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Evolução Cromossômica Em Plantas De Inselbergues Com Ênfase Na Família Apocynaceae Juss. Angeline Maria Da Silva Santos

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Evolução Cromossômica Em Plantas De Inselbergues Com Ênfase Na Família Apocynaceae Juss. Angeline Maria Da Silva Santos UNIVERSIDADE FEDERAL DA PARAÍBA CENTRO DE CIÊNCIAS AGRÁRIAS PÓS-GRADUAÇÃO EM AGRONOMIA CAMPUS II – AREIA-PB Evolução cromossômica em plantas de inselbergues com ênfase na família Apocynaceae Juss. Angeline Maria Da Silva Santos AREIA - PB AGOSTO 2017 UNIVERSIDADE FEDERAL DA PARAÍBA CENTRO DE CIÊNCIAS AGRÁRIAS PÓS-GRADUAÇÃO EM AGRONOMIA CAMPUS II – AREIA-PB Evolução cromossômica em plantas de inselbergues com ênfase na família Apocynaceae Juss. Angeline Maria Da Silva Santos Orientador: Prof. Dr. Leonardo Pessoa Felix Tese apresentada ao Programa de Pós-Graduação em Agronomia, Universidade Federal da Paraíba, Centro de Ciências Agrárias, Campus II Areia-PB, como parte integrante dos requisitos para obtenção do título de Doutor em Agronomia. AREIA - PB AGOSTO 2017 Catalogação na publicação Seção de Catalogação e Classificação S237e Santos, Angeline Maria da Silva. Evolução cromossômica em plantas de inselbergues com ênfase na família Apocynaceae Juss. / Angeline Maria da Silva Santos. - Areia, 2017. 137 f. : il. Orientação: Leonardo Pessoa Felix. Tese (Doutorado) - UFPB/CCA. 1. Afloramentos. 2. Angiospermas. 3. Citogenética. 4. CMA/DAPI. 5. Ploidia. I. Felix, Leonardo Pessoa. II. Título. UFPB/CCA-AREIA A Deus, pela presença em todos os momentos da minha vida, guiando-me a cada passo dado. À minha família Dedico esta conquista aos meus pais Maria Geovânia da Silva Santos e Antonio Belarmino dos Santos (In Memoriam), irmãos Aline Santos e Risomar Nascimento, tios Josimar e Evania Oliveira, primos Mayara Oliveira e Francisco Favaro, namorado José Lourivaldo pelo amor a mim concedido e por me proporcionarem paz na alma e felicidade na vida. Em especial à minha mãe e irmãos por terem me ensinado a descobrir o valor da disciplina, da persistência e da responsabilidade, indispensáveis para a construção e conquista do meu projeto de vida. E por dedicarem parte de suas vidas, na minha formação pessoal e profissional, por cumprirem este papel magistralmente e pelo amor intenso. Dedico AGRADECIMENTOS Ao Prof. Leonardo Pessoa Felix, por aceitar ter me orientado, pelos conhecimentos científicos, pela confiança, amizade e, acima de tudo, por ser um exemplo de profissional e pessoa que pretendo seguir. Ao Dr. Felipe Nollet Medeiros de Assis pelos conhecimentos, ajuda, por ser um profissional exemplar, amizade e por ter me acolhido, estando sempre à disposição. Ao Programa de Pós-Graduação em Agronomia, pela oportunidade de doutoramento. A Coordenação de Aperfeiçoamento de Pessoal de Nível Superior/CAPES, pelo financiamento desta pesquisa. A professora Drª Ana Emília Barros e Silva, Doutoras Fabiane Rabelo Costa e Lânia Isis Ferreira Alves pelas contribuições e sugestões dadas ao trabalho. A Alessandro Rapini e Roxana Barreto pela identificação das espécies de Apocynaceae e Commelinaceae. Aos colegas da Pós-Graduação em Agronomia pela parceira e ajuda mútua durante minha formação. Aos amigos conquistados ao longo da vida Patrick, Jackson, Filipe, Andreia, Magna, Carolline, Mariana, Edjane, Jeferson, Adeílson, Yovany e Tassinho. A Sarah Nascimento e Bruno César por terem se tornado mais que amigos, irmãos. Aos amigos do laboratório: Enoque, Cláudio, Joel, Amanda, Sibelle, Erton, Achilles, Saulo, Sócrates, Carolina, Larisse, Rodrigo, Felipe Coutinho, Mariana, Ednalva, Ingrid e Luciana. Obrigada pela amizade e companheirismo. Aos alunos e depois amigos conquistados durante o estágio de docência: David, Ulisses (In Memoriam), Jardel, Ingrid, Beatriz e Eduardo. A todos os professores que contribuíram decisivamente para a minha formação acadêmica, profissional, pessoal e pelos ensinamentos valorosos. Agradeço aos meus antigos professores, que me ensinaram com prazer e dedicação. As pessoas que de alguma forma passaram pela minha vida e contribuíram direta ou indiretamente para a concretização deste sonho e para a construção de quem sou hoje. Obrigada! SANTOS, Angeline Maria da Silva. D.S. Universidade Federal da Paraíba, agosto de 2017. Evolução cromossômica em plantas de inselbergues com ênfase na família Apocynaceae