Análise da composição e distribuição geográfica dos atuns da costa brasileira (Perciformes: Scombridae: Thunnini)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Developmental basis of limb homology in Pleurodiran turtles, and the identity of the hooked element in the chelonian tarsus

Although Pleurodiran turtles represent an important component of extant turtle radiation, our knowledge of the development and homology of limb bones in turtles rests mostly upon observations made on derived members of the Cryptodiran clade. Herein, we describe limb development in three pleurodirans: Podocnemis unifilis Troschel, 1848, Podocnemis sextuberculata Cornalia, 1849 and Phrynops hilarii (Dumeril and Bibron, 1835), in an effort to contribute to filling this anatomical gap. For earlier stages of limb development, we described the Y-shaped condensation that gave rise to the zeugopodial cartilages, and differentiation of the primary axis/digital arch that reveals the invariant pattern common to tetrapods. There are up to four central cartilaginous foci in the carpus, and the proximal tarsale is formed by the fusion of the fibulare, intermedium, and centrale 4. Digital development is similar for the five digits. Changes in toe V occur predominantly in the distal tarsale 5. Ontogenetic reduction of phalanges is observed in toe V of Podocnemis. Based on these results, we suggest that the hooked element present in the chelonian tarsus, and traditionally recognized as a modified fifth metatarsale, is actually the fifth distal tarsale. Additionally, our data on limb development of pleurodiran turtles supply more taxonomically comprehensive information to interpret limb configuration within the chelonian clade. (C) 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 155, 845-866.

A new genus and species of inseminating characid fish from the rio Xingu basin (Characiformes: Characidae)

Phallobrycon adenacanthus, is described as a new genus and species belonging to Clade A characids of Malabarba & Weitzman (2003). The new taxon is the only characid possessing two developed spines on unbranched portions of fifth, sixth and seventh anal-fin rays associated with intumescent glandular tissue on the anterior portion of the anal fin of sexually mature males. Other non-exclusive diagnostic features of the new genus (observed in male specimens) are: urogenital papilla modified into a copulatory organ, absence of pelvic-fin hooks and glandular tissue not organized into an organ. The presence of these features in members of Clade A and other characids is discussed in order to hypothesize the relationships of Phallobrycon.

Gastrotricha: A Marine Sister for a Freshwater Puzzle

Background: Within an evolutionary framework of Gastrotricha Marinellina flagellata and Redudasys fornerise bear special interest, as they are the only Macrodasyida that inhabit freshwater ecosystems. Notwithstanding, these rare animals are poorly known; found only once (Austria and Brazil), they are currently systematised as incertae sedis. Here we report on the rediscovery of Redudasys fornerise, provide an account on morphological novelties and present a hypothesis on its phylogenetic relationship based on molecular data. Methodology/Principal Findings: Specimens were surveyed using DIC microscopy and SEM, and used to obtain the 18 S rRNA gene sequence; molecular data was analyzed cladistically in conjunction with data from 42 additional species belonging to the near complete Macrodasyida taxonomic spectrum. Morphological analysis, while providing new information on taxonomically relevant traits (adhesive tubes, protonephridia and sensorial bristles), failed to detect elements of the male system, thus stressing the parthenogenetic nature of the Brazilian species. Phylogenetic analysis, carried out with ML, MP and Bayesian approaches, yielded topologies with strong nodal support and highly congruent with each other. Among the supported groups is the previously undocumented clade showing the alliance between Redudasys fornerise and Dactylopodola agadasys; other strongly sustained clades include the densely sampled families Thaumastodermatidae and Turbanellidae and most genera. Conclusions/Significance: A reconsideration of the morphological traits of Dactylopodola agadasys in light of the new information on Redudasys fornerise makes the alliance between these two taxa very likely. As a result, we create Anandrodasys gen. nov. to contain members of the previously described D. agadasys and erect Redudasyidae fam. nov. to reflect this novel relationship between Anandrodasys and Redudasys. From an ecological perspective, the derived position of Redudasys, which is deeply nested within the Macrodasyida clade, unequivocally demonstrates that invasion of freshwater by gastrotrichs has taken place at least twice, in contrast with the single event hypothesis recently put forward.

