Caracterização filogenética de percevejos terrestres das famílias Coreidae e Pentatomidae (Heteroptera: Pentatomomorpha) por meio de marcadores moleculares

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Phylogeographic analysis in populations of Protonectarina sylveirae (Saussure, 1854) (Hymenoptera, Vespidae, Polistinae)

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

Evolution of river dolphins

The world's river dolphins (Inia, Pontoporia, Lipotes and Platanista) are among the least known and most endangered of all cetaceans. The four extant genera inhabit geographically disjunct river systems and exhibit highly modified morphologies, leading many cetologists to regard river dolphins as an unnatural group. Numerous arrangements have been proposed for their phylogenetic relationships to one another and to other odontocete cetaceans. These alternative views strongly affect the biogeographical and evolu- tionary implications raised by the important, although limited, fossil record of river dolphins. We present a hypothesis of river dolphin relationships based on phylogenetic analysis of three mitochondrial genes for 29 cetacean species, concluding that the four genera represent three separate, ancient branches in odontocete evolution. Our molecular phylogeny corresponds well with the first fossil appearances of the primary lineages of modern odontocetes. Integrating relevant events in Tertiary palaeoceanography, we develop a scenario for river dolphin evolution during the globally high sea levels of the Middle Miocene. We suggest that ancestors of the four extant river dolphin lineages colonized the shallow epicontinental seas that inundated the Amazon, Parana, Yangtze and Indo-Gangetic river basins, subsequently remaining in these extensive waterways during their transition to freshwater with the Late Neogene trend of sea-level lowering.

Phylogeographic analysis in populations of Protonectarina sylveirae (Saussure, 1854) (Hymenoptera, Vespidae, Polistinae)

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

LOW GENETIC DIVERSITY IN Wolbachia-INFECTED Culex quinquefasciatus (DIPTERA: CULICIDAE) FROM BRAZIL AND ARGENTINA

Culex quinquefasciatus is a vector of human pathogens, including filarial nematodes and several viruses. Although its epidemiological relevance is known to vary across geographical regions, an understanding of its population genetic structure is still incipient. In light of this, we evaluated the genetic diversity of Cx. quinquefasciatus and Cx. pipiens x Cx. quinquefasciatus hybrids collected from nine localities in Brazil and one site in Argentina. We used mitochondrial genes cox1 and nd4, along with the coxA and wsp genes of the maternally-inherited Wolbachia endosymbiont. The nd4 fragment was invariant between samples, whilst cox1 exhibited four haplotypes that separated two types of Cx. quinquefasciatus, one clustered in southern Brazil. Low sequence diversity was generally observed, being discussed. Both Brazilian and Argentinian mosquitoes were infected with a single Wolbachia strain. As reported in previous studies with these populations, cox1 and nd4 diversity is not congruent with the population structure revealed by nuclear markers or alar morphology. Future Cx. quinquefasciatus research should, if possible, evaluate mtDNA diversity in light of other markers.

A relict new species of Oreobates (Anura, Strabomantidae) from the Seasonally Dry Tropical Forests of Minas Gerais, Brazil, and its implication to the biogeography of the genus and that of South American Dry Forests

A new species of Oreobates is described from Cavernas do Peruacu National Park, Januaria, Minas Gerais state, in the Atlantic Dry Forests of Brazil. The new species is distinguished from all other Oreobates by having the following combination of characters: large tympanum, discs broadly enlarged and truncate on Fingers III and IV, smooth dorsal skin, nuptial pads absent, snout subacuminate, and a very short pulsatile (2-3 pulses) single-noted advertisement call with dominant frequency of about 3150 Hz, and no harmonic structure. Molecular phylogenetic analyses using partial sequences of the mitochondrial genes cytochrome b (cyt b) and 16S using multiple outgroups recovered the new species within Oreobates and sister to O. heterodactylus. The latter species inhabits the Dry Forests of Mato Grosso (Cerrado) and Bolivia (Chiquitano forests), and is strictly associated to these habitats, which suggests a preterit connection between Chiquitano and Atlantic Dry Forests. The discovery of a new Oreobates in the Atlantic Dry Forest is of great importance for the conservation of these dry forests, as it is known only from this type of habitat.

