The evolutionary relationships in Xenopus genus are intensively studied for its interspecific variability and high conservation in evolution. These characteristics possess an opportunity for comparative studying of polyploidization phenomenom on interchromosomal level and an occasion to identify the genome-forming mechanisms with cytogenetic methods. XME chromosomes (X. mellotropicalis, 2n=40) were identified via p-/q- arm length ratio in a comparison with morphometric analysis of X. epitropicalis (2n=40) chromosomes. Whole chromosome painting probes were prepared from X. tropicalis (2n=20) microdissected chromosomes and they were applied to XME metaphase spreads via optimalized Zoo-FISH. 10 chromosomal quartets were detected and one balanced non-reciprocal translocation between chromosomes XME 2 and XME 9 which must have occured in a diploid ancestor. Thus, we disprove the theory of Silurana subgenus origin via only one polyploidizatin event. Powered by TCPDF (www.tcpdf.org)

Made available in DSpace on 2014-06-11T19:32:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-18Bitstream added on 2014-06-13T20:03:23Z : No. of bitstreams: 1 cabraldemello_d_dr_botib.pdf: 1654082 bytes, checksum: 7e6b12e5a0a939210a7949c8a83f729c (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O mapeamento cromossômico de seqüências repetitivas de DNA tem se mostrado uma eficiente ferramenta nos estudos comparativos e evolutivos em diversos organismos. Estudos cromossômicos com besouros da subfamília Scarabaeinae têm revelado ampla variabilidade, entretanto a análise da organização cromossômica de DNAs repetitivos neste grupo é escassa e direcionada unicamente ao mapeamento do DNA ribossomal (DNAr) 18S. O presente trabalho teve como objetivo caracterizar cromossomicamente DNAs repetitivos em espécies de Scarabaeinae, utilizando bandeamentos cromossômicos e mapeamento físico cromossômico de seqüências repetitivas, incluindo famílias multigênicas de RNAr 18S, RNAr 5S e histona H3 e a fração de DNA C0t-1. Ampla variabilidade foi observada relacionada ao número/localização dos sítios de DNAr 18S, aparentemente associada a diversificação da heterocromatina. Por outro lado, os genes de RNAr 5S e histona H3, mostraram-se amplamente conservados e co-localizados em um par cromossômico, com aparente intercalação. Análises em representantes de Dichotomius revelaram conservação dos blocos de heterocromatina, entretanto com aparente compartimentalização dos mesmos. O uso da fração DNA C0t-1 confirmou o enriquecimento em DNAs repetitivos da heterocromatina, que se apresentou diversificada entre as espécies, utilizando como referência D. geminatus. Por outro lado, regiões terminais dos cromossomos apresentaram-se amplamente conservadas entre as seis espécies. Além disso, a análise da fração de DNAs repetitivos em D. geminatus indicou origem intraespecífica do cromossomo B desta espécie que possivelmente pode estar sofrendo homogeneização com seqüências encontradas no complemento A. Os resultados indicam distintos padrões de diversificação para o DNA repetitivo nos representantes de Scarabaeinae, sugerindo extensiva reorganizaçãomicrogenômica ao longo / The chromosomal mapping of repeated DNAs has been used as an efficient tool in comparative and evolutionary studies in some organism. The chromosomal studies in beetles belonging to the subfamily Scarabaeinae have revealed wide variability, although the analysis of chromosomal organization of repeated DNAs in this group is scarce and directed solely for 18S rDNA mapping. The present study aimed in chromosomal characterization of repeated DNAs in Scarabaeinae species using chromosomal banding and physical chromosome mapping of repeated sequences, including the multigene families for 18S and 5S rRNAs and H3 histone genes and the C0t-1 DNA fraction. Wide variability was observed concerning the number and location of 18S rDNA sites, apparently associated to the heterochromatin diversification. On the other hand, the 5S rRNA and H3 histone genes were widely conserved and co-located in one chromosomal pair, showing apparently interspersion. Analysis in Dichotomius representatives revealed conservation for heterochromatic blocks, although an apparent compartmentalization was observed. The use of C0t-1 DNA fraction confirmed the heterochromatin repeated DNAs enrichment, which is diversified among the species, using as reference D. geminatus. On the other hand, the terminal regions of the chromosomes were highly conserved among the six species. Moreover, the analysis of repeated DNA fraction from D. geminatus indicated intraspecific origin of a B chromosome in this species that possibly could be suffering homogenization with A complement sequences. The results indicate distinct diversification patterns for repeated DNAs in Scarabaeinae representatives, suggesting extensive microgenomic reorganization along the cladogenesis of the group

