis of the Polynesian sandpipers, including sampling of the only known specimen of the extinct Prosobonia leucoptera, collected in 1773. Based on mitochondrial and nuclear gene sequence data, the phylogenetic analyses demonstrate that the Polynesian sandpipers were sister taxa and belonged to the clade that included the other sandpipers (Calidris and allies) and turnstones (Arenaria), although without a close relative among extant genera. Divergence time estimates suggested that the lineage leading to Prosobonia diverged from the other extant sandpipers during the Oligocene and that either the Line Islands or the Tuamotu Archipelago were probably the first archipelagos colonized by the Prosobonia lineage. On the basis of these results, we suggest that Aechmorhynchus parvirostris and Prosobonia leucoptera be regarded as related species within the same genus, and thus that the senior name Prosobonia be used for both taxa.     

New insights on Early Pleistocene Nyctereutes from the Balkans based on material from Dafnero-3 (Greece) and Varshets (Bulgaria)

The study examines the taxonomy of the newly excavated Nyctereutes material from Dafnero-3 (Greece) and the unpublished specimens from Varshets (Bulgaria). One cranium and a hemimandible from Dafnero-3 site are ascribed to the species Nyctereutes megamastoides, already known from Dafnero-1 and other Early Pleistocene sites of Greece. Two additional crania and a hemimandible from Dafnero-3, along with the Varshets dental material are attributed to N. tingi. This species was previously reported in the Early Pliocene Megalo Emvolon fauna of northern Greece but the record has been challenged and a review is provided. The new findings of Dafnero-3, and Varshets, extend the presence of N. tingi in southeastern Europe until the beginning of the middle Villafranchian and confirm the geographic expansion of the taxon towards West. Moreover, the co-existence of N. tingi and N. megamastoides in Dafnero-3 is reported for the first time in Europe. A paleoecological analysis based on the lower molar method agrees with the existing hypothesis of a more omnivorous N. megamastoides and a more carnivorous N. tingi ecomorph, which further justifies their local co-existence. Additionally, the quantitative analyses of N. tingi from Dafnero-3, and Varshets reveal a possible succession from N. tingi to N. vulpinus, as already supported by previous studies.     

New insights into the pollen morphology of the genus Gunnera (Gunneraceae)

Pollen grains from ten species of Gunnera, chosen to represent the six different subgenera in the genus, were examined using light and scanning electron microscopy. The aim of the study was to explore characters that have the potential to define different types of pollen within Gunnera and to study the evolution of these characters in light of the phylogeny of the genus. According to our results, there are three main types of pollen in the examined species of Gunnera. Type 1, unique for the South American species G. herteri (subgenus Ostenigunnera), is characterised by an imperfect reticulum with sinuous undulating-creasted muri. A reticulum with equidimensional polygonal lumina is typical for the plesiomorphic type of pollen (type 2) present in subgenera Gunnera, Misandra and Panke. Lastly, pollen grains of subgenera Pseudogunnera and Milligania are characterised by a reticulum with lumina of variable shape and size (type 3). In G. macrophylla (subgenus Pseudogunnera), the lumina in the apocolpia are of a different shape and size from the lumina in the mesocolpia (type 3a), while in G. dentata, G. monoica and G. cordifolia (subgenus Milligania), the lumina are identical in the apocolpium and the mesocolpium (type 3b).

The identification of pollen types will possibly allow the interpretation of the different specimens of Tricolpites reticulatus, the fossil species believed to be allied to the extant Gunnera.

In addition to the revision on the pollen of Gunnera, a brief comparison between the pollen of this genus and its sister group Myrothamnus is reported.     

New insights into the systematics of North Atlantic Gaidropsarus (Gadiformes,Gadidae): flagging synonymies and hidden diversity