Cytogenetic and molecular analyses in troglobitic and epigean species of Pimelodella (Siluriformes: Heptapteridae) from Brazil

Samples from seven different locations of the genus Pimelodella were genetically examined, two caves (exclusively subterranean, upper Tocantins River and Sao Francisco River) and five epigean (from upper Parana River basin). Cytogenetic analyses revealed the same diploid number (2n=46) for all species besides similarities in both number and location of nucleolar organizer regions and C bands. FISH with 5S rDNA probes and CMA(3) staining indicated significant differences among the studied species. Application of PCR-RFLP in ATPase 6 and 8 mitochondrial genes allowed building a minimum evolution phenogram identifying the close evolutionary relationship among groups. Both chromosomal and molecular data were useful to infer the relationships among studied Pimelodella species.

Polyphyly of the hawk genera Leucopternis and Buteogallus (Aves, Accipitridae): multiple habitat shifts during the Neotropical buteonine diversification

Abstract Background The family Accipitridae (hawks, eagles and Old World vultures) represents a large radiation of predatory birds with an almost global distribution, although most species of this family occur in the Neotropics. Despite great morphological and ecological diversity, the evolutionary relationships in the family have been poorly explored at all taxonomic levels. Using sequences from four mitochondrial genes (12S, ATP8, ATP6, and ND6), we reconstructed the phylogeny of the Neotropical forest hawk genus Leucopternis and most of the allied genera of Neotropical buteonines. Our goals were to infer the evolutionary relationships among species of Leucopternis, estimate their relationships to other buteonine genera, evaluate the phylogenetic significance of the white and black plumage patterns common to most Leucopternis species, and assess general patterns of diversification of the group with respect to species' affiliations with Neotropical regions and habitats. Results Our molecular phylogeny for the genus Leucopternis and its allies disagrees sharply with traditional taxonomic arrangements for the group, and we present new hypotheses of relationships for a number of species. The mtDNA phylogenetic trees derived from analysis of the combined data posit a polyphyletic relationship among species of Leucopternis, Buteogallus and Buteo. Three highly supported clades containing Leucopternis species were recovered in our phylogenetic reconstructions. The first clade consisted of the sister pairs L. lacernulatus and Buteogallus meridionalis, and Buteogallus urubitinga and Harpyhaliaetus coronatus, in addition to L. schistaceus and L. plumbeus. The second clade included the sister pair Leucopternis albicollis and L. occidentalis as well as L. polionotus. The third lineage comprised the sister pair L. melanops and L. kuhli, in addition to L. semiplumbeus and Buteo buteo. According to our results, the white and black plumage patterns have evolved at least twice in the group. Furthermore, species found to the east and west of the Andes (cis-Andean and trans-Andean, respectively) are not reciprocally monophyletic, nor are forest and non-forest species. Conclusion The polyphyly of Leucopternis, Buteogallus and Buteo establishes a lack of concordance of current Accipitridae taxonomy with the mtDNA phylogeny for the group, and points to the need for further phylogenetic analysis at all taxonomic levels in the family as also suggested by other recent analyses. Habitat shifts, as well as cis- and trans-Andean disjunctions, took place more than once during buteonine diversification in the Neotropical region. Overemphasis of the black and white plumage patterns has led to questionable conclusions regarding the relationships of Leucopternis species, and suggests more generally that plumage characters should be used with considerable caution in the taxonomic evaluation of the Accipitridae.

Low genetic diversity in Wolbachia-Infected Culex quinquefasciatus (Diptera: Culicidae) from Brazil and Argentina

Culex quinquefasciatus is a vector of human pathogens, including filarial nematodes and several viruses. Although its epidemiological relevance is known to vary across geographical regions, an understanding of its population genetic structure is still incipient. In light of this, we evaluated the genetic diversity of Cx. quinquefasciatus and Cx. pipiens x Cx. quinquefasciatus hybrids collected from nine localities in Brazil and one site in Argentina. We used mitochondrial genes cox1 and nd4, along with the coxA and wsp genes of the maternally-inherited Wolbachia endosymbiont. The nd4 fragment was invariant between samples, whilst cox1 exhibited four haplotypes that separated two types of Cx. quinquefasciatus, one clustered in southern Brazil. Low sequence diversity was generally observed, being discussed. Both Brazilian and Argentinian mosquitoes were infected with a single Wolbachia strain. As reported in previous studies with these populations, cox1 and nd4 diversity is not congruent with the population structure revealed by nuclear markers or alar morphology. Future Cx. quinquefasciatus research should, if possible, evaluate mtDNA diversity in light of other markers.