International malaria control initiatives such as the Roll Back Malaria Initiative (RBM) and the Medicines for Malaria Venture (MMV) mobilize resources and spur research aimed at vector control as well as the treatment and eventual eradication of the disease. These efforts have managed to reduce incidence of malaria by an estimated 37% worldwide since 2000. However, despite the promising success of control efforts such as these, the World Health Organization reports a staggering 438,000 deaths from malaria in 2015. The continuing high death toll of malaria as well as emerging insecticide and antimalarial drug resistance suggests that while encouraging, success in reducing malaria incidence may be tenuous. Current vector control strategies are often complicated by ecological and behavioral heterogeneity of vector mosquito populations. As an additional obstruction, mosquito genomes are highly plastic as evidenced by the wealth or chromosomal inversions that have occurred in this genus. Chromosomal inversions have been correlated with differences in adaptation to aridity, insecticide resistance, and differences in resting behavior. However, a good understanding of the molecular mechanisms for inversion generation is still lacking. One possible contributor to inversion formation in Anopheles mosquitoes includes repetitive DNA such as transposable elements (TEs), tandem repeats (TRs) and inverted repeats (IRs). This dissertation provides physical maps for two important malaria vectors, An. stephensi and An. albimanus (Ch.2 and Ch. 3) and then applies those maps to the identification of inversion breakpoints in malaria mosquitoes. Repeat content of each chromosomal arm and the molecular characterization of lineage specific breakpoints is also investigated (Ch. 2 and Ch.4). Our study reveals differences in patterns of chromosomal evolution of Anopheles mosquitoes vs. Drosophila. First, mosquito chromosomes tend to shuffle as intact elements via whole arm translocations and do not under fissions or fusions as seen in fruitflies. Second, the mosquito sex chromosome is changing at a much higher rate relative to the autosomes in malaria mosquitoes than in fruit flies. Third, our molecular characterization of inversion breakpoints indicates that TEs and TRs may participate in inversion genesis in an arm specific manner. / Ph. D. / Malaria is a complex and devastating disease vectored by the bite of a female Anopheles mosquito. This disease claimed an estimated 438,000 lives in 2015. The mobilization of funding and resources as part of global malaria eradication initiatives have reduced the global incidence of malaria by 37% in the last 15 years. Deaths from malaria are also 60% lower vs. the year 2000. These promising gains are threatened by the ability of Anopheles mosquitoes to adapt in the face of malaria control efforts. Anopheles mosquito chromosomes are known to be highly plastic, as evidenced by numerous chromosomal inversions. Recent years have seen increases in insecticide resistance, and behavioral change in mosquito populations that allow them to avoid insecticides and remain prolific vectors of disease. This ability of mosquito vectors to adapt threatens to unravel recent progress towards a malaria free world. The projects presented in this dissertation explore mechanisms of chromosomal evolution, specifically the potential role of repetitive DNA in the generation of chromosomal inversions. The exploration of chromosomal inversions was facilitated by the creation of physical maps for Anopheles species. Prominent malaria vectors An. stephensi andAn. albimanus were physically mapped in Chapter 2 and Chapter 3 respectively. In chapter 1 and chapter 3 physical maps are utilized for the identification of chromosomal inversion breakpoints using 2 species (Ch. 2) and many species (Ch. 4). Repeat content was quantified along each chromosomal arm (Ch 2,4) and in inversion breakpoint regions (Ch 3). This dissertation presents physical maps for two important malaria species that have been applied to the study of chromosomal evolution and will also serve as community tools for further study of malaria mosquitoes. Our work on chromosomal evolution has revealed the Anopheles chromosomes tend to undergo translocations as intact elements and do not under fissions and fusions as seen in fruitflies. We also find that the malaria mosquito sex chromosome changes much more rapidly relative to the autosomes than in fruitflies. Additionally, repetitive DNA including transposable elements (TEs) and tandem repeats (TRs) may be encouraging chromosomal inversions but with differing roles on different chromosomal arms.