Molecular phylogeny and morphometric analyses reveal deep divergence between Amazonia and Atlantic Forest species of Dendrophryniscus

Dendrophryniscus is an early diverging clade of bufonids represented by few small-bodied species distributed in Amazonia and the Atlantic Forest. We used mitochondrial (414 bp of 12S, 575 bp of 16S genes) and nuclear DNA (785 bp of RAG-1) to investigate phylogenetic relationships and the timing of diversification within the genus. These molecular data were gathered from 23 specimens from 19 populations, including eight out of the 10 nominal species of the genus as well as Rhinella boulengeri. Analyses also included sequences of representatives of 18 other bufonid genera that were publically available. We also examined morphological characters to analyze differences within Dendrophryniscus. We found deep genetic divergence between an Amazonian and an Atlantic Forest clade, dating back to Eocene. Morphological data corroborate this distinction. We thus propose to assign the Amazonian species to a new genus, Amazonella. The species currently named R. boulengeri, which has been previously assigned to the genus Rhamphophryne, is shown to be closely related to Dendrophryniscus species. Our findings illustrate cryptic trends in bufonid morphological evolution, and point to a deep history of persistence and diversification within the Amazonian and Atlantic rainforests. We discuss our results in light of available paleoecological data and the biogeographic patterns observed in other similarly distributed groups. (C) 2011 Elsevier Inc. All rights reserved.

A Novel Stress-Induced Sugarcane Gene Confers Tolerance to Drought, Salt and Oxidative Stress in Transgenic Tobacco Plants

Background: Drought is a major abiotic stress that affects crop productivity worldwide. Sugarcane can withstand periods of water scarcity during the final stage of culm maturation, during which sucrose accumulation occurs. Meanwhile, prolonged periods of drought can cause severe plant losses. Methodology/Principal Findings: In a previous study, we evaluated the transcriptome of drought-stressed plants to better understand sugarcane responses to drought. Among the up-regulated genes was Scdr1 (sugarcane drought-responsive 1). The aim of the research reported here was to characterize this gene. Scdr1 encodes a putative protein containing 248 amino acids with a large number of proline (19%) and cysteine (13%) residues. Phylogenetic analysis showed that ScDR1is in a clade with homologs from other monocotyledonous plants, separate from those of dicotyledonous plants. The expression of Scdr1 in different varieties of sugarcane plants has not shown a clear association with drought tolerance. Conclusions/Significance: The overexpression of Scdr1 in transgenic tobacco plants increased their tolerance to drought, salinity and oxidative stress, as demonstrated by increased photosynthesis, water content, biomass, germination rate, chlorophyll content and reduced accumulation of ROS. Physiological parameters, such as transpiration rate (E), net photosynthesis (A), stomatal conductance (gs) and internal leaf CO2 concentration, were less affected by abiotic stresses in transgenic Scdr1 plants compared with wild-type plants. Overall, our results indicated that Scdr1 conferred tolerance to multiple abiotic stresses, highlighting the potential of this gene for biotechnological applications.

An additional basal sauropodomorph specimen from the Upper Triassic Caturrita Formation, southern Brazil, with comments on the biogeography of plateosaurids

We describe an additional saurischian specimen from the Caturrita Formation (Norian) of the Parana Basin, southern Brazil. This material was collected in the 1950s and remained unstudied due to its fragmentary condition. Detailed comparisons with other saurischians worldwide reveal that some characters of the ilium, including the low ventral projection of the medial wall of the acetabulum and its concave ventral margin, together with the short triangular shape of the pre-acetabular process and its mound-like dorsocaudal edge, resemble those of sauropodomorphs such as Plateosaurus and Riojasaurus. This set of traits suggests that MN 1326-V has affinities with basal Sauropodomorpha, probably closer to plateosaurians than to Saturnalia-like taxa. Previous records of this clade in the Caturrita Formation include Unaysaurus, which has been related to Plateosaurus within Plateosauridae. Alternative schemes suggest that plateosaurids include Plateosaurus plus the Argentinean 'prosauropods' Coloradisaurus and Riojasaurus. Both hypotheses raise biogeographic questions, as a close relationship between faunas from South America and Europe excluding Africa and North America is not supported by geological and biostratigraphical evidence. Additionally, the absence of plateosaurids in other continents suggests that the geographical distribution of this taxon is inconsistent with the geological history of western Pangaea, and this demands further investigations of the phylogeny of sauropodomorphs or improved sampling.