Phylogeny of Western Palaearctic long-eared bats (Mammalia, Chiroptera, Plecotus) - a molecular perspective

Die Phylogenie der Westpaläarktischen Langohren (Mammalia, Chiroptera, Plecotus) – eine molekulare Analyse Die Langohren stellen eine Fledermausgattung dar, die fast alle westpaläarktischen Habitate bist zum Polarkreis hin besiedeln und in vielerlei Hinsicht rätselhaft sind. In der Vergangenheit wurden zahlreiche Formen und Varietäten beschrieben. Trotzdem galt für lange Zeit, dass nur zwei Arten in Europa anerkannt wurden. Weitere Arten waren aus Nordafrika, den Kanaren und Asien bekannt, aber auch deren Artstatus wurde vielfach in Frage gestellt. In der vorliegenden Dissertation habe ich mittels molekularer Daten,der partiellen Sequenzierung mitochondrialer Gene (16S rRNA und ND1), sowie der mitochondrialen Kontrollregion, eine molekular Analyse der phylogenetischen Verwandtschaftsverhältnisse innerhalb und zwischen den Linien der westpaläarktischen Langohren durchgeführt. Die besten Substitutionsmodelle wurden berechnet und phylogenetische Bäume mit Hilfe vier verschiedener Methoden konstruiert: dem neighbor joining Verfahren (NJ), dem maximum likelihood Verfahren (ML), dem maximum parsimony Verfahren (MP) und dem Bayesian Verfahren. Sechs Linien der Langohren sind genetisch auf einem Artniveau differenziert: Plecotus auritus, P. austriacus, P. balensis, P. christii, P. sardus, und P. macrobullaris. Im Falle der Arten P. teneriffae, P. kolombatovici und P. begognae ist die alleinige Interpretation der genetischen Daten einzelner mitochondrialer Gene für eine Festlegung des taxonomischen Ranges nicht ausreichend. Ich beschreibe in dieser Dissertation drei neue Taxa: Plecotus sardus, P. kolombatovici gaisleri (=Plecotus teneriffae gaisleri, Benda et al. 2004) and P. macrobullaris alpinus [=Plecotus alpinus, Kiefer & Veith 2002). Morphologische Kennzeichen, insbesondere für die Erkennung im Feld, werden hier dargestellt. Drei der sieben Arten sind polytypisch: P. auritus (eine west- und ein osteuropäische Linie, eine sardische Linie und eine aktuell entdeckte kaukasische Linie, Plecotus kolombatovici (P. k. kolombatovici, P. k. gaisleri und P. k. ssp.) und P. macrobullaris (P. m. macrobullaris und P. m. alpinus). Die Verbreitungsgebiete der meisten Arten werden in dieser Arbeit erstmals ausschließlich anhand genetisch zugeordneter Tiere dargestellt.Die Untersuchung der ökologischen Einnischung der nun anerkannten Formen, insbesondere in Gebieten sympatrischer Verbreitung, bietet ein spannendes und lohnendes Feld für zukünftige Forschungen. Nicht zuletzt muss sich die Entdeckung eines beachtlichen Anteils kryptischer Diversität innerhalb der westpaläarktischen Langohren auch bei der Entwicklung spezieller Artenschutzkonzepte widerspiegeln.

Phylogenomische Analysen bei Metazoen - Zur Stellung der Xenoturbellida und Syndermata

