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Made available in DSpace on 2014-06-11T19:25:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-27Bitstream added on 2014-06-13T19:11:54Z : No. of bitstreams: 1 lopes_at_me_jabo.pdf: 1243839 bytes, checksum: 8708fa8a3a2a924bf7100eb2b3992e01 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A família Chrysopidae, pertencente à ordem Neuroptera, é a mais numerosa, compreendendo mais de 1200 espécies válidas, distribuídas em várias regiões do mundo, principalmente na Neotropical. A classificação desta família em gêneros e espécies é baseada em estudos da morfologia externa e da genitália de machos e fêmeas adultos. No entanto tem-se buscado novas metodologias para auxiliar tanto na taxonomia, quanto no conhecimento da evolução. A citogenética é uma ferramenta bastante importante para o conhecimento da extensão da biodiversidade, pois a partir de diferenças presentes nos cromossomos podem-se distinguir grupos, bem como padrões de evolução. Assim, o objetivo deste estudo foi caracterizar citogeneticamente 18 espécies de crisopídeos de ocorrência em Jaboticabal – SP., quanto ao número diplóide, morfologia e tamanho dos cromossomos, sistema cromossômico sexual, e quando possível, analisar o comportamento meiótico. Os cromossomos foram obtidos a partir de preparações gonadais masculinas e femininas de indivíduos adultos e de embriões, submetidos à coloração convencional e fotografados com câmara MOTICAM acoplada a microscópio e software IMAGE da Motic. Das 18 espécies analisadas, somente em Chrysoperla defreitasi não foi possível observar o par heteromórfico, no entanto as demais espécies exibiram sistema sexual do tipo XY/XX. O número diplóide variou entre 2n=16 nas espécies do gênero Leucochrysa a 2n=6 em Chrysopodes delicata, e esta diferença refletiu na morfologia cromossômica e permitiu diferenciar algumas das espécies. Nas espécies em que foram obtidas células meióticas observou-se a formação de bivalentes autossômicos com quiasmas terminais e univalentes sexuais assinápticos e aquiasmáticos / The family Chrysopidae, belonging to the Neuropteran order, is the most numerous, comprising more than 1200 valid species, distributed in various regions of the world, mainly in the Neotropical. The classification of this family into genera and species is based on the study of external morphology and male and female adults genitalia. However, new methods to assist the taxonomy and the evolution have been studied. The cytogenetic is a very important tool for to understand the extent of biodiversity, because the differences in the chromosomes can be distinguished group, and pattern of evolution. The objective of this study was to perform cytogenetic analysis of 18 species of green lacewings of occurrence in the city of Jaboticabal – SP, characterizing them as to the diploid number, shape and size of chromosomes, and when was possible, analyzed the meiotic behavior. The chromosomes preparations were obtained from adults male and female gonodals and embryos that were conventional stained and photographed with a MOTIC camera coupled in a microscope and software IMAGE from Motic. Of the 18 species analyzed, only Chrysoperla defreitasi was not possible to observe the heteromorphic pair, however the other species exhibited sexual system of the type XY / XX. The diploid number varied from 2n = 16 in the genus Leucochrysa to 2n = 6 in Chrysopodes delicata, and this difference reflected in chromosome morphology and allowed to differentiate some of the species. In the species in which meiotic cells had obtained it was possible observed the formation of autosomal bivalents with terminal chiasmata and univalents sex without chiasmata and synaptic

by Lai, Shiu Hong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 129-147). / TITLE PAGE --- p.I / THESIS COMMITTEE --- p.II / ABSTRACT --- p.III / ACKNOWLEDGMENTS --- p.V / ABBREVIATIONS --- p.VI / TABLE OF CONTENTS --- p.VII / LIST OF TABLES --- p.XI / LIST OF FIGURES --- p.XII / Chapter Chapter 1. --- Arbitrarily Primed Polymerase Chain Reaction (AP-PCR) Analysis of Lent inula edodes / Chapter 1. --- Introduction / Chapter 1.1 --- General Introduction --- p.1 / Chapter 1.2 --- Purpose of Study --- p.4 / Chapter 2. --- Literature Review / Chapter 2.1 --- Biology of Lentinula edodes / Chapter 2.1.1 --- "Overview," --- p.6 / Chapter 2.1.2 --- Life Cycle of Lentinula edodes --- p.6 / Chapter 2.1.3 --- Dedikaryotization (Monokaryotization) --- p.12 / Chapter 2.2 --- Genome Analysis of the Mushroom --- p.13 / Chapter 2.3 --- Genetic Markers of Lentinula edodes / Chapter 2.3.1 --- Overview --- p.15 / Chapter 2.3.2 --- Auxotrophic Markers --- p.16 / Chapter 2.3.3 --- Biochemical Markers --- p.17 / Chapter 2.3.4 --- Molecular Markers / Chapter 2.3.4.1 --- RFLPs --- p.19 / Chapter 2.3.4.2 --- PCR-Based Markers --- p.20 / Chapter 2.4 --- Polymerase Chain Reaction (PCR) / Chapter 2.4.1 --- The principle of PCR --- p.22 / Chapter 2.4.2 --- Applications of PCR on Mushroom Studies --- p.26 / Chapter 2.5 --- Arbitrarily Primed Polymerase Chain Reaction / Chapter 2.5.1 --- Principle of AP-PCR --- p.27 / Chapter 2.5.2 --- Applications of AP-PCR on Mushroom Studies --- p.29 / Chapter 2.6 --- Genetic Linkage Analysis / Chapter 2.6.1 --- Overview --- p.31 / Chapter 2.6.2 --- The LOD Score Method --- p.34 / Chapter 3. --- Materials and Methods / Chapter 3.1 --- Mushroom Strains and Culture Media --- p.36 / Chapter 3.2 --- Culture Method --- p.36 / Chapter 3.3 --- Solutions --- p.36 / Chapter 3.4 --- Primers --- p.38 / Chapter 3.5 --- Isolation of DNA from Lentinula edodes / Chapter 3.5.1 --- Mini-Preparation of Fungal DNA from L. edodes for PCR amplification --- p.42 / Chapter 3.5.2 --- Cesium Chloride Method: Mini-Preparation of Fungal DNA for PCR amplification --- p.43 / Chapter 3.6 --- Quantitative Measurements of DNA --- p.44 / Chapter 3.7 --- Arbitrarily Primed Polymerase Chain Reaction (AP-PCR) for the Amplification of Genomic DNA of L. edodes --- p.45 / Chapter 3.8 --- Analysis of DNA Samples with Agarose Gel Electrophoresis --- p.46 / Chapter 3.9 --- Analysis of DNA Samples with Polyacrylamide Gel Electrophoresis (PAGE) --- p.47 / Chapter 3.10 --- Silver Staining --- p.48 / Chapter 3.11 --- Single Stranded Conformation Polymorphism (SSCP) Analysis of Polymorphic DNA Fragments / Chapter 3.11.1 --- Elution and Amplification of DNA --- p.49 / Chapter 3.11.2 --- PCR-SSCP --- p.50 / Chapter 3.12 --- Segregation and Linkage Analysis / Chapter 3.12.1 --- Chi-Square Test --- p.51 / Chapter 3.12.2 --- The LOD Score Method --- p.52 / Chapter 4. --- Results / Chapter 4.1 --- DNA Extraction --- p.54 / Chapter 4.2 --- AP-PCR Amplified Fragments and Fragment Number --- p.58 / Chapter 4.3 --- Dedikaryotization Demonstration --- p.60 / Chapter 4.4 --- Identification of Polymorphic Genetic Markers --- p.64 / Chapter 4.4.1 --- AP-PCR Fingerprints from Single Primer --- p.66 / Chapter 4.4.2 --- AP-PCR Fingerprints Using Two Primers --- p.76 / Chapter 4.5 --- Segregation of Polymorphic Markers in Single Spore Isolates (SSIs) --- p.81 / Chapter 4.6 --- Single Stranded Conformation Polymorphism (SSCP) of Identified Polymorphic DNA Fragments --- p.86 / Chapter 4.7 --- Linkage Analysis of the Identified AP-PCR Markers --- p.89 / Chapter 5. --- Discussions / Chapter 5.1 --- DNA Extraction --- p.92 / Chapter 5.2 --- Arbitrary Primers --- p.93 / Chapter 5.3 --- Dedikaryotization Demonstration --- p.95 / Chapter 5.4 --- Identification of Polymorphic Genetic Markers --- p.96 / Chapter 5.5 --- AP-PCR Analysis of a Mushroom --- p.97 / Chapter 5.6 --- Mendelian Segregation Pattern of the Polymorphic Markers --- p.99 / Chapter 6. --- Conclusion and Further Studies --- p.101 / Chapter 2. Electrophoretic Karyotype Analysis of Lentinula edodes / Chapter 7. --- Introduction --- p.106 / Chapter 8. --- Literature Review / Chapter 8.1 --- Overview --- p.108 / Chapter 8.2 --- Protoplasts --- p.109 / Chapter 8.3 --- Pulsed Field Gel Electrophoresis (PFGE) / Chapter 8.3.1 --- Principle --- p.110 / Chapter 8.3.2 --- Applications of PFGE in Studies of Fungi --- p.112 / Chapter 9. --- Materials and Methods / Chapter 9.1 --- Strains and Culture Media --- p.114 / Chapter 9.2 --- Solutions --- p.114 / Chapter 9.3 --- Production of Lentinula edodes Protoplast --- p.115 / Chapter 9.4 --- Electrophoretic Conditions --- p.116 / Chapter 9.4.1 --- Condition for Saccharomyces cerevisiae chromosomes --- p.117 / Chapter 9.4.2 --- Condition for Candida albicans chromosomes --- p.117 / Chapter 9.4.3 --- Condition for Schizosaccharomyces pombe chromosomes --- p.118 / Chapter 10. --- Results / Chapter 10.1 --- Protoplast Production of Lentinula edodes / Chapter 10.1.1 --- Effects of Age of Mycelium on Protoplast Yield --- p.119 / Chapter 10.1.2 --- Effects of Various Osmotic Stabilizers on Protoplast Yield --- p.121 / Chapter 10.1.3 --- Effects of Two Lytic Enzymes on Protoplast Yield --- p.123 / Chapter 10.1.4 --- The Optimal Condition --- p.123 / Chapter 10.2 --- Electrophoretic Karyotype of L. edodes --- p.124 / Chapter 11. --- Discussions / Chapter 11.1 --- Protoplast Production of Lentinula edodes --- p.126 / Chapter 11.2 --- Electrophoretic Karyotype --- p.127 / REFERENCES

Jiang Qing Lan. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (p. 171-188). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Orientador: Eliana Regina Forni Martins / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-11T11:47:41Z (GMT). No. of bitstreams: 1 Oliveira_VanessaMancusode_M.pdf: 2101182 bytes, checksum: 4b6aba0f8011aa1dbf7656da7b98141b (MD5) Previous issue date: 2008 / Resumo: O gênero Vernonia é o maior da tribo Vernonieae (Asteraceae), possuindo mais de 1.000 espécies. O Brasil é o maior centro de diversidade das espécies do Novo Mundo deste gênero. As subdivisões de Vernonia têm sido de difícil circunscrição devido ao seu tamanho, que acomoda muitas variações e paralelismos. Recentemente, este gênero foi segregado em outros 22, e o mesmo ficou restrito apenas aos representantes da América do Norte. Entretanto, essa mudança não foi aceita por alguns autores. O objetivo deste trabalho foi subsidiar a proposta sobre a segregação de Vernonia em gêneros menores (sensu ROBINSON) ou da manutenção de sua integridade (sensu BAKER) mediante a comparação de cariótipo. No total, foram estudadas 14 espécies de Vernonia. Oito delas, pertencentes à seção Lepidaploa, correspondentes às subseções Axilliflorae, Macrocephalae, Oligocephalae, Paniculatae e Scorpioideae foram estudadas através da técnica de Giemsa. As espécies foram coletadas em áreas de cerrado e de campo rupestre e em ambiente perturbado, nos Estados de São Paulo, Minas Gerais e Goiás. Foram realizadas contagens cromossômicas nestas mesma espécies, que variaram de 2n=20 a 2n=60 e, elaborados cariótipos, verificando-se o predomínio de cromossomos metacêntricos, e alguns submetacêntricos. O tamanho dos cromossomos variou de 0,73 a 3,5µm, o tamanho total de cromatina (CTC) de 23,5 a 44,9 µm e, o índice de assimetria TF% de 32,2 a 45,9. O índice de assimetria intracromossômica (A1) variou de 0,30 a 0,85, enquanto o índice de assimetria intercromossômica (A2) de 0,14 a 0,40. Vernonia rubriramea foi a espécie que mostrou ter cariótipo mais simétrico. Também foi elaborada uma coletânea dos números cromossômicos das espécies de Vernonia, incluindo os resultados obtidos e os disponíveis em literatura, como publicações de revisão e artigos específicos. Foram aplicadas as técnicas de bandamentos AgNOR e CMA/DA/DAPI e a técnica de FISH com a seqüência de DNAr 45S em algumas espécies de Vernonia, incluindo também algumas que tiveram seu cariótipo elaborado com técnicas de coloração convencional (Giemsa). De modo geral, as espécies apresentaram dois sítios de DNAr 45S terminais, sempre localizados no braço curto do cromossomo, com exceção de V. condensata e V. geminata, com quatro, e V. bardanoides, com seis sítios. A hibridação in situ evidenciou, na população de V. geminata coletada em Assis, um par de sítios de DNAr 45S centromérico, e na população coletada em Analândia, dois sítios apareceram em cromossomos B. Foram observados até seis cromossomos Bs nesta última população. Essa foi a única espécie que apresentou cromossomos extranumerários. Os bandamentos CMA/DA/DAPI e AgNOR evidenciaram em algumas espécies, um par de bandas CMA+ e um par de bandas NOR, sempre localizadas na região terminal do braço curto dos cromossomos, com exceção de V. platensis e V. scorpioides, que apresentaram três pares de bandas CMA+. Os dados cariotípicos obtidos no presente trabalho e mais dados em literatura não são suficientes para apoiar conclusivamente qualquer das propostas taxonômicas vigentes para Vernonia, devido à inexistência de um padrão cariotípico característico/distintivo para cada grupo taxonômico, ou seja, para suas seções e subseções (sensu BAKER) ou para os novos gêneros (sensu ROBINSON), considerados a partir de seu desmembramento. No entanto, até o momento, parece existir uma tênue relação com a conceituação de ROBINSON (1999a) para os gêneros Lessingianthus, Vernonanthura, e Chrysolaena, com os números cromossômicos obtidos. Diante da não disponibilidade de sondas funcionais com as seqüências de DNAr 5S e DNA telomérico, tentou-se a obtenção de sondas específicas para Vernonia mediante a técnica de PCR com primers específicos. Obteve-se sucesso apenas na amplificação do DNA telomérico com os primers de Arabidopsis (Tel-1 e Tel-2) / Abstract: The genus Vernonia is the largest of the tribe Vernonieae (Asteraceae), comprising more than 1.000 species. The greatest center of diversity of this genus from the New World is in Brazil. The subdivisions of Vernonia have been a difficult constituency because of its size, which accommodates many variations and parallels. Recently, this genus was dismembered into 22 genera and Vernonia was restricted to North America. However, most of the modifications proposed were not accepted for others workers in this field. In order to assess the validity of maintaining this genus (sensu BAKER) or dividing it into several lesser genera (sensu ROBINSON), we described a mitotic analyses (Giemsa technique) of eight species of Vernonia, belonging to subsections Axilliflorae, Macrocephalae, Oligocephalae, Paniculatae and Scorpioideae of the section Lepidaploa. Specimens were collected in ¿cerrado¿, rupiculous and disturbed areas, in the states of Sao Paulo, Minas Gerais and Goiás. Chromosome numbers (2n=20 to 2n=60) and karyotypes were analyzed, with predominance of metacentric and some submetacentric chromosomes. The chromosomes size varied from 0.73 to 3.5µm, the total chromatin length (TCL) ranged from 23.5 to 44.9µm, and the asymmetry index TF% ranged from 32.2 to 45.9%. The intrachromosomal asymmetry index (A1) varied from 0.30 to 0.85, while the interchromosomal asymmetry index (A2) ranged from 0.14 to 0.40. The species V. rubriramea had the most symmetrical karyotype. We prepared a compilation of the chromosome numbers of species of Vernonia, including the results obtained here and