Sistemática molecular de Atta laevigata (Smith 1858) e Acromyrmex balzani (Emery 1890)

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

Molecular phylogenies, chromosomes and dispersion in Brazilian akodontines (Rodentia, Sigmodontinae)

Uma nova filogenia molecular para roedores akodontinos do Brasil é proposta. A árvore filogenética foi enriquecida com a área de ocorrência e com informações sobre o cariótipo das amostras. Baseado nisto, e com a metodologia descrita, foram propostas hipóteses sobre as características do cariótipo e sobre a área de ocorrência dos ancestrais de cada clado. Assim, foi possível discutir hipóteses sobre evolução cromossômica do grupo, e sobre eventos de dispersão a partir da área de diferenciação original dos akodontinos nos Andes. A evolução cromossômica começou com números diplóides altos (2n=52) e mostrou uma tendência a redução (até 2n=14 em clados mais recentes). Ramos independentes da árvore mostraram redução do 2n e num caso aumentou o numero diplóide. Foram propostos pelo menos quatro eventos de dispersão dos Andes até o Brasil Sul-Oriental. Os resultados indicam a direção de novos estudos em cariologia comparada.

Molecular phylogeny, morphology and their implications for the taxonomy of Eriocaulaceae

The pantropical family Eriocaulaceae includes ten genera and c. 1,400 species, with diversity concentrated in the New World. The last complete revision of the family was published more than 100 years ago, and until recently the generic and infrageneric relationships were poorly resolved. However, a multi-disciplinary approach over the last 30 years, using morphological and anatomical characters, has been supplemented with additional data from palynology, chemistry, embryology, population genetics, cytology and, more recently, molecular phylogenetic studies. This led to a reassessment of phylogenetic relationships within the family. In this paper we present new data for the ITS and trnL-F regions, analysed separately and in combination, using maximum parsimony and Bayesian inference. The data confirm previous results, and show that many characters traditionally used for differentiating and circumscribing the genera within the family are homoplasious. A new generic key with characters from various sources and reflecting the current taxonomic changes is presented.

Anatomía comparada de la madera de Cupressaceae y su correspondencia con los estudios de filogenia