Im Rahmen der vorliegenden Dissertation wurde die phylogenetischen Stellungen der Xenoturbellida (Deuterostomia) und der Syndermata (Protostomia) mit phylogenomischen Techniken untersucht. Auf methodischer Ebene konnte gezeigt werden, dass ribosomale Proteine aufgrund ihres mittleren bis hohen Konservierungsgrades, ihrer Häufigkeit in kleineren EST-Projekten, damit verbunden ihrer Häufigkeit in Datenbanken und ihres phylogenetischen Informationsgehalts nützliche Werkzeuge für phylogenetische Fragestellungen sind. Es konnte durch phylogenetische Rekonstruktionen und Hypothesentests auf Basis eines 11.912 Aminosäuren langen Datensatzes gezeigt werden, dass die Xenoturbellida innerhalb der Deuterostomia eine Schwestergruppenbeziehung zu den Ambulacraria eingehen. Diese Arbeit zeigt im Vergleich aller bisher durchgeführten Arbeiten die beste statistische Unterstützung für diese Topologie. Weiterhin konnte untermauert werden, dass die Urochordata vermutlich anstelle der Cephalochordata die Schwestergruppe der Vertebrata sind. Der Vergleich der publizierten Xenoturbella EST-Datensätze mit dem eigenen Datensatz ließ den Rückschluß zu, dass ESTs offenbar klar weniger anfällig gegen Kontaminationen mit Erbmaterial (DNA+RNA) anderer Spezies sind als PCR-Amplifikate genomischer oder mitochondrialer Gene. Allerdings bestimmt anscheinend der physiologische Zustand der Tiere die Repräsentation von Transkriptklassen wie Stressproteine und mitochondriale Transkripte. Die bakteriellen Transkripte in einem der EST-Datensätze stammen vermutlich von Chlamydien, die möglicherweise symbiontisch in Xenoturbella bocki leben. Im Bereich der Protostomia wurden drei EST-Projekte für Vertreter der Syndermata durchgeführt. Basierend auf drei verschiedenen Proteinalignment-Datensätzen von ca. 11.000 Aminosäuren Länge konnte gezeigt werden, dass die Syndermata innerhalb der Spiralia einzugruppieren sind und dass sie mit den Gnathostomulida das monophyletische Supertaxon Gnathifera bilden. Die genaue phylogenetische Position der Syndermata innerhalb der Spiralia konnte hingegen noch nicht eindeutig geklärt werden, ebenso wie kein kongruenter Beweis für die Existenz des Supertaxons Platyzoa gefunden werden konnte. Im Rahmen der Untersuchung der internen Phylogenie der Syndermata konnten drei der fünf konkurrierenden Hypothesen aufgrund der Paraphylie der Eurotatoria ausgeschlossen werden. Da keine Daten der Seisonidea in den Analysen implementiert waren, bleibt die Frage der internen Phylogenie der Syndermata letztlich offen. Klar ist jedoch, dass die Eurotatoria nicht wie bislang angenommen monophyletisch sind, da die räderorgantragenden Bdelloidea keinesfalls den morphologisch diesbezüglich ähnlichen Monogononta ähnlich sind, sondern den räderorganlosen Acanthocephala näher stehen. Die Abbildung der molekularen Phylogenie auf die morphologischen Verhältnisse zeigt, dass das Räderorgan (partiell oder komplett) offenbar kurz nach der Aufspaltung der Syndermata in Monogononta und Acanthocephala + Bdelloidea in der Acanthocephala + Bdelloidea-Linie reduziert wurde. Die Entstehung des einziehbaren hinteren Körperteils (Rostrum bei Bdelloidea bzw. Proboscis bei Acanthocephala) in der Acanthocephala + Bdelloidea-Linie könnte das Schlüsselereignis zur Entstehung des Endoparasitismus der Acanthocephala gewesen sein.

RNA editing in kinetoplastids

RNA editing in kinetoplastid protozoa is a post-transcriptional process of uridine insertion or deletion in mitochondrial mRNAs. The process involves two RNA species, the pre-edited mRNA and in most cases a trans-acting guide RNA (gRNA). Sequences within gRNAs define the position and extend of mRNA editing. Both mRNAs and gRNAs are encoded by mitochondrial genes in the kinetoplast DNA (kDNA), which consists of thousands of small circular DNA molecules, called minicircles, encoding thousands of gRNAs, catenated together and with a few mRNA encoding larger circles, the maxicircles, to form a huge DNA network. Editing has been shown to result in translatable mRNAs of bona fide mitochondrial genes as well as novel alternatively edited transcripts that are involved in the maintenance of the kDNA itself. RNA editing occurs within large protein-RNA complexes, editosomes, containing gRNA, preedited and partially edited mRNAs and also structural and catalytically active proteins. Editosomes are diverse in both RNA and protein composition and undergoe structural remodeling during the maturation. The compositional and structural diversity of editosomes further underscores the complexity of the RNA editing process.