the available literature, as publications of review and specific articles. We applied AgNOR and CMA/DA/DAPI banding and FISH with the sequence of rDNA 45S in some species of Vernonia, including some that had their karyotype analyzed with the Giemsa technique. The chromosome number ranged from 2n = 20 to 60, but most frequent chromosomal numbers were 2n = 32 and 34. Generally, the species showed two terminals sites of rDNA 45S, always located on the short arm of chromosome, except for V. condensata and V. geminata, with four, and V. bardanoides, with six sites. The technique of FISH showed in the population of V. geminata collected in Assis, one pair of centromeric sites of rDNA 45S, and the population collected in Analândia, two sites appeared in B chromosomes. That was the only species that showed extra numerous chromosomes. The CMA/DA/DAPI and AgNOR banding neither evidenced in some species, one pair of CMA+ bands and one pair of NOR bands, always located in the terminal region of the short arm of chromosome, except for V. platensis and V. scorpioides, which had three pairs of CMA+ bands. Despite of the little representativity of the samples, the karyotypic characters obtained in this study and in literature did not allow conclusive support to the taxonomic proposes to Vernonia, due to the inexistence of a distinctive/characteristic karyotypic pattern for each taxonomic group, which means, sections and subsections (sensu BAKER) or new genera (sensu ROBINSON). Nevertheless, the available data indicate only a tenuous relationship between the chromosome numbers observed here and reported in the literature compared to the taxonomic reorganization of the genera Lessingianthus, Vernonanthura and Chrysolaena. Due to the non-availability of functional probes with the rDNA 5S and telomeric sequences, we tried to obtain probes specific for Vernonia with the PCR technique with specific primers. We had sucess only in the DNA amplification with telomeric primers of Arabidopsis (Tel-1 and Tel-2) / Mestrado / Doutor em Biologia Vegetal

Orientadores: Shirlei Maria Recco-Pimentel, Ana Cristina Prado Veiga-Menoncello / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-12T04:20:05Z (GMT). No. of bitstreams: 1 Favero_EduardoRondelli_M.pdf: 1364067 bytes, checksum: 03d34437fbe08a46a8cdd3631aaedcc9 (MD5) Previous issue date: 2008 / Resumo: O gênero Pseudopaludicola, pertencente à família Leiuperidae, compreende atualmente 12 espécies de rãs de pequeno tamanho, distribuídas pela América do Sul, sendo a ocorrência de oito delas foi relatada para o Brasil. Devido à grande semelhança morfológica entre espécies, algumas ocorrendo em simpatria, confusões taxonômicas são freqüentes. Alguns estudos morfológicos acerca deste gênero foram realizados, mas as relações de parentesco inter- e intragenéricas de Pseudopaludicola permanecem pouco esclarecidas. As poucas informações citogenéticas para o gênero Pseudopaludicola restringiam-se apenas à determinação do número de cromossomos e análise do cariótipo por métodos de coloração convencional. Para Pseudopaludicola falcipes, em especial, foi descrita uma variação intra-específica do número de cromossomos, de 2n=16 à 2n=20. O presente estudo visa contribuir com dados citogenéticos para a caracterização de espécies de Pseudopaludicola e para o entendimento dos processos envolvidos na evolução cariotípica do gênero. Foram analisados os cariótipos de exemplares de Pseudopaludicola falcipes, P. ameghini (sensu Cope, 1887) e P. mystacalis de suas respectivas localidades-tipo (região de Porto Alegre, RS e Chapada dos Guimarães, MT), de P. mystacalis e de P. ternetzi, de Uberlândia (MG), de Pseudopaludicola aff. falcipes I, II, III e IV, da região noroeste do estado de São Paulo (municípios de Santa Fé do Sul, Vitória Brasil, Palestina e Icém), de Pseudopaludicola aff. mystacalis I, II, III e IV, dos municípios de Icém (SP), Barreirinhas (MA) e Urbano Santos (MA) e Pseudopaludicola sp. 1, 2 e 3, sendo as duas primeiras provenientes de Poconé (MT) e a terceira de Santa Terezinha (MT). As metáfases foram obtidas de suspensões de células de epitélio intestinal e testículo, e coradas com Giemsa ou submetidas às técnicas impregnação por prata (Ag-NOR) para detecção de NOR e de bandamento C, para a localização de heterocromatina. Os dados obtidos revelaram uma variação interespecífica quanto ao número de cromossomos. Dentre os espécimes provenientes de Poconé, MT, havia dois cariótipos distintos, com 2n=22 e com 2n=16 cromossomos (Pseudopaludicola sp. 1 e 2) e os de Icém, SP, com indivíduos 2n=20 e 2n=16 cromossomos (Pseudopaludicola aff. mystacalis, respectivamente I e II). Pseudopaludicola falcipes e Pseudopaludicola sp.1, de Poconé, apresentaram 2n=22 e a NOR localizada na região pericentromérica do braço longo do par 8. Estas espécies diferiram, na morfologia da NOR, sendo heteromórfica em P. falcipes e homomórfica em Pseudopaludicola sp. 1, e na localização de algumas bandas heterocromáticas. Pseudopaludicola ameghini (sensu Cope, 