Resumo: O mapeamento cromossômico de seqüências repetitivas de DNA tem se mostrado uma eficiente ferramenta nos estudos comparativos e evolutivos em diversos organismos. Estudos cromossômicos com besouros da subfamília Scarabaeinae têm revelado ampla variabilidade, entretanto a análise da organização cromossômica de DNAs repetitivos neste grupo é escassa e direcionada unicamente ao mapeamento do DNA ribossomal (DNAr) 18S. O presente trabalho teve como objetivo caracterizar cromossomicamente DNAs repetitivos em espécies de Scarabaeinae, utilizando bandeamentos cromossômicos e mapeamento físico cromossômico de seqüências repetitivas, incluindo famílias multigênicas de RNAr 18S, RNAr 5S e histona H3 e a fração de DNA C0t-1. Ampla variabilidade foi observada relacionada ao número/localização dos sítios de DNAr 18S, aparentemente associada a diversificação da heterocromatina. Por outro lado, os genes de RNAr 5S e histona H3, mostraram-se amplamente conservados e co-localizados em um par cromossômico, com aparente intercalação. Análises em representantes de Dichotomius revelaram conservação dos blocos de heterocromatina, entretanto com aparente compartimentalização dos mesmos. O uso da fração DNA C0t-1 confirmou o enriquecimento em DNAs repetitivos da heterocromatina, que se apresentou diversificada entre as espécies, utilizando como referência D. geminatus. Por outro lado, regiões terminais dos cromossomos apresentaram-se amplamente conservadas entre as seis espécies. Além disso, a análise da fração de DNAs repetitivos em D. geminatus indicou origem intraespecífica do cromossomo B desta espécie que possivelmente pode estar sofrendo homogeneização com seqüências encontradas no complemento A. Os resultados indicam distintos padrões de diversificação para o DNA repetitivo nos representantes de Scarabaeinae, sugerindo extensiva reorganizaçãomicrogenômica ao longo / Abstract: The chromosomal mapping of repeated DNAs has been used as an efficient tool in comparative and evolutionary studies in some organism. The chromosomal studies in beetles belonging to the subfamily Scarabaeinae have revealed wide variability, although the analysis of chromosomal organization of repeated DNAs in this group is scarce and directed solely for 18S rDNA mapping. The present study aimed in chromosomal characterization of repeated DNAs in Scarabaeinae species using chromosomal banding and physical chromosome mapping of repeated sequences, including the multigene families for 18S and 5S rRNAs and H3 histone genes and the C0t-1 DNA fraction. Wide variability was observed concerning the number and location of 18S rDNA sites, apparently associated to the heterochromatin diversification. On the other hand, the 5S rRNA and H3 histone genes were widely conserved and co-located in one chromosomal pair, showing apparently interspersion. Analysis in Dichotomius representatives revealed conservation for heterochromatic blocks, although an apparent compartmentalization was observed. The use of C0t-1 DNA fraction confirmed the heterochromatin repeated DNAs enrichment, which is diversified among the species, using as reference D. geminatus. On the other hand, the terminal regions of the chromosomes were highly conserved among the six species. Moreover, the analysis of repeated DNA fraction from D. geminatus indicated intraspecific origin of a B chromosome in this species that possibly could be suffering homogenization with A complement sequences. The results indicate distinct diversification patterns for repeated DNAs in Scarabaeinae representatives, suggesting extensive microgenomic reorganization along the cladogenesis of the group / Orientador: Cesar Martins / Coorientador: Rita de Cássia de Moura / Banca: Marcelo dos Santos Guerra / Banca: Orlando Moreira Filho / Banca: Sanae Kasahara / Banca: Maria Cristina de Almeida / Doutor

Made available in DSpace on 2015-03-26T13:42:17Z (GMT). No. of bitstreams: 1 texto completo.pdf: 4676037 bytes, checksum: 95dde20b76c33bfd09f530eb8a65e6e5 (MD5) Previous issue date: 2010-07-19 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The tribe Attini consists of a monophyletic group that includes about 230 species in 14 genera. Different biological substrate are utilized by these ants for cultivation of the symbiotic fungus on which they feed and are necessarily dependent. This tribe includes the leaf-cutting ants (Atta Fabricius and Acromyrmex Mayr) that are known as important agricultural pests. The usage of cytogenetics in the family Formicidae has made significant contributions in the study of more than 750 ant species. Only 10% of Attini’s species have some sort of cytogenetic data available. This work aimed at cytogenetically studying six Acromyrmex species and seven species of the tribe Attini divided into four genera: Apterostigma Mayr, Mycocepurus Forel, Sericomyrmex Mayr and Trachymyrmex Forel. Some of the species were also submitted to banding techniques. The karyotypes observed were: Apterostigma madidiense Weber (n=23, 7m + 10sm + 5st + 1a), Apterostigma steigeri Santschi (2n=22, 20m + 2sm), Mycocepurus goeldii (2n=8, 4m + 4sm), Sericomyrmex sp. (2n=50, 44m + 6 sm; n=25, 22m + 3 sm), Trachymyrmex fuscus Emery (2n=18, 16m + 2sm; n=9, 8m + 1sm), Trachymyrmex relictus Borgmeier (2n=20m, n=10m) and Trachymyrmex sp. (2n=22, 18m + 4sm). A size polymorphism on a short arm of a submetacentric pair from Trachymyrmex fuscus was also pointed out. The possible causes of chromosomal segment duplication were discussed. All studied species of the genus Acromyrmex presented 2n=38. Each species showed a different karyotype in relation to morphology (A. balzani: 12m + 10sm + 14st + 2a; A. coronatus: 