The skull of the Upper Cretaceous snake Dinilysia patagonica Smith-Woodward, 1901, and its phylogenetic position revisited

The cranial anatomy of Dinilysia patagonica, a terrestrial snake from the Upper Cretaceous of Argentina, is redescribed and illustrated, based on high-resolution X-ray computed tomography and better preparations made on previously known specimens, including the holotype. Previously unreported characters reinforce the intriguing mosaic nature of the skull of Dinilysia, with a suite of plesiomorphic and apomorphic characters with respect to extant snakes. Newly recognized plesiomorphies are the absence of the medial vertical flange of the nasal, lateral position of the prefrontal, lizard-like contact between vomer and palatine, floor of the recessus scalae tympani formed by the basioccipital, posterolateral corners of the basisphenoid strongly ventrolaterally projected, and absence of a medial parietal pillar separating the telencephalon and mesencephalon, amongst others. We also reinterpreted the structures forming the otic region of Dinilysia, confirming the presence of a crista circumfenestralis, which represents an important derived ophidian synapomorphy. Both plesiomorphic and apomorphic traits of Dinilysia are treated in detail and illustrated accordingly. Results of a phylogenetic analysis support a basal position of Dinilysia, as the sister-taxon to all extant snakes. The fossil taxa Yurlunggur, Haasiophis, Eupodophis, Pachyrhachis, and Wonambi appear as derived snakes nested within the extant clade Alethinophidia, as stem-taxa to the crown-clade Macrostomata. The hypothesis of a sister-group relationship between Dinilysia and Najash rionegrina, as suggested by some authors, is rejected by the results of our analysis.

A comparative transcriptome analysis reveals expression profiles conserved across three Eimeria spp. of domestic fowl and associated with multiple developmental stages

Coccidiosis of the domestic fowl is a worldwide disease caused by seven species of protozoan parasites of the genus Eimeria. The genome of the model species, Eimeria tenella, presents a complexity of 55-60 MB distributed in 14 chromosomes. Relatively few studies have been undertaken to unravel the complexity of the transcriptome of Eimeria parasites. We report here the generation of more than 45,000 open reading frame expressed sequence tag (ORESTES) cDNA reads of E. tenella, Eimeria maxima and Eimeria acervulina, covering several developmental stages: unsporulated oocysts, sporoblastic oocysts, sporulated oocysts, sporozoites and second generation merozoites. All reads were assembled to constitute gene indices and submitted to a comprehensive functional annotation pipeline. In the case of E. tenella, we also incorporated publicly available ESTs to generate an integrated body of information. Orthology analyses have identified genes conserved across different apicomplexan parasites, as well as genes restricted to the genus Eimeria. Digital expression profiles obtained from ORESTES/EST countings, submitted to clustering analyses, revealed a high conservation pattern across the three Eimeria spp. Distance trees showed that unsporulated and sporoblastic oocysts constitute a distinct clade in all species, with sporulated oocysts forming a more external branch. This latter stage also shows a close relationship with sporozoites, whereas first and second generation merozoites are more closely related to each other than to sporozoites. The profiles were unambiguously associated with the distinct developmental stages and strongly correlated with the order of the stages in the parasite life cycle. Finally, we present The Eimeria Transcript Database (http://www.coccidia.icb.usp.br/eimeriatdb), a website that provides open access to all sequencing data, annotation and comparative analysis. We expect this repository to represent a useful resource to the Eimeria scientific community, helping to define potential candidates for the development of new strategies to control coccidiosis of the domestic fowl. (C) 2011 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Revisiting the Acanthamoeba species that form star-shaped cysts (genotypes T7, T8, T9, and T17): characterization of seven new Brazilian environmental isolates and phylogenetic inferences