Gaidropsarus Rafinesque, 1810 is a genus of marine fishes, commonly known as rocklings, comprising 14 living species and showing a high ecological diversity from the intertidal zone to the deep sea. The systematics of this group has been controversial due to a general lack of representative specimens and the conservative morphology exhibited. A multidisciplinary approach combining the analysis of meristic data and the DNA barcode standard was applied in a species delimitation approach. Individuals representing eight valid and three unnamed species were collected, morphologically identified and archived in several museum collections. Comparison of DNA sequences shows complex results, furthering the idea of the difficult identification of specimens based on traditional taxonomy. DNA barcoding supports synonymies, like G. biscayensis–G. macrophthalmus and G. guttatus–G. mediterraneus, agreeing with the extensive overlaps observed in the meristic variables analysed and suggesting a reduction in the number of species. Genetic distances showed pairs of closely related species like G. granti–G. vulgaris and G. argentatus–G. ensis, the latter being only distinguished by one main distinctive character. Four deep-water specimens, morphologically classified only to the genus level, constituted three independent taxa apart from the ones present in this study and with no barcode matches in the repository databases. They could represent new records for the North Atlantic or unknown species of this genus. The results obtained show that more studies will be necessary to solve the systematics of this branch of the Gadiformes.     

New insights into the functional morphology of the male copulatory apparatus of bullid gastropods

In this study, semi-thin sections stained with histochemical techniques and transmission electron microscopy were used to obtain new data about the morphology and function of the male copulatory apparatus of the cephalaspidean gastropod Bulla striata. The apparatus comprises a vestibule, a penial papilla and a prostate consisting of a coiled unbranched tube ending in a blind caecum. The penial papilla and the coiled tubular prostate are enclosed by a muscular sheath, which is continuous with the muscular tissue of the vestibule. The epithelium lining the lumen of the vestibule is formed by ciliated and mucus-secreting cells. Two new types of subepithelial secretory cells were discovered in this region. The penial papilla is a muscular structure without secretory cells in the epithelium lining the narrow lumen. The tubule that constitutes the prostate possesses a muscular wall and can be divided in three distinct regions: a non-secretory duct connected to the penial papilla, a glandular region rich in large secretory cells and the terminal caecum containing just a few small secretory cells. In the terminal blind caecum, the muscular sheath is fused with the muscular wall of the tubular prostate. Large numbers of spermatozoa were found in the glandular region and in the terminal caecum of the prostate. A new functional mechanism is proposed to explain penial eversion during copulation. This differs from a previous hypothesis in two main aspects: (1) existence of a permanent penial papilla in mature animals acting as a functional penis and (2) functional role of vestibule during copulation, which everts and surrounds the penial papilla, while the latter protrudes outwards.     

Neodiptera: New insights into the adult morphology and higher level phylogeny of Diptera (Insecta)

This paper outlines several aspects of the skeleto-muscular organization of the adult prothorax and cervix pertaining to the ground pattern of Diptera, which in turn leads to the characterization of Neodiptera, a higher level dipterous taxon which includes Brachycera and bibionomorph Nematocera ( sensu Hennig). The monophyly of Neodiptera is firmly supported by four skeleto-muscular modifications of the pronoto-cervical region. The bibionomorph Nematocera are shown to be paraphyletic in terms of Brachycera. On more preliminary evidence it is argued that the fundamental dichotomy of the extant Diptera lies between a 'polyneuran' clade which includes Trichoceridae, Tipuloidea, Tanyderidae, and Ptychopteridae and an 'oligoneuran' clade which includes all the remaining groups. Preliminary evidence for a sister group relationship between Blephariceroidea and Culicomorpha is also provided. The possible adaptational significance of the cervical specializations in Neodiptera is discussed.     

New insights into pollen morphology and its implications in the phylogeny of Sanguisorba L. (Rosaceae; Sanguisorbeae)

Pollen morphology of 13 species of the genus Sanguisorba (Rosaceae) was examined by light and scanning electron microscopy. The pollen morphology divided the genus into two main groups: (A) tricolporate with tapered colpus tips, and (B) either tricolporate with opened colpus tip or hexacolporate. The former group was further subdivided into those without vestibulum (A1: S.?alpina, S.?dodecandra, and S.?filiformis) and those with vestibulum (A2: S.?agrimonoides, S.?ancestroides, S.?annua, S.?cretica, S.?minor, and S.?verrucosa), whereas the latter group was subdivided into those with colpus narrow and similar to the A1 type (B1: S.?canadensis and S.?diandra), those with colpus and mesocolpus somewhat equally wide and forming hexacolporate aperture (B3: S.?albiflora, S.?armena, S.?media, S.?menziesii, S.?parviflora, S.?stipulata, and S.?tenuifolia), and those with colpus intermediate between the B1 and B3 types (B2: S.?hakusanensis, S.?microcephala, S.?obtusa, S.?officinalis, S.?polygama, and S.?sitchensis). It is suggested that the A1 type aperture would have evolved to the A2 type as a specialized form and the B types (B1 to B3) in a direction in which the hexacolporate aperture was derived. Implications of pollen morphology for infrageneric classification of Sanguisorba are discussed and the results are compared with molecular phylogenetic studies.     