12m + 12sm + 12st + 2a; A. disciger: 10m + 12sm + 14st + 2a; A. niger: 12m + 18sm + 6st + 2a; A. rugosus: 16m + 12sm + 8st + 2a; A. echinatior: 8m + 6sm + 14st + 10a), in the distribution of heterocromatin (C banding) and GC rich regions (CMA3). For three species (A. coronatus, A. disciger, A. niger) the first subtelocentric pair was the bearer of ribosomal genes. Acromyrmex species presented stability in the number of chromosome with differences in the karyotype. The results suggest that many chromosomal rearrangements such as translocations, growth of heterochromatin and inversion may have occurred during their differentiation, contributing to the karyotype variability found. / A tribo Attini é um grupo monofilético que inclui mais de 230 espécies agrupadas em 14 gêneros. Nesta tribo estão incluídas as formigas cortadeiras (Atta e Acromyrmex), consideradas importantes pragas agrícolas. A utilização da citogenética na família Formicidae tem mostrado importante contribuição nas mais de 750 espécies de formigas estudadas. Os dados citogenéticos são reportados para 10% das espécies da tribo Attini. O presente trabalho teve como objetivo o estudo citogenético de seis espécies do gênero Acromyrmex e de sete espécies de Attini distribuídas em quatro gêneros: Apterostigma, Mycocepurus, Sericomyrmex e Trachymyrmex. Parte das espécies foi submetida a técnicas de bandamentos cromossômicos. Os cariótipos observados foram: Apterostigma madidiense Weber (n=23, 7m + 10sm + 5st + 1a), A. steigeri Santschi (2n=22, 20m + 2sm), Mycocepurus goeldii (Forel) (2n=8, 4m + 4sm), Sericomyrmex sp. (2n=50, 44m + 6sm; n=25, 22m + 3sm), Trachymyrmex fuscus Emery (2n=18, 16m + 2sm; n=9, 8m + 1sm), T. relictus Borgmeier (2n=20m, n=10m) e Trachymyrmex sp. (2n=22, 18m + 4sm). Trachymyrmex fuscus apresentou polimorfismo de tamanho no braço curto do par submetacêntrico. As possíveis causas da duplicação do segmento cromossômico são discutidas. As seis espécies do gênero Acromyrmex apresentaram 2n=38 cromossomos. Cada espécie mostrou um cariótipo diferente em relação à morfologia (A. balzani: 12m + 10sm + 14st + 2a; A. coronatus: 12m + 12sm + 12st + 2a; A. disciger: 10m + 12sm + 14st + 2a; A. niger: 12m + 18sm + 6st + 2a; A. rugosus: 16m + 12sm + 8st + 2a; A. echinatior: 8m + 6sm + 14st + 10a) e variações na distribuição de heterocromatina (banda C) e regiões ricas em GC (CMA3). Em três espécies (A. coronatus, A. disciger, A. niger) o par subtelocêntrico de maior tamanho foi o portador de genes ribossomais. As espécies do gênero Acromyrmex apresentaram estabilidade no número cromossômico com diferenças no cariótipo. Isto sugere que rearranjos do tipo translocações, crescimento de heterocromatina e inversão devem ter ocorrido durante a evolução das espécies deste gênero, levando a modificações nos cariótipos das diferentes espécies.

In this work, the integrative taxonomy approach was performed to understand species limits and patterns of diversification in two genera of orizomyine rodents (Cerradomys and Oligoryzomys). Therefore, molecular markers with distinct evolutionary rates were used with different approaches (phylogeny, coalescent-based species delimitation, DNA barcoding, phylogeography, molecular dating). Classic and molecular cytogenetic analyzes were performed, contributing to cytotaxonomy and revealing chromosomal evolution. This work is divided into four chapters, including a brief introduction (Chapter 1). In Chapter 2, the integrative taxonomy approach was used to study the genus Cerradomys, based on cytogenetic and molecular data. The results revealed that cytogenetics is important in the recognition of all described species (cytotaxonomy). Phylogenetic reconstruction showed that internal relationships are well supported, with the exception of C. subflavus and C. goytaca, which are not reciprocally monophyletic. Following the integrative taxonomy, in which species limits are based on the congruence of methods, this work recognizes and reiterates the eight Cerradomys species described so far. We suggest a taxonomic revision in C. langguthi and C. subflavus, since both may represent species-complex or in process of speciation. Times of divergence show that Cerradomys is a recent genus, with speciation events occurred mainly in the Pleistocene. In Chapter 3, classic and molecular cytogenetics (Fluorescence in situ hybridization - FISH with telomeric and Oligoryzomys moojeni probes) were used to study chromosomal evolution in Cerradomys, based on the molecular phylogeny obtained in Chapter 2. Chromosome painting revealed extensive chromosome reshuffling in Cerradomys. Species with the highest diploid numbers showed exclusively telomeric signals whereas interstitial telomeric signals (ITS) were observed in the species with the lowest diploid numbers. Comparisons of chromosome painting with molecular phylogeny data corroborate the hypothesis that ITS, in this case, are remnants of telomeres. Nevertheless, other chromosomal rearrangements were detected with absence of ITS, indicating that these sequences may have been lost in the process of chromosomal breakages, evidencing that there was both retention and loss of ITS along the karyotypic evolution of the genus. In addition, complex rearrangements were detected between the karyotypes of C. goytaca and C. subflavus, reiterating that these two species are distinct, since hybrids probably would not be viable due to meiotic problems. In Chapter 4, aiming to recover the evolutionary history and species limits of Oligoryzomys, molecular phylogeny studies were integrated into cytogenetic data. The genus was monophyletic, but the internal relations had low