Free-living amoebae of the genus Acanthamoeba are the agents of both opportunistic and non-opportunistic infections and are frequently isolated from the environment. Of the 17 genotypes (T1-T17) identified thus far, 4 (T7, T8, T9, and T17) accommodate the rarely investigated species of morphological group I, those that form large, star-shaped cysts. We report the isolation and characterization of 7 new Brazilian environmental Acanthamoeba isolates, all assigned to group I. Phylogenetic analyses based on partial (similar to 1200 bp) SSU rRNA gene sequences placed the new isolates in the robustly supported clade composed of the species of morphological group I. One of the Brazilian isolates is closely related to A. comandoni (genotype T9), while the other 6, together with 2 isolates recently assigned to genotype T17, form a homogeneous, well-supported group (2-0% sequence divergence) that likely represents a new Acanthamoeba species. Thermotolerance, osmotolerance, and cytophatic effects, features often associated with pathogenic potential, were also examined. The results indicated that all 7 Brazilian isolates grow at temperatures up to 40 degrees C, and resist under hvperosmotic conditions. Additionally, media conditioned by each of the new Acanthamoeba isolates induced the disruption of SIRC and HeLa cell monolayers.

Morphological and molecular evidence demonstrates the amphi-Atlantic distribution of Laurencia catarinensis (Ceramiales, Rhodophyta)

We performed morphological and molecular studies of Laurencia catarinensis from the Canary Islands. This species has an entangled habit, cushion-like tuft formation, cortical cell walls, slightly to markedly projecting near the apex, and lacking lenticular thickenings in medullary cells. We inferred its phylogenetic position by analyzing the chloroplast-encoded rbcL gene sequences from 41 samples. The results demonstrate that specimens of L. catarinensis from the Canary Islands, where it is referred to as L. intricata, and those from Brazil (including specimens from the type locality in Santa Catarina) form a monophyletic clade with low genetic divergence (0-0.9%). In contrast, specimens of L. intricata from the type locality in Cuba, Mexico, and the USA were clearly distinct from L. catarinensis collected in Brazil and the Canary Islands, as shown by high genetic divergence values (4.9-5.7%). The type material of L. catarinensis from Brazil allowed us to identify all samples from the Canarian Archipelago as L. catarinensis. These findings expand the known geographical distribution of L. catarinensis to the eastern Atlantic Ocean and demonstrate an amphi-Atlantic distribution of the species.

Streptomyces brevispora sp nov and Streptomyces laculatispora sp nov., actinomycetes isolated from soil

The taxonomic positions of two actinomycetes isolated from a hay meadow soil sample were determined using a polyphasic approach. The isolates had chemical and morphological properties typical of streptomycetes and formed a distinct 16S rRNA gene subclade together with the type strain Streptomyces drozdowiczii NRRL B-24297(T). DNA DNA relatedness studies showed that the three strains belonged to different genomic species. The organisms were also distinguished using a combination of phenotypic properties. On the basis of these data it is proposed that the isolates be assigned to the genus Streptomyces as Streptomyces brevispora sp. nov. and Streptomyces laculatispora sp. nov., with BK160(T) (=KACC 21093(T) =NCIMB 14702(T)) and BK166(T) (=KACC 20907(T) =NCIMB 14703(T)) as the respective type strains.