La familia Cupressaceae incluye un total de 133 especies agrupadas en 30 géneros, 17 de los cuales son monospecíficos. Esta familia se encuentra representada en todos los continentes salvo en la Antártida. Sus especies se distribuyen en distintas regiones climáticas, y en altitudes que varían desde el nivel del mar hasta los 5.000 m. La falta de descripción anatómica de muchos de los géneros y especies de Cupressaceae es notable, así como la contradicción que aparece entre distintas investigaciones sobre las características anatómicas de la madera descritas para cada especie. Este estudio describe la anatomía de la madera de Cupressaceae y analiza las características que podrían representar sinapomorfías de los clados delimitados en los estudios filogenéticos. Siguiendo los métodos tradicionales de preparación y descripción de la madera a nivel microscópico, se ha estudiado la madera de 113 especies de los 30 géneros de Cupressaceae. Para ello se han empleado muestras de madera de origen trazable, procedentes de colecciones de madera de distintas instituciones internacionales. Se ha empleado una robusta filogenia molecular para la reconstrucción de los caracteres ancestrales. La anatomía de la madera de los 30 géneros de Cupressaceae, pone de manifiesto la gran homogeneidad de la familia, caracterizada por la presencia de traqueidas axiales sin engrosamientos helicoidales, parénquima radial con paredes horizontales lisas, punteaduras del campo de cruce de tipo cupresoide y la carencia de canales resiníferos fisiológicos. Además, todos presentan parénquima axial (salvo Neocallitropsis, Thuja y Xanthocyparis), punteaduras radiales areoladas con toro definido (salvo Thuja y Thujopsis), siendo habitual la presencia de punteaduras areoladas en las paredes tangenciales de la madera tardía, y verrugosidades en la cara interna de las traqueidas (salvo Ca. macleayana, Libocedrus, Papuacedrus y Neocallitropsis). Los radios leñosos son homogéneos y están compuestos de parénquima radial (con la presencia de traqueidas radiales en algunas especies de Cupressus, Sequoia, Thujopsis y X. nootkatensis) con paredes finales lisas o lisas y noduladas (exclusivamente noduladas en Cal. macrolepis, C. bakeri y en la mayoría de especies de Juniperus), y el rango de altura de los radios leñosos se encuentra entre 5 y 15 células. Se consideran posibles sinapomorfismos de Cupressaceae la presencia de verrugosidades en la cara interna de las traqueidas, la presencia de traqueidas axiales sin engrosamientos helicoidales, la presencia de parénquima axial, la presencia de radios leñosos homogéneos (compuestos únicamente de parénquima radial), la tipología de las paredes horizontales del parénquima radial, las punteaduras del campo de cruce de tipo cupresoide y la ausencia de canales resiníferos fisiológicos, pero lo que realmente diferencia a este grupo de coníferas es la simultaneidad de todos estos caracteres en sus maderas. Como sinapomorfías específicas por clados se proponen: la ausencia de toro definido y muescas en el borde de las punteaduras en Thuja-Thujopsis, la existencia de extensiones de toro en Diselma-Fitzroya-Widdringtonia; la presencia de engrosamientos callitroides en Callitris-Actinostrobus; la presencia de espacios intercelulares y las muescas en el borde de las punteaduras en el clado formado por el género Juniperus y las especies de Cupressus en la región oriental; la presencia de paredes finales del parénquima radial tanto lisas como noduladas en los clados formados por el género Xanthocyparis y las especies de Cupressus en la región occidental y en Fitzroya-Diselma; y por último, la presencia de punteaduras del campo de cruce de tipo taxodioide en los clados taxodioid y sequoioid. ABSTRACT The Cupressaceae family comprises 133 species grouped into 30 genera, 17 of which are monotypic. The family is represented in all continents except Antarctica. Its species are distributed in various climate zones and at altitudes from sea level to 5,000 m. There is a considerable lack of anatomical descriptions for many genera and species of Cupressaceae and much contradiction between studies about the wood anatomical features described for each species. This study describes the wood anatomy of Cupressaceae and analyses the features that could represent synapomorphies of the clades recovered in phylogenetic studies. Following the traditional methods of preparation and description of wood at microscopic level, a study was made of the wood of 113 species of the 30 Cupressaceae genera. The study samples had traceable origins and came from wood collections held at various international institutions. A robust molecular phylogeny was used for ancestral state reconstruction. The wood anatomy of the 30 genera of the Cupressaceae shows the high homogeneity of the family, which is characterised by the presence of axial tracheids without helical thickenings, smooth horizontal walls of ray parenchyma cells, cupressoid cross-field pits, and the absence of physiological resin canals. In addition, they all have axial parenchyma (except Neocallitropsis, Thuja and Xanthocyparis), a warty layer on the inner wall of the tracheids (except Ca. macleayana, Libocedrus, Papuacedrus and Neocallitropsis) and tracheid pitting in radial walls with a well defined torus (except Thuja and Thujopsis); tracheid pitting in the tangential walls of the latewood is common. Rays are homogeneous and are composed of ray parenchyma (with the presence of ray tracheids in some species of Cupressus, Sequoia, Thujopsis and X. nootkatensis), with smooth end walls or both smooth and nodular end walls (exclusively nodular in Cal. macrolepis, C. bakeri and most Juniperus species), and ray height range is 5 to 15 cells. Possible synapomorphies of Cupressaceae are the presence of a warty layer on the inner layer of the tracheids, axial tracheids without helical thickenings, the presence of axial parenchyma, homogeneous rays (composed exclusively of ray parenchyma), the typology of the horizontal walls of ray parenchyma cells, cupressoid cross-field pits and the absence of physiological resin canals, but what truly differentiates this group of softwoods is the co-occurrence of all these features in their wood. The following are proposed as clade-specific synapomorphies: absence of a well-defined torus and presence of pits with notched borders in Thuja-Thujopsis, torus extensions in Diselma-Fitzroya-Widdringtonia; callitroid thickenings in Callitris-Actinostrobus; intercellular spaces and pits with notched borders in the clade formed by the genus Juniperus and the species of Cupressus in the eastern region; smooth and nodular ray parenchyma end walls in the clades formed by the genus Xanthocyparis and the species of Cupressus in the western region and in Fitzroya-Diselma, and taxodioid cross-field pits in the taxodioid and sequoioid clades.