support. The compilation of phylogenetic, chromosomal data and geographic distribution (interdisciplinarity) was important to understand species boundaries. Four lineages could not be related to any name and may be new species (Oligoryzomys A-D). Oligoryzomys flavescens was recovered paraphyletic in respect to O. fornesi. Oligoryzomys stramineus, O. microtis and O. nigripes were recovered in two well-structured clades each. In the case of the last two species, the subclades are probably related to exclusive karyotypes. In O. microtis, one subclade is composed of samples from the western Amazon region and the other with samples distributed in southern Amazon region, transition with Cerrado (2n=64, FN=64). In O. nigripes, one of the clades is composed of specimens from northeastern Brazil (2n=62, FN=78) and the other from central-south-southeast Brazil, Argentina, Paraguay and Uruguay (2n=62, FN=80-82). Phylogeographic results corroborate phylogenetic and cytogenetic data, revealing two distinctive phylogroups, consistent with incipient species. Chromosome data corroborate previous work and could be associated to the following names: O. mattogrossae, O. moojeni, O. chacoensis, O. stramineus, O. nigripes and O. flavescens, although the last two species should be reassessed. In addition, an undescribed karyotype is being reported for Oligoryzomys aff. utiaritensis (2n=70, FN=72), as well as new records in Brazil for four species. We suggest a taxonomic revision in O. microtis, O. flavescens and O. nigripes, as these species probably represent incipient or species-complex. In addition, samples related to Oligoryzomys aff. delicatus, Oligoryzomys aff. chacoensis, Oligoryzomys aff. rupestris and Oligoryzomys aff. Utiaritensis should be evaluated morphologically to confirm their identities. The results of this work corroborate the importance of interdisciplinary studies, since the rates of evolution differ according to each character / Neste trabalho, utilizou-se a abordagem de taxonomia integrativa para compreender os limites das espécies e padrão de diversificação em dois gêneros de roedores orizominos (Cerradomys e Oligoryzomys). Para tanto, marcadores moleculares com taxas evolutivas distintas foram utilizados em diferentes abordagens (filogenia, delimitação de espécies baseada em coalescência, DNA barcoding, filogeografia, datação). Análises de citogenética clássica e molecular foram realizadas, contribuindo como um marcador citotaxonômico e revelando padrões de evolução cromossômica. Os dados moleculares e citogenéticos, combinados à dados de distribuição geográfica, tornaram esse trabalho interdisciplinar. Esta tese está dividida em quatro capítulos, incluindo uma breve introdução (Capítulo 1). No capítulo 2, a abordagem de taxonomia integrativa foi utilizada para estudar o gênero Cerradomys, a partir dos dados citogenéticos e moleculares. Os resultados revelaram que a citogenética é importante no reconhecimento de todas as espécies descritas (citotaxonomia). A reconstrução filogenética mostrou que as relações internas são bem suportadas, com exceção de C. subflavus e C. goytaca, que não são reciprocamente monofiléticos. De acordo com a taxonomia integrativa, em que a delimitação de espécies é baseada na congruência entre a maioria dos dados, esse trabalho reconhece e reitera as oito espécies de Cerradomys descritas até o momento. Sugerimos uma revisão taxonômica em C. langguthi e C. subflavus, uma vez que ambas podem representar complexos de espécies ou casos de especiação em curso. Os tempos de divergência mostram que Cerradomys é um gênero recente, cujos eventos de especiação ocorreram preponderantemente no Pleistoceno. No capítulo 3, estudos de citogenética clássica e molecular (hibridação in situ fluorescente - FISH com sondas teloméricas e cromossomo-específicas de Oligoryzomys moojeni) foram realizados para compreender a evolução cromossômica de Cerradomys, com base na filogenia obtida no capítulo anterior. A pintura cromossômica mostrou que um grande número de rearranjos ocorreu ao longo da evolução cariotípica de Cerradomys. As espécies com os maiores números diplóides mostraram sinais exclusivamente teloméricos enquanto que sinais teloméricos intersticiais (ITS) foram observados nas espécies com menores números diplóides. Comparações dos dados de pintura cromossômica com os dados de filogenia molecular corroboram a hipótese de que as ITS, neste caso, são remanescentes de telômeros. No entanto, outros rearranjos cromossômicos foram detectados com ausência de ITS, de modo que essas sequências podem ter sido perdidas no processo das quebras cromossômicas, evidenciando que houve tanto retenção quanto perda das ITS ao longo da evolução cariotípica do gênero. Além disso, rearranjos complexos foram detectados entre os cariótipos de C. goytaca e C. subflavus, reiterando que essas duas espécies são distintas, uma vez que provavelmente os híbridos não seriam viáveis devido a problemas meióticos. No capítulo 4, com o objetivo de recuperar a história evolutiva e os limites das espécies de Oligoryzomys, estudos de filogenia molecular foram integrados a dados citogenéticos. O gênero mostrou-se monofilético, mas as relações internas tiveram baixo suporte. A compilação dos dados filogenéticos, cromossômicos e de distribuição geográfica (interdisciplinaridade) foram importantes para compreender os limites das espécies. Quatro linhagens não puderam ser relacionadas a nenhum nome, sendo prováveis espécies novas (Oligoryzomys A-D). Oligoryzomys flavescens foi recuperado parafilético em relação à O. fornesi. Oligoryzomys stramineus, O. microtis e O. nigripes foram recuperados em dois clados bem