Tiny treefrogs in the Pleistocene: Phylogeography of Dendropsophus oliveirai in the Atlantic Forest and associated enclaves in northeastern Brazil

Wet and dry cyclical periods through the Pleistocene have been suggested as drivers of isolation and, consequently, divergence and expansions of populations associated with forest environments in the Neotropics. In northeastern Brazil, forests patches restricted to high altitudes led to the formation of regionally distinct ecosystems, known as “Atlantic Forest enclaves” or “brejos de altitude.” Herein, we investigated the population history of the treefrog Dendropsophus oliveirai, a species associated with the Atlantic Forest and areas of enclaves in the Northeast region of the Brazil. We obtained sequences for one mitochondrial and two nuclear loci of 50 samples from 16 localities to investigate the population structure and demographic history across northeastern Atlantic Forest enclaves. Our data revealed that D. oliveirai exhibits three mtDNA haplogroups: low and highlands above the São Francisco River; low and highlands between Chapada Diamantina enclaves and northern populations of coastal Bahia and Sergipe States; lowlands below the Contas River. Divergence times estimates revealed that population splits occurred during the Pleistocene. Our data recovered low genetic differentiation between enclaves and lowland populations north and south of the São Francisco River, which supports a scenario of recent contact and repeated recolonization of enclaves by lowland populations during the humid Pleistocene phases. Species tree, and temporal and spatial population structures may indicate that D. oliveirai is actually a species complex.     

Welwitschiaceae from the Lower Cretaceous of northeastern Brazil

Welwitschiaceae, a family in the Gnetales, is known today from only one extant species, Welwitschia mirabilis. This species is distributed in the Namibian desert, along the western coast of southern Africa, about 10 km inland from the coast. Very little is known about the fossil record of this family. Lower Cretaceous megafossils of various organs, assigned to Welwitschiaceae, are presented here. These fossils include young stems with paired cotyledons attached (Welwitschiella austroamericana n. gen. et sp.), isolated leaves (Welwitschiophyllum brasiliense n. gen. et sp.), and axes bearing male cones (Welwitschiostrobus murili n. gen. et sp.). They were collected in the Crato Formation, which is dated by palynomorphs and ostracods as Late Aptian (114 to 112 million years ago). These sediments are exposed in the Araripe Basin of northeastern Brazil. This study brings together new information of the megafossil record of Welwitschia-like plants and also reports of pollen said to be similar to that of Welwitschia from Lower Cretaceous sediments.     

DNA barcoding of coastal ichthyofauna from Bahia,northeastern Brazil,South Atlantic: High efficiency for systematics and identification of cryptic diversity

In spite of the utility of DNA barcoding in discriminating marine fish species, few studies using this technique are available along the Brazilian coast, which encompasses nearly 9000 km, distinct biogeographic regions and endemism rates. Therefore, in the present work, we provided the first DNA barcode library based on sequencing of the mitochondrial gene cytochrome c oxidase subunit I (COI) for Actinopterygii fish from northeastern coast of Brazil. The database included 459 specimens from 54 genera, 31 families and 13 orders, with mean interspecific genetic distance of and mean intraspecific distance of 0.15%. As a result, the neighbor-joining (NJ) tree resulted in the molecular identification of 78 taxa. Cryptic species were indicated in Eucinostomus (mojarras) since the 82 specimens morphologically identified as Eucinostomus melanopterus or Eucinostomus sp. formed five clusters with intraspecific genetic distances up to 0.31% and lowest interspecific distance of 11.14%. Eucinostomus jonesii and Eucinostomus harengulus were identified for the first time in ichthyofauna inventories from northeastern Brazil. The present work has also added new records to the Barcode of Life Data System (BOLD), thereby favoring the identification of other specimens at different life stages or in forensic studies. The present dataset reinforces the reliability of DNA barcoding to estimate the overlooked richness of marine fish in Brazilian coast with application in conservation, ecology, systematics and fisheries.