Identification of novel molecular determinants of tissue mineralization in fish

Tese de doutoramento, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2015

Diversidade genética em um grupo de anuros em miniatura endêmico da Mata Atlântica brasileira (Euparkerella Griffiths 1959).

A Mata Atlântica (MA) está entre as regiões com maior biodiversidade e mais ameaçadas do planeta. Esforços em diversas áreas do conhecimento têm sido feitos para que se tenha uma estimativa mais refinada da diversidade existente e sua organização ao longo do bioma. O crescente número de estudos que buscam reconstituir a história da diversificação da MA apontam para um cenário espacial e temporal complexo, havendo ainda uma lacuna no conhecimento dos processos em pequena escala. Vertebrados em miniatura têm se mostrado uma boa ferramenta para estudos de processos evolutivos em pequena escala. Assim, o gênero Euparkerella, endêmico de uma pequena região da MA dos Estados do Rio de Janeiro (RJ) e Espírito Santo (ES), foi escolhido como modelo para este estudo. No primeiro capítulo buscou-se descrever a diversidade existente dentro do gênero a partir de uma filogenia molecular. Para isso, utilizaram-se métodos bayesianos para gerar genealogias de genes e de espécies a partir de um fragmento de gene mitocondrial e quatro fragmentos de genes nucleares. Os resultados obtidos apontaram para uma grande diversidade críptica no gênero. Foram identificadas seis unidades evolutivas significativamente divergentes para o RJ: duas em Euparkerella cochranae, três em Euparkerella brasiliensis, e Euparkerella sp.. A espécie mais basal recuperada foi Euparkerella robusta, do ES, e estimou-se o início da diversificação do gênero para o final do Mioceno. O segundo capítulo descreve onze marcadores de microssatélites desenvolvidos para Euparkerella brasiliensis através do método de pirosequenciamento de nova geração 454. No terceiro capítulo estudou-se apenas uma unidade evolutiva, Euparkerella brasiliensis da área dos Três Picos/ RJ. A partir de marcadores de evolução rápida (microssatélites) e lenta (sequências de DNA) buscou-se compreender a estrutura e a dinâmica populacional desta unidade evolutiva em uma área bastante pequena (aprox. 20 km) sob influência de um gradiente ambiental altitudinal (40 m 1000 m). Foram identificadas, a partir dos microssatélites, duas subpopulações geneticamente distintas nas bordas do gradiente. O fluxo gênico se deu predominantemente das bordas para a zona de contato, onde foi observado o maior efetivo populacional. Tais resultados indicam que pequenas variações ambientais podem atuar no isolamento populacional em Euparkerella e corroboram o padrão de formas microendêmicas identificadas na filogenia. Futuros estudos devem ser feitos no sentido de buscar caracterizar morfologicamente as unidades evolutivas aqui identificadas; preencher as lacunas amostrais, especialmente no ES; e descrever os processos que atuam em pequena escala nas zonas de contato entre as unidades evolutivas e fatores limitantes a distribuição das mesmas.

Systematic and evolutionary studies of the terrestrial gastropods Helicoidea and Pyramidula

178 p.