Diversity and Physiological Characterization of D-Xylose-Fermenting Yeasts Isolated from the Brazilian Amazonian Forest

Background: This study is the first to investigate the Brazilian Amazonian Forest to identify new D-xylose-fermenting yeasts that might potentially be used in the production of ethanol from sugarcane bagasse hemicellulosic hydrolysates. Methodology/Principal Findings: A total of 224 yeast strains were isolated from rotting wood samples collected in two Amazonian forest reserve sites. These samples were cultured in yeast nitrogen base (YNB)-D-xylose or YNB-xylan media. Candida tropicalis, Asterotremella humicola, Candida boidinii and Debaryomyces hansenii were the most frequently isolated yeasts. Among D-xylose-fermenting yeasts, six strains of Spathaspora passalidarum, two of Scheffersomyces stipitis, and representatives of five new species were identified. The new species included Candida amazonensis of the Scheffersomyces clade and Spathaspora sp. 1, Spathaspora sp. 2, Spathaspora sp. 3, and Candida sp. 1 of the Spathaspora clade. In fermentation assays using D-xylose (50 g/L) culture medium, S. passalidarum strains showed the highest ethanol yields (0.31 g/g to 0.37 g/g) and productivities (0.62 g/L.h to 0.75 g/L.h). Candida amazonensis exhibited a virtually complete D-xylose consumption and the highest xylitol yields (0.55 g/g to 0.59 g/g), with concentrations up to 25.2 g/L. The new Spathaspora species produced ethanol and/or xylitol in different concentrations as the main fermentation products. In sugarcane bagasse hemicellulosic fermentation assays, S. stipitis UFMG-XMD-15.2 generated the highest ethanol yield (0.34 g/g) and productivity (0.2 g/L.h), while the new species Spathaspora sp. 1 UFMG-XMD-16.2 and Spathaspora sp. 2 UFMG-XMD-23.2 were very good xylitol producers. Conclusions/Significance: This study demonstrates the promise of using new D-xylose-fermenting yeast strains from the Brazilian Amazonian Forest for ethanol or xylitol production from sugarcane bagasse hemicellulosic hydrolysates.

Spiroplasma in Drosophila melanogaster Populations: Prevalence, Male-Killing, Molecular Identification, and No Association with Wolbachia

Spiroplasma endosymbionts are maternally transmitted bacteria that may kill infected sons resulting in the production of female-biased broods. The prevalence of male killers varies considerably both between and within species. Here, we evaluate the spatial and temporal status of male-killing and non-male-killing Spiroplasma infection in three Brazilian populations of Drosophila melanogaster, nearly a decade after the first occurrence report for this species. The incidence of the male-killing Spiroplasma ranged from close to 0 to 17.7 % (so far the highest estimate for a Drosophila species) with a suggestion of temporal decline in a population. We also found non-male-killing Spiroplasma coexisting in one population at lower prevalence (3-5 %), and we did not detect it in the other two. This may be taken as a suggestion of a spreading advantage conferred by the male-killing strategy. Sequencing two loci, we identified the phylogenetic position of Spiroplasma strains from the three localities, showing that all strains group closely in the poulsonii clade. Due to intensive sampling effort, we were able to test the association between Spiroplasma infections and another widespread endosymbiont, Wolbachia, whose prevalence ranged from 81.8 to 100 %. The prevalence of Wolbachia did not differ between Spiroplasma-infected and uninfected strains in our largest sample nor were the prevalences of the two endosymbionts associated across localities.

The Diversity of Polyketide Synthase Genes from Sugarcane-Derived Fungi

The chemical ecology and biotechnological potential of metabolites from endophytic and rhizosphere fungi are receiving much attention. A collection of 17 sugarcane-derived fungi were identified and assessed by PCR for the presence of polyketide synthase (PKS) genes. The fungi were all various genera of ascomycetes, the genomes of which encoded 36 putative PKS sequences, 26 shared sequence homology with beta-ketoacyl synthase domains, while 10 sequences showed homology to known fungal C-methyltransferase domains. A neighbour-joining phylogenetic analysis of the translated sequences could group the domains into previously established chemistry-based clades that represented non-reducing, partially reducing and highly reducing fungal PKSs. We observed that, in many cases, the membership of each clade also reflected the taxonomy of the fungal isolates. The functional assignment of the domains was further confirmed by in silico secondary and tertiary protein structure predictions. This genome mining study reveals, for the first time, the genetic potential of specific taxonomic groups of sugarcane-derived fungi to produce specific types of polyketides. Future work will focus on isolating these compounds with a view to understanding their chemical ecology and likely biotechnological potential.