Molecular Phylogenetics of Mole Crabs (Hippidae : Emerita)

Mole crabs of the genus Emerita (Family Hippidae) inhabit many of the temperate and tropical sandy beaches of the world. The nine described species of this genus are rarely sympatric, and most are endemic to broad biogeographic regions. The phylogenetic relationships among the species have not yet been investigated. Based on presumed morphological synapomorphics, it has been suggested that the species inhabiting the New World constitute a monophyletic group, as do the species inhabiting the Old World, The relationships within the New World species were previously studied using sequence data from Cytochrome Oxidase I and 16S rRNA mitochondrial genes; the results strongly suggested that one of the species, Emerita analoga, was very divergent from the other taxa examined. This observation prompted uncertainty about monophyly of the New World species. The goal of the present study was to elucidate the relationships among the species within the genus Emerita. Partial sequences for the mitochondrial COI and 16S rRNA genes for all nine species of the genus (and several outgroups) were examined. Phylogenetic analyses suggest that E. analoga is closer to the Old World taxa than to the other New World species; thus the New World Emerita species do not constitute a monophyletic group.

Phylogenetic relationships of the hoatzin, an enigmatic South American bird.

The hoatzin (Opisthocomus hoazin) lives in the humid lowlands of northern and central South America, often in riparian habitats. It is a slender bird approximately 65 cm in length, brownish with lighter streaks and buffy tips to the long tail feathers. The small head has a ragged, bristly crest of reddish-brown feathers, and the bare skin of the face is bright blue. It resembles a chachalaca (Ortalis, Cracidae) in size and shape, but its plumage and markings are similar to those of the smaller guira cuckoo (Guira guira). The hoatzin (pronounced Watson) has been a taxonomic puzzle since it was described in 1776. It usually has been viewed as related to the gallinaceous birds, but alliances to other groups have been suggested, including the cuckoos. We present DNA sequence evidence from the 12S and 16S rRNA mitochondrial genes, and from the nuclear gene that codes for the eye lens protein, alpha A-crystallin. The results indicate that the hoatzin is most closely related to the typical cuckoos and that the divergence occurred at or near the base of the cuculiform phylogenetic tree.

Variabilidade populacional em manguezais: análises moleculares e morfológicas em caranguejos Brachyura (Crustacea: Decapoda)

A análise da estruturação populacional de espécies codistribuídas permite a comparação de padrões de estruturação, fornecendo informações acerca dos fatores que influenciam a diferenciação em espécies pertencentes ao mesmo ecossistema. Este projeto teve como objetivo principal analisar a variabilidade intraespecífica em caranguejos habitantes de manguezais, codistribuídos ao longo do Oceano Atlântico Ocidental, por meio de ferramentas moleculares e morfológicas, visando testar a hipótese de elevada estruturação populacional em manguezais. Para este fim, cinco espécies foram utilizadas como modelo (Aratus pisonii, Goniopsis cruentata, Sesarma rectum, Uca thayeri e Ucides cordatus) e avaliadas por meio de marcadores mitocondriais COI e 16S e nuclear H3 e análises morfológicas comparativas e de morfometria. Os dados moleculares revelaram dois padrões, indicando elevada estruturação populacional para as espécies A. pisonii e U. thayeri e ausência de estruturação para G. cruentata, S. rectum e U. cordatus. Os dados morfológicos, no entanto, não acompanham esses padrões, já que não foram encontradas diferenças morfológicas ou morfométricas associadas aos grupos evidenciados pelas análises moleculares. A ausência de fluxo gênico entre regiões para algumas espécies deve-se, muito provavelmente, à existência de fatores que não se limitam ao isolamento por distância, mas também devido a diferenças na duração do estágio larval e a diferenças bruscas em alguns fatores abióticos, como a salinidade, por exemplo, que, associados às diferentes características do desenvolvimento larval de cada espécie, culminam na existência de estruturas populacionais diferentes. Além disso, os padrões de diferenciação genética observados concordam com os cenários biogeográficos propostos para o Atlântico Ocidental, no qual mudanças e flutuações geológicas, climáticas e oceanográficas, resultantes do fechamento do Istmo do Panamá e de ciclos glaciais na América do Norte, promoveram divergência genética.