Evolution, gene regulation and functional analysis of BMP2 in fish

Bone morphogenetic proteins (BMPs) are multifunctional growth factors belonging to the transforming growth factor β (TGFβ) superfamily with a central role in bone formation and mineralization. BMP2, a founding member of this family, has demonstrated remarkable osteogenic properties and is clinically used to promote bone repair and fracture healing. Lack of basic data on factors regulating BMP2 expression and activity have hampered a better understanding of its role in bone formation and bone-related diseases. The objective of this work was to collect new functional data and determine spatiotemporal expression patterns in a fish system aiming towards a better understanding of BMP2 function and regulation. Transcriptional and post-transcriptional regulation of gilthead seabream BMP2 gene was inferred from luciferase reporter systems. Several bone- and cartilage-related transcription factors (e.g. RUNX3, MEF2c, SOX9 and ETS1) were found to regulate BMP2 transcription, while microRNA 20a was shown to affect stability of the BMP2 transcript and thus the mineralogenic capacity of fish bone-derived host cells. The regulation of BMP2 activity through an interaction with the matrix Gla protein (MGP) was investigated in vitro using BMP responsive elements (BRE) coupled to luciferase reporter gene. Although we demonstrated the functionality of the experimental system in a fish cell line and the activation of BMP signaling pathway by seabream BMP2, no conclusive evidence could be collected on a possible interaction beween MGP and BMP2. The evolutionary relationship among the members of BMP2/4/16 subfamily was inferred from taxonomic and phylogenetic analyses. BMP16 diverged prior to BMP2 and BMP4 and should be the result of an ancient genome duplication that occurred early in vertebrate evolution. Structural and functional data suggested that all three proteins are effectors of the BMP signaling pathway, but expression data revealed different spatiotemporal patterns in teleost fish suggesting distinct mechanisms of regulation. In this work, through the collection of novel data, we provide additional insight into the regulation, the structure and the phylogenetic relationship of BMP2 and its closely related family members.

Espécies crípticas em Paracoccidioides brasiliensis: diferenciação antigênica e de resposta a fatores estressantes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Relacionamento filogenético de espécies pertencentes às famílias Coreidae e Pentatomidae (Heteroptera) a partir dos genes mitocondriais COI, 16S e nuclear 18S

Pós-graduação em Genética - IBILCE

The prion protein and New World primate phylogeny

The PrPC prion protein contains 250 amino acids with some variation among species and is expressed in several cell types. PrPC is converted to PrPSc by a post-translational process in which it acquires amino acid sequences of three-dimensional conformation of -sheets. Variations in the prion protein gene were observed among 16 genera of New World primates (Platyrrhini), and resulted in amino acid substitutions when compared with the human sequence. Seven substitutions not yet described in the literature were found: W  R at position 31 in Cebuella, T  A at position 95 in Cacajao and Chiropotes, N S at position 100 in Brachyteles, L  Q at position 130 in Leontopithecus (in the sequence responsible for generating the -sheet 1), D  E at position 144 in Lagothrix (in the sequence responsible for the -helix 1), D G at position 147 in Saguinus (also located in the -helix 1 region), and M  I at position 232 in Alouatta. The phylogenetic trees generated by parsimony, neighbor-joining and Bayesian analyses strongly support the monophyletic status of the platyrrhines, but did not resolve relationships among families. However, the results do corroborate previous findings, which indicate that the three platyrrhine families radiated rapidly from an ancient split.

Low levels of genetic diversity depicted from mitochondrial DNA sequences in a heavily exploited marine fish (Cynoscion acoupa, Sciaenidae) from the Northern coast of Brazil

The acoupa weakfish (Cynoscion acoupa - Sciaenidae) is a marine species of croaker with estuarine-dependent behavior, found in the western Atlantic from Panama to Argentina. It is one of the most exploited food fish on the northern coast of Brazil. In this study, DNA sequences were determined from the entire control region (D-loop) of the mitochondrial genome of 297 individuals collected during seven different months between December 2003 and August 2005 on the northern coast of Brazil (Amapá and Pará). Genetic variability expressed by haplotype (h = 0,892) and nucleotide (p = 0,003) diversities were low compared to other heavily exploited marine fish species from the western Atlantic and eastern Asia. AMOVA depicted a lack of genetic structuring among the samples from different years, indicating the presence of a single stock of C. acoupa within the sample area. The possible reasons for the low levels of genetic diversity are discussed. These results demonstrate a need for the monitoring of C. acoupa harvesting and the preservation of the estuaries within its geographic range, considering that this large fish depends on estuarine ecosystems during part of its life cycle.