1887), P. ternetzi e Pseudopaludicola aff. mystacalis I de Icém apresentaram 2n=20 cromossomos e a NOR localizada na região telomérica do braço longo do par 9. O cariótipo de P. ternetzi diferiu do de P. ameghini tanto pela classificação morfológica distinta do par 7 quanto pelo padrão de distribuição de heterocromatina. Pseudopaludicola mystacalis, bem como todos os espécimes de Pseudopaludicola aff. falcipes I, II, III e IV, Pseudopaludicola aff. mystacalis II, III e IV e Pseudopaludicola sp. 2 e 3 apresentaram 2n=16 cromossomos metacêntricos e submetacêntricos, com a NOR localizada na região pericentromérica do braço curto do par 4. Vários espécimes apresentaram um heteromorfismo de tamanho em relação aos homólogos do par 4 (morfo 4 e morfo 4'), alterando a classificação desse cromossomo para metacêntrico em algumas populações. Em P. mystacalis, P. aff. falcipes I, II, III e IV, P. aff. mystacalis II, III e IV e Pseudopaludicola sp 2 e 3 foram detectados blocos de heterocromatina fortemente marcados nas regiões pericentroméricas no braço curto do par 1 e longo do par 2. Os resultados obtidos mostram que P. ameghini (sensu Cope, 1887) com 2n=20 e P. mystacalis com 2n=16, são unidades taxonômicas distintas e que os espécimes tidos como Pseudopaludicola aff. falcipes) e como Pseudopaludicola sp. mostraram-se citogeneticamente relacionados à P. mystacalis e não à P. falcipes que possui 2n=22 cromossomos. Desta forma, os nossos dados sugerem a retirada de P. ameghini da sinonímia de P. mystacalis e reforçam a necessidade de uma revisão taxonômica no gênero. / Abstract: The genus Pseudopaludicola (family Leiuperidae) comprises 12 species of small sized frogs, which are widely distributed in South America. In Brazil, eight species are described within this genus, and several of them are sympatric. The relevant morphological similarities among the Pseudopaludicola species have contributed to the still poor understanding of many aspects of their taxonomy, including inter- and intrageneric relationships. Cytogenetic data on Pseudopaludicola have been restricted to karyotype analyses using conventional Giemsa staining. Variation in intraspecific chromosomal number was described in P. falcipes, ranging from 2n=16 to 2n=20. In the present work, Brazilian Pseudopaludicola species were submitted to cytogenetic analysis aiming at their further characterization and attempting to better understanding the karyotypical evolution of this genus. The analyzed species were Pseudopaludicola falcipes (Porto Alegre, RS), P. ameghini (sensu Cope, 1887) and P. mystacalis (Chapada dos Guimarães, MT), P. mystacalis and P. ternetzi (Uberlândia, MG), P. aff. falcipes I, II, III and IV (respectively from Santa Fé do Sul, Vitória Brasil, Palestina and Icém, SP), P. aff. mystacalis I, II, III and IV (Icém, SP, Barreirinhas, MA, and Urbano Santos, MA), and Pseudopaludicola sp. 1 and sp. 2 (Poconé, MT) and sp. 3 (Santa Terezinha, MT). Metaphases were obtained from suspensions of intestinal epithelium and testicular cells, and stained with Giemsa or submitted to silver staining technique in order to detect the nucleolus organizing regions (Ag-NOR), and C-banding, for heterochromatin localization. The results revealed interspecific chromosomal number variation. In the Pseudopaludicola sp. 1 and 2 specimens (Poconé MT), two distinct karyotypes were identified, respectively with 2n=22 and 2n=16 chromosomes. Within the Pseudopaludicola aff. mystacalis from Icém, SP, the analyzed specimens had 2n=20 and 2n=16 chromosomes, being nominated I and II, respectively. These data clearly indicated two criptic species of Pseudopaludicola within each of those two localities. The P. falcipes and Pseudopaludicola sp.1 (Poconé, MT) had 2n=22 and the NOR was located at the pericentromeric region in the long arm of the pair 8. The species differed in the NOR morphology, which was heteromorphic in P. falcipes and homomorphic in Pseudopaludicola sp.1, as well as in the localization of some C-bands. Pseudopaludicola ameghini (sensu Cope, 1887), P. ternetzi and Pseudopaludicola aff. mystacalis I (Icém, SP) had 2n=20 chromosomes and the NOR was located on the telomeric region in the long arm of the pair 9. The karyotypes of P. ternetzi and P. ameghini differed in the pair 7 morphology and in the heterochromatin distribution pattern. All analyzed specimens of P. mystacalis, P. aff. falcipes I, II, III and IV, P. aff. mystacalis II, III and IV, and Pseudopaludicola sp. 2 and 3, showed 2n=16 chromosomes, which were all metacentric and submetacentric, with the NOR located in the pericentromeric region of the short arm of the pair 4. In several of those specimens, the size heteromorphism of the pair 4 altered, from submetacentric to metacentric, the classification of one of the homologous of that pair. Strong pericentromeric C-bands were detected on the short arm of the pair 1 and on the long arm of the pair 2 in P. mystacalis, P. aff. falcipes I, II, III and IV, P. aff. mystacalis II, III and IV, and Pseudopaludicola sp. 2 and 3. As based on the cytogenetic data, P. ameghini (sensu Cope, 1887), with 2n = 20, and P. mystacalis, with 2n = 16, are distinct taxonomic units, and the specimens formerly identified as P. aff. falcipes and as Pseudopaludicola sp. were indeed cytogenetically closely related to P. mystacalis and not to P. falcipes, which has 2n = 22 chromosomes. Hence, our data suggest that P. ameghini is not a P. mystacalis synonymy and emphasize the importance of a taxonomic review of the genus Pseudopaludicola. / Mestrado / Biologia Celular / Mestre em Biologia Celular e Estrutural