estruturados cada. No caso das duas últimas espécies, os subclados provavelmente estão relacionados à cariótipos exclusivos. Em O. microtis, um dos clados é composto por exemplares do oeste da região amazônica e o outro, por exemplares distribuído ao sul da região amazônica, transição com Cerrado (2n=64, NF=64). Em O. nigripes, um dos clados é composto por exemplares do nordeste do Brasil (2n=62, NF=78) e o outro por exemplares da região centro-sul-sudeste do Brasil, Argentina, Paraguai e Uruguai (2n=62, NF=80-82). Os dados filogeográficos suportam os dados filogenéticos e cromossômicos, revelando dois filogrupos em O. nigripes, sugerindo que essas populações estejam em processo de especiação. Os dados cromossômicos corroboram as informações da literatura e puderam ser associados aos seguintes nomes: O. mattogrossae, O. moojeni, O. chacoensis, O. stramineus, O. flavescens e O. nigripes, embora as duas últimas devam ser reavaliadas. Adicionalmente, um novo cariótipo está sendo reportado para Oligoryzomys aff. utiaritensis (2n=70, NF=72), assim como novos dados de distribuição no Brasil para quatro espécies. Sugerimos uma revisão taxonômica em O. microtis, O. flavescens e O. nigripes, pois estas espécies provavelmente representam complexos de espécies ou estão em processo de especiação. Além disso, os exemplares relacionados à Oligoryzomys aff. delicatus, Oligoryzomys aff. chacoensis, Oligoryzomys aff. rupestris e Oligoryzomys aff. utiaritensis devem ser avaliados morfologicamente para confirmar suas identidades. Os resultados desse trabalho corroboram a importância dos estudos interdisciplinares, uma vez que as taxas de evolução para cada caráter são heterogêneas

<p>The common shrew, <i>Sorex araneus</i>, has one of the most variable karyotypes among mammals, displaying numerous chromosomes races throughout its distribution, which can be categorized into different karyotypic groups. The objective of this thesis was to examine the postglacial population history of Fennoscandian common shrews using autosomal microsatellites, mitochondrial DNA (mtDNA) and a Y chromosome specific microsatellite (L8Y).</p><p>Autosomal microsatellites and mtDNA revealed weak genetic structure over a hybrid zone between the karyotypically divergent Northern and Western karyotypic groups. However, the genetic structure displayed by the Y chromosome microsatellite was orders of magnitude higher. Hence, considerable chromosomal differences between the groups do not prevent female gene flow, while male gene flow is reduced (cf. Haldane's rule). Further, the results suggest that the Haldane effect may be caused by the chromosomal differences between the karyotypic groups.</p><p>No mtDNA differentiation was observed either between chromosome races or between the Northern and Western karyotypic groups in Fennoscandia. The combined pattern of karyotypic and mtDNA variation of Fennoscandian common shrews, suggest bi-directional postglacial recolonisation from a single refugium in Europe. The variation of the Y-linked microsatellite supported this conclusion. In contrast, significant mtDNA structure, discordant with the karyotypic variation, revealed that common shrews in southern Finland belong to a different lineage than remaining Fennoscandian regions, implying postglacial recolonisation from a different source.</p><p>MtDNA variation of the chromosome races in Sweden supports the hypothesis that three races of the Western karyotypic group have been formed through whole arm reciprocal translocations (WARTs), as suggested by their mutual karyotypic variation. The variation of the molecular markers supports the theory of rapid karyotypic evolution in the common shrew.</p>

The common shrew, Sorex araneus, has one of the most variable karyotypes among mammals, displaying numerous chromosomes races throughout its distribution, which can be categorized into different karyotypic groups. The objective of this thesis was to examine the postglacial population history of Fennoscandian common shrews using autosomal microsatellites, mitochondrial DNA (mtDNA) and a Y chromosome specific microsatellite (L8Y). Autosomal microsatellites and mtDNA revealed weak genetic structure over a hybrid zone between the karyotypically divergent Northern and Western karyotypic groups. However, the genetic structure displayed by the Y chromosome microsatellite was orders of magnitude higher. Hence, considerable chromosomal differences between the groups do not prevent female gene flow, while male gene flow is reduced (cf. Haldane's rule). Further, the results suggest that the Haldane effect may be caused by the chromosomal differences between the karyotypic groups. No mtDNA differentiation was observed either between chromosome races or between the Northern and Western karyotypic groups in Fennoscandia. The combined pattern of karyotypic and mtDNA variation of Fennoscandian common shrews, suggest bi-directional postglacial recolonisation from a single refugium in Europe. The variation of the Y-linked microsatellite supported this conclusion. In contrast, significant mtDNA structure, discordant with the karyotypic variation, revealed that common shrews in southern Finland belong to a different lineage than remaining Fennoscandian regions, implying postglacial recolonisation from a different source. MtDNA variation of the chromosome races in Sweden supports the hypothesis that three races of the Western karyotypic group have been formed through whole arm reciprocal translocations (WARTs), as suggested by their mutual karyotypic variation. The variation of the molecular markers supports the theory of rapid karyotypic evolution in the common shrew.