Tocantins river as an effective barrier to gene flow in Saguinus niger populations

The Saguinus represent the basal genus of the Callitrichinae subfamily. Traditionally this genus is divided into three groups: Hairy, Mottled and Bare-face, however, molecular data failed to validate these groups as monophyletic units, as well as raised some subspecies to the species status. This is the case of the former subspecies Saguinus midas midas and S. midas niger, which are now considered as different species. In the present study, we sequenced a portion of the D-loop mtDNA region in populations from the East bank of the Xingu and from both banks of the Tocantins river, in order to test the effectiveness of large rivers as barriers to the gene flow in Saguinus. According to our results, the populations from the East and West banks of the Tocantins river are more divergent than true species like S. mystax and S. imperator. The Tocantins river may be acting as a barrier to gene flow, and consequently these very divergent populations may represent distinct taxonomic entities (species?).

Phylogenetic analysis of 16S mitochondrial DNA data in sloths and anteaters

We sequenced part of the 16S rRNA mitochondrial gene in 17 extant taxa of Pilosa (sloths and anteaters) and used these sequences along with GenBank sequences of both extant and extinct sloths to perform phylogenetic analysis based on parsimony, maximum-likelihood and Bayesian methods. By increasing the taxa density for anteaters and sloths we were able to clarify some points of the Pilosa phylogenetic tree. Our mitochondrial 16S results show Bradypodidae as a monophyletic and robustly supported clade in all the analysis. However, the Pleistocene fossil Mylodon darwinii does not group significantly to either Bradypodidae or Megalonychidae which indicates that trichotomy best represents the relationship between the families Mylodontidae, Bradypodidae and Megalonychidae. Divergence times also allowed us to discuss the taxonomic status of Cyclopes and the three species of three-toed sloths, Bradypus tridactylus, Bradypus variegatus and Bradypus torquatus. In the Bradypodidae the split between Bradypus torquatus and the proto-Bradypus tridactylus / B. variegatus was estimated as about 7.7 million years ago (MYA), while in the Myrmecophagidae the first offshoot was Cyclopes at about 31.8 MYA followed by the split between Myrmecophaga and Tamandua at 12.9 MYA. We estimate the split between sloths and anteaters to have occurred at about 37 MYA.

A new species of Mesocoelium (Digenea: Mesocoeliidae) found in Rhinella marina (Amphibia: Bufonidae) from Brazilian Amazonia

ABSTRACT: Mesocoelium lanfrediae sp. nov. (Digenea: Mesocoeliidae) inhabits the small intestine of Rhinella marina (Amphibia: Bufonidae) and is described here, with illustrations provided by light, scanning electron microscopy and molecular approachs. M. lanfrediae sp. nov. presents the typical characteristics of the genus, but is morphometrically and morphologically different from the species described previously. The main diagnostic characteristics of M. lanfrediae sp. nov. are (i) seven pairs of regularly-distributed spherical papillae on the oral sucker, (ii) ventral sucker outlined by four pairs of papillae distributed in a uniform pattern and interspersed with numerous spines, which are larger at the posterior margin and (iii) small, rounded tegumentary papillae around the opening of the oral sucker, which are morphologically different from those of the oral sucker itself, some of which are randomly disposed in the ventrolateral tegumentary region of the anterior third of the body. Addionally, based on SSU rDNA, a phylogenetic analysis including Brachycoeliidae and Mesocoeliidae taxa available on GenBank established the close relationship between M. lanfrediae sp. nov. and Mesocoelium sp.