The origin and geographical distribution of mammalian species (my examples are lemurs and bats) correlate with predictable chromosomal structural changes (KFT=karyotypic fission theory). Chromosome studies provide information about fertility between individuals and they are significant for identification of the geographical origin of reproductive isolation within mammal families. Each family predictably has chromosome sets with numbers that range from one to double the lowest number of chromosomes. The chromosome numbers of all species within a single family are used to reconstruct that family’s evolutionary geographical dispersion. Polymorphic chromosome numbers (that is a range such as 34, 35, 36, 37, and 38) in a single population indicate the location where chromosomal diversification arose. Chromosome numbers of descending order correlate with relative distance from fission epicenters as the fissioned chromosomes gradually spread to neighboring populations. Furthermore, the location of chromosomal diversification (that is “karyotypic fission events) is associated with geographical “zones of transition” (after Professor R.W. Wilkie). My analysis, mapped one (Lepilemuridae) of the five families of lemurs (Class Mammalia, Order Primates, sub-order Lemuridae). The origin of this family’s diversification is here hypothesized to have occurred at an ecological transition zone in Northern Madagascar between a humid evergreen-forest that extends to the East relative to a dry deciduous forest along the West Coast. My analysis of Vespertilionidae (insectivorous bats representing one third of all bat species) suggests a diversification event occurred in Asia; South China. Geographical distribution is important in the formation of biological diversity. A single species can inhabit a wide range and exhibit great diversity that is brought about by natural selection. The Holarctic reindeer found in Scandinavia, Russia, China, Canada and Alaska (including caribou) are all a single species Rangifertarandus that exhibits variation in size and in coat pattern, changes brought about by adaptive selection by the environment or human selective breeding but they all have 70 similar chromosomes and they are all reproductively compatible. There is a single species of reindeer. Although, there is measurable DNA sequence divergence; there has been no “speciation” as these circumpolar cervids are genetically compatible.

Orientador: Luciana Bolsoni Lourenço / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-26T03:48:33Z (GMT). No. of bitstreams: 1 Nascimento_Juliana_D.pdf: 2106853 bytes, checksum: f64fce074a208a02085fbde516c48f9c (MD5) Previous issue date: 2014 / Resumo: Recentes análises filogenéticas e citogenéticas têm revelado grande diversidade dentre leiuperídeos do grupo Physalaemus cuvieri. O parafiletismo de P. cuvieri em relação a Physalaemus ephippifer foi inferido com base na análise de sequências de DNA, e linhagens reconhecidas dentre indivíduos de P. cuvieri parecem representar espécies ainda não descritas. A espécie Physalaemus ephippifer ocorre na foz do Rio Amazonas no Brasil e possivelmente na Guiana, mas sua correta distribuição geográfica permanece desconhecida. Análises citogenéticas de indivíduos de P. ephippifer da localidade-tipo (Belém-PA) revelaram um interessante heteromorfismo entre os homólogos do par cromossômico 8 de fêmeas, caracterizando a presença de um sistema de determinação sexual cromossômico ZZ/ZW. Os cromossomos Z e W apresentam uma NOR distal no braço longo, adjacente a uma banda heterocromática terminal, e o cromossomo W difere do Z por apresentar um segmento terminal no braço curto que inclui uma NOR e uma banda heterocromática. As NORs de P. ephippifer se mostraram co-localizadas, em experimentos de hibridação in situ, com as sequências de DNA repetitivo Pep194. Com o objetivo de contribuir para a investigação da diversidade genética e citogenética de populações amazônicas de Physalaemus do grupo P. cuvieri, analisamos as relações filogenéticas e os cariótipos de indivíduos de P. ephippifer de Santa Isabel-PA e exemplares coletados em localidades do Pará (Monte Alegre, Óbidos, Alenquer, Curuá, Parauapebas e Prainha) e de Roraima (Viruá), cuja identificação taxonômica é incerta. Para tanto, as preparações cromossômicas obtidas de machos a partir de suspensões de células do intestino e testículo foram submetidas à coloração convencional por Giemsa, bandamento C, coloração