L'étude comparative de l'architecture des génomes mammaliens a révélé l'association des séquences répétées et des réarrangements. Cette thèse porte sur la dynamique et le rôle dans les remaniements de deux types de séquences répétées: les clusters ribosomiques et les satellites. Ces séquences sont analysées par une approche cytogénomique (FISH, CO-FISH) dans le genre Mus connu pour sa diversité chromosomique, et pour lequel les phylogénies moléculaires et chromosomiques sont disponibles.1) La distribution chromosomique des clusters ribosomiques, établie chez 19 espèces, a permis de reconstruire les états ancestraux des clusters. Cette analyse montre que les clusters (24%) sont associés à des points de cassures, mais présentent également une grande labilité en l'absence de réarrangements. De plus, une forte association entre les clusters et les centromères est mise en évidence. 2) Le sous-genre Mus se caractérise par un caryotype très conservé excepté chez une sous-espèce de la souris domestique (M. musculus domesticus), qui est connue pour son extraordinaire radiation chromosomique impliquant les séquences satellites du centromère. Afin de rechercher les spécificités génomiques responsables de ce patron d'évolution contrasté, la dynamique évolutive des séquences satellites a été analysée chez 11 taxons. Révélant des différences qualitatives entre taxons, cette étude a permis de proposer un scénario évolutif de ces séquences. Toutefois, aucune des caractéristiques étudiées (composition, orientation) n'est propre à M. m. domesticus, et ne permet de rendre compte de sa plasticité chromosomique. De même, chez cette dernière, aucun lien entre la quantité de séquences satellites et la fréquence d'implication des chromosomes dans les réarrangements n'est mis en évidence.Cette étude confirme que les séquences répétées participent à l'évolution chromosomique, mais ne constituent pas à elles seules l'élément clef de cette dernière. / Comparative analyses of the architecture of mammalian genomes have highlighted the association between repetitive sequences and rearrangements. This thesis focuses on the evolutionary dynamics of two repeat sequences (ribosomal clusters and satellites) and explores their role in chromosomal change. These sequences are analyzed by a cytogenomic approach (FISH, CO-FISH) in the genus Mus that is known for its chromosomal diversity and for which molecular and chromosomal phylogenies are available.1) The chromosomal distribution of ribosomal clusters, established in 19 species, allowed us to reconstruct the ancestral states of clusters. This analysis demonstrated that 24% of clusters were associated with breakpoints, whereas others showed high lability in the absence of rearrangements. Moreover, a strong association between clusters and centromeres was retrieved.2) The subgenus Mus is characterized by a highly conserved karyotype except for one subspecies of the house mouse (M. musculus domesticus), that displays an extraordinary chromosomal radiation involving centromeric satellite sequences. To determine the genomic traits related to this difference in rate, the evolutionary dynamics of satellite sequences was analyzed in 11 taxa. From the qualitative differences evidenced between taxa, an evolutionary scenario of these sequences is proposed. None of the studied features (composition, orientation) of these sequences was found to be specific to M. m. domesticus, and could explain its chromosomal plasticity. Similarly, in the latter, no relationship between satellite sequence quantity and the rearrangement frequency of chromosomes was found.This study confirms that although repeated sequences are involved in chromosomal evolution, they aren't in themselves the key element of the latter.