com DAPI, impregnação por prata e experimentos de hibridação in situ com sondas de DNA repetitivo Pep194 e DNA ribossomal 28S. Além disso, utilizamos uma sonda, chamada de Zqter, produzida a partir de um segmento microdissecado da região terminal do braço longo do cromossomo Z de P. ephippifer. As relações filogenéticas foram inferidas com base em sequências de DNA mitocondrial dos genes 12S, RNAt-val e 16S, utilizando o critério de máxima parcimônia. Os cariótipos dos indivíduos de Physalaemus sp. em análise foram semelhantes entre si, porém três citótipos foram reconhecidos devido ao padrão diferencial na distribuição das NORs no par 8. Os cariótipos dos espécimes de Monte Alegre, Óbidos, Alenquer, Prainha e Curuá (Citótipo I) e Parque Nacional do Viruá (Citótipo II) mostraram uma NOR pericentromérica no braço curto do par 8, porém apenas o Citótipo I foi portador de uma NOR terminal no braço longo desse mesmo par. O exemplar de Parauapebas (Citótipo III) mostrou duas NORs no braço longo dos cromossomos do par 8. Nas inferências filogenéticas, os exemplares de Physalaemus sp. portadores dos Citótipos I e II foram recuperados no mesmo clado (Clado A). Neste clado, foi possível observar uma subestruturação que acompanha a variação cariotípica detectada, no entanto a distância genética entre os dois grupos em questão, inferida com base em sequências parciais do gene 16S, foi baixa (0,1%). É provável, portanto, que os dois citótipos representem uma variação intraespecífica. O exemplar de Parauapebas incluído nas análises filogenéticas (Clado B) apresentou-se como grupo-irmão de um clado que incluiu P. ephippifer (Clado C) e o clado correspondente a uma das linhagens de P. cuvieri (Clado D, equivalente à Linhagem 1 de P. cuvieri, que agrupa indivíduos do norte e do nordeste do Brasil). A alta distância genética do Clado A em relação aos Clados B-D, inferida com base em segmento do gene 16S, sugere que o Clado A possa representar uma espécie ainda não descrita. Nas metáfases de indivíduos de Viruá, Alenquer e Curuá a sonda Zqter foi detectada na região pericentromérica do braço curto do par 8, adjacente às NORs detectadas pela sonda Pep194, sugerindo que esta região é homeóloga a região terminal ao braço longo do Z de P. ephippifer. As homelogias encontradas sugerem um possível evento de inversão cromossômica durante diferenciação desses cariótipos. A sonda Pep194 detectou, além das NORs dos cromossomos Z e W de P. ephippifer e da NOR pericentromérica de Physalaemus sp., as NORs dos cromossomos 8 e 9 de indivíduos de P. cuvieri de Urbano Santos-MA e os cromossomos 8 e 11 de indivíduos de P. cuvieri de Palestina-SP. Em contraste, essa sonda não detectou a NOR pericentromérica do cromossomo 9 de P. centralis, uma espécie mais distantemente relacionada filogeneticamente a P. cuvieri e P. ephippifer / Abstract: Recent phylogenetic and cytogenetic analyses have revealed high diversity among leiuperids of the Physalaemus cuvieri group. The paraphyly of P. cuvieri with respect to Physalaemus ephippifer was inferred based on the analysis of DNA sequences, and genetic lineages that included individuals at first identified as P. cuvieri may represent undescribed species. P. ephippifer occurs at the mouth of the Amazon River in Brazil and possibly in Guyana, but its geographical distribution is unclear. Cytogenetic analyses of individuals of P. ephippifer from the type locality (Belém-PA) revealed an interesting heteromorphism between the homologous chromosomes of pair 8 in females, which characterized the presence of a chromosomal sex determination system ZZ/ZW. The Z and W chromosomes have a distal NOR in the long arm, adjacent to a terminal heterochromatic band, and the W chromosome differs from Z chromosome by a terminal segment in the short arm that comprises a NOR and a heterochromatic band. The NORs of P. ephippifer are shown co-located with DNA repetitive Pep194 in in situ hybridization experiments. Aiming to contribute to the investigation of the genetic and cytogenetic diversity of Amazonian populations belonging to P. cuvieri group, we analyzed the phylogenetic relationships and the karyotypes of individuals of P. ephippifer from Santa Isabel-PA and specimens collected from seven localities in the states of Pará (Monte Alegre, Óbidos, Alenquer, Curuá, Parauapebas and Prainha) and Roraima (Viruá), for which the taxonomic identification is uncertain. Chromosome preparations obtained from male individuals were subjected to conventional staining with Giemsa, C-banding, DAPI staining, silver impregnation and in situ hybridization with probes for the repetitive DNA Pep194 and ribosomal DNA 28S. In addition, we used a probe, named Zqter probe, produced from a segment microdissected from the terminal region of the long arm of the Z chromosome of P. ephippifer. Phylogenetic relationships were inferred based on mitochondrial DNA sequences of 12S, tRNA-val and 16S genes, using the criterion of maximum parsimony. The karyotypes of the individuals of Physalaemus