Made available in DSpace on 2014-12-17T14:33:04Z (GMT). No. of bitstreams: 1 PabloAM.pdf: 4013213 bytes, checksum: 97a52b7c01ae798889ede43cc9641282 (MD5) Previous issue date: 2010-02-26 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Given the great diversity of fishes, the Order Tetraodontiformes stands to show genetic and morphological characteristics enough singular. The fishes of this order have a compact DNA which favors molecular studies, as well as comparisons with more basal species. Model of genome evolution, there are still many gaps in knowledge about their chromosomal patterns and how evolutionary rearrangements influence the marked variation in DNA content of this order. In view of this, we present cytogenetic analyzes of the species Acanthostracion quadricornis (Ostraciidae), A. polygonius (Ostraciidae) Melichthys niger (Balistidae) Cantherhines macrocerus (Monacanthidae) and C. pullus (Monacanthidae), Lagocephalus laevigatus, Colomesus psittacus and Canthigaster figueiredoi (Tetraodontidae), to contribute with cytogenetic data for this group. The analysis was performed by C-banding, Ag-RONs, coloring with base-specific fluorochromes DAPI-CMA3, restriction enzymes AluI, EcoRI, TaqI, PstI and HinfI and in situ hybridization with probes for ribosomal DNA 18S and 5S. The heterochromatic ultrastructure of A. quadricornis and A. polygonius revealed a outstanding heterochromatin content, which may indicate that the accumulation or loss of extensive heterochromatin content could be responsible for large variations in genomic content displayed in different Tetraodontiformes families. The species Cantherhines macrocerus, C. pullus (Monacanthidae) and Melichthys niger (Balistidae) shows a huge karyotypic similarity both numerically and structural. L. laevigatus showed similar cytogenetic features (2n = 44 and single RONs) to the species of the genus Takifugu, which reinforces the idea of their phylogenetic relationships. C. psittacus presented the highest diploid number described for the family (2n = 56) and large amount of HC, features that related with its sister family Diodontidae. Cytogenetic analysis in C. figueiredoi revealed heterochromatic polymorphisms, RONs multiple and Bs chromosomes. These events are rare in marine fishes, and are possibly associated with the strong restructuring and genomic reduction that this family has been suffered. These features, plus the morphological and molecular data suggests that these species share the same ancestral branch, with a possible monophyletic origin. In this study, new contributions to the knowledge of evolutionary patterns facing by Tetraodontiformes are provided and discussed under cytotaxonomyc, genomic and evolutionary perspectives. / Frente ? grande diversidade de peixes, a Ordem Tetraodontiformes se destaca por exibir caracter?sticas gen?ticas e morfol?gicas bastantes singulares. Os peixes desta Ordem apresentam um DNA compacto o que favorece estudos moleculares, assim como compara??es com esp?cies mais basais. Modelo de evolu??o gen?mica, ainda existem v?rias lacunas de conhecimento sobre seus padr?es cromoss?micos e como os rearranjos evolutivos influenciaram na marcante varia??o no conte?do de DNA desta Ordem. Diante disto o presente estudo apresenta an?lises citogen?ticas das esp?cies, Acanthostracion quadricornis (Ostraciidae), A. polygonius (Ostraciidae), Melichthys niger (Balistidae), Cantherhines macrocerus (Monacanthidae), C. pullus (Monacanthidae), Lagocephalus laevigatus, Colomesus psittacus e Canthigaster figueiredoi (Tetraodontidae) visando contribuir com mais dados citogen?ticos para o grupo. As an?lises foram realizadas atrav?s do bandamento C, Ag-RONs, colora??o com fluorocromos base-espec?ficos DAPICMA3, enzimas de restri??o AluI, EcoRI, TaqI, PstI e HinfI e pela hibrida??o in situ com sondas de DNA ribossomal 18S e 5S. A ultra-estrutura heterocromat?nica de A. quadricornis e A. polygonius, revelaram um marcante conte?do heterocrom?tico, situa??o que pode indicar que o ac?mulo ou perda de extenso conte?do de heterocromatinas poderiam ser respons?veis pelas extensas varia??es no conte?do gen?mico exibidas nas diferentes fam?lias dos Tetraodontiformes. As esp?cies Cantherhines macrocerus, C. pullus (Monacanthidae) e Melichthys niger (Balistidae) apresentam uma grande similaridade cariot?pica, tanto num?rica, como estruturalmente. Lagocephaluslaevigatus mostrou caracter?sticas citogeneticas similares (2n=44 e RONs simples) as esp?cies do g?nero Takifugu, o que refor?a a id?ia de seu relacionamento filogen?tico, e Colomesus psittacus apresentou o maior n?mero dipl?ide descrito para a fam?lia (2n=56) e grande quantidade de HC, caracter?sticas que o relacionariam com a fam?lia irm? Diodontidae. An?lises citogen?ticas em C. figueiredoi revelaram polimorfismos heterocrom?ticos, RONs m?ltiplas e cromossomos Bs, sendo estes eventos raros para peixes marinhos, estando possivelmente associados ? marcante reestrutura??o e redu??o gen?mica que esta fam?lia sofreu. Estas caracter?sticas, somadas aos dados morfol?gicos e moleculares sugerem que estas esp?cies compartilham de um mesmo ramo ancestral, com poss?vel origem monofil?tica. Neste trabalho novas contribui??es ao conhecimento dos padr?es evolutivos enfrentados pelos Tetraodontiformes s?o fornecidas e discutidas sob perspectivas citotax?nomicas, gen?micas e evolutivas.