sp. analyzed were similar, but three cytotypes were recognized with differential distribution of NORs in pair 8. Individuals from Monte Alegre, Óbidos, Alenquer, Prainha and Curuá (Cytotype I) showed a pericentromeric NOR in the short arm of pair 8 and a terminal NORin the long arm. In the male karyotypes of Viruá (Cytotype II), the NOR was located in the pericentromeric region of the short arm of pair 8. The individuals from Parauapebas (Cytotype III) showed two NORs in the long arm of chromosome pair 8. In the phylogenetic inferences, the specimens of Physalaemus sp. with Cytotypes I and II were nested in the same clade (Clade A). This clade showed a substructure that reflected the two karyotypic groups detected. The genetic distance between these two groups, however, inferred from partial sequences of the 16S gene, was low 0,1 %. It is likely, therefore, that the two cytotypes represent an intraspecific variation. The exemplar of Parauapebas included in the phylogenetic analysis (Clade B) was the sister group of a clade that included P. ephippifer (Clade C) and the clade corresponding to one of the lineages of P. cuvieri (Clade D, equivalent to Lineage 1 of P. cuvieri, which groups individuals of northern and northeastern Brazil). The high genetic distance between Clade A and clades B-D, inferred from 16S gene segment, suggests that Clade A may represent an undescribed species. In metaphases of individuals from Viruá, Alenquer and Curuá, the Zqter probe was detected in the pericentromeric region of the short arm of pair 8, adjacent to the NOR detected by the probe Pep194, suggesting that this region is homeologous with the terminal region of the long arm of the Z chromosome of P. ephippifer. The inferred homeologies suggest a possible event of chromosomal inversion along the differentiation of these karyotypes. The Pep194 probe detected, beyond the NORs of the Z and W chromosomes of P. ephippifer and the pericentromeric NOR of chromosomes 8 of the cytotypes of Physalaemus sp., the NORs of chromosomes 8 and 9 of individuals of P. cuvieri from Urbano Santos-MA and chromosomes 8 and 11 of individuals P. cuvieri from Palestina-SP. In contrast, this probe did not detect the pericentromeric NOR of chromosome 9 of P. centralis, which is distantly related phylogenetically to P. cuvieri and P. ephippifer / Doutorado / Biologia Celular / Doutora em Biologia Celular e Estrutural

The present thesis deals with the phenomenon of additional sex chromosomes in Cimex lectularius (Hemiptera: Heteroptera: Cimicidae) using genome size analysis combined with the classical cytogenetic approach. Also, five other cimicid species and 12 species from the family Nabidae were analysed identically for comparative purposes. The thesis also pursues a description of methodical approaches of cytogenetics and flow cytometry in the study of C. lectularius. Recently analysed European specimens of C. lectularius from human host exhibited 12 distinct cytotypes, with a variable number of chromosomes X from two to 20 (2n♂ = 26+X1X2Y to 26+X1-20+Y). The fragmentation hypothesis of C. lectularius additional chromosomes X origin was established in the second half of the 20th century. However, the present genome size measurements suggest that various chromosomal rearrangements as duplication or deletion besides the fragmentation could occur. Males with basic cytotype 2n = 26+X1X2Y had average genome size of 2C = 1.94 pg, in contrast male with 2n = 26+X1-7+Y yielded 2C = 2.26 pg and also specimens with genome size decrease 2C = 1.69 pg appeared. The most informative turned up to be the relative genome size of sperm cells n = 13+X1X2 and n = 13+Y, where specimens with higher chromosome number showed...

Phylogenetic relationships of a sample comprising 248 bats belonging to 19 species and four families (Hipposideridae, Rhinolophidae, Molossidae and Vespertilionidae) from Senegal (Western Africa) were investigated with the use of multi-locus sequence data and non- differentially stained chromosomes. The karyotypes of Hipposideros ruber, H. tephrus, H. jonesi and H. cyclops were described for the first time. The standard Hipposideros formula was recorded in H. tephrus, H. jonesi and H. ruber (2n = 32, FNa = 60, FN = 64). The karyotypes of H. cyclops (2n = 36, FN = 66) and H. gigas (2n = 52, FN = 64) substantially diverged from this typical chromosomal complement. Rhinolophus landeri and R. fumigatus shared the same diploid number (2n = 58), but differed in the chromosome morphology (R. fumigatus - FNa = 60, FN = 64; R. landeri - FNa = 64, FN = 68). Rhinolophus landeri was found karyotypically distinct to other African populations, thus signalling a possible presence of cryptic forms within this species. The karyotypes of Chaerephon pumilus and Mops condylurus had a 2n = 48, FN = 54 and were similar to other previously studied species of this chromosomally conservative family. Chromosomal, Bayesian, maximum likelihood and genetic distance analyses revealed an indication for the existence of cryptic...