Juss. Leonardo Pessoa Felix. RESUMO Os inselbergues são caracterizados por apresentarem condições edafoclimáticas mais severas em relação ao ambiente do entorno, exercendo uma função seletiva importante para a evolução e especiação vegetal. A flora dos inselbergues pode apresentar diferenciação morfológica e cromossômica (níveis de ploidia), possibilitando a avaliação dos mecanismos envolvidos na diversificação das espécies. Uma hipótese longamente debatida sugere que os poliploides apresentam maior tolerância a condições ambientais mais extremas, quando comparadas com as suas contrapartes diploides. Sendo assim, o trabalho teve como objetivo analisar 28 espécies pertencentes a 10 diferentes famílias de angiospermas, ocorrentes em inselbergues na região Nordeste do Brasil com ênfase para a família Apocynaceae. Pontas de raízes foram pré-tratadas com 8HQ e Colchicina (apenas a espécie da família Costaceae) e fixadas em Carnoy. As lâminas foram preparadas pelo método de esmagamento em ácido acético e envelhecidas em temperatura ambiente. Em seguida, as melhores lâminas foram coradas com Chromomycin (CMA) e 4',6-Diamidino-2-Phenylindole (DAPI) e fotografadas. As espécies de Apocynaceae variaram de 2n = 18 a 2n = 34, com dados inéditos para seis espécies. Foram identificados três tipos de bandas heterocromáticas nas espécies da família Apocynaceae: bandas CMA+/DAPI−, CMA−/DAPI+ e CMA0/DAPI+, variando quanto ao padrão de distribuição, número e tamanho das bandas heterocromáticas. Bandas CMA+/DAPI− foram observadas em todas as espécies analisadas, e provavelmente é o tipo de heterocromatina associada as RONs. Outras espécies analisadas variaram de 2n = 18 para Chamaecostus subsessilis (Costaceae) até 2n = 76 em Dichorisandra hexandra (Commelinaceae). A poliploidia e a disploidia são os principais mecanismos de evolução cariotípica envolvidos na diversificação em Apocynaceae, sendo as espécies diploides as mais frequentemente encontradas em inselbergues. Dentre os gêneros analisados não se verificou uma relação clara entre número de bandas CMA+ e nível de ploidia. Das 27 espécies ocorrentes em inselbergues no nordeste do Brasil analisadas apenas sete são poliploides, ocorrendo especialmente nas famílias Apocynaceae, Commelinaceae, Euphorbiaceae e Melastomataceae. Além disso, não foi observada a ocorrência preferencial de citótipos poliploides em inselbergues para estas espécies, refutando a hipótese de que poliploides apresentam maior tolerância a condições edafoclimáticas mais extremas. Palavras-chave: Afloramentos, Angiospermas, Citogenética, CMA/DAPI, Ploidia SANTOS, Angeline Maria da Silva. D.S. Federal University of Paraiba, August of 2017. Chromosomal evolution in inselbergues plants with an emphasis on the family Apocynaceae Juss. Leonardo Pessoa Felix. ABSTRACT The inselbergs are characterized by presenting more severe edaphoclimatic conditions in relation to the surrounding environment, exerting an important selective function for the evolution and vegetal speciation. The flora of the inselbergs can present morphological and chromosomal differentiation (levels of ploidy), making possible the evaluation of the mechanisms involved in the diversification of the species. A long-debated hypothesis suggests that polyploids exhibit greater tolerance to more extreme environmental conditions when compared to their diploid counterparts. The objective of this work was to analyze 28 species belonging to 10 different families of angiosperms, occurring in inselbergues in the Northeast region of Brazil, with an emphasis on the Apocynaceae family. The root tips were pretreated with 8HQ and Colchicine (only the Costaceae family species) and fixed in Carnoy. The slides were prepared by the method of crushing in acetic acid and aged at room temperature. Then, the best slides were stained with chromomycin (CMA) and 4 ', 6-Diamidino-2-Phenylindole (DAPI) and photographed. The species of Apocynaceae ranged from 2n = 18 to 2n = 34, with new data for six species. Three types of heterochromatic bands were identified in family Apocynaceae species: CMA+ /DAPI-, CMA- /DAPI+ and CMA0/DAPI+ bands, varying in the pattern of distribution, number and size of heterochromatic bands. CMA+/DAPI- bands were observed in all species analyzed, and probably is the type of heterochromatin associated with RONs.The species analyzed ranged from 2n = 18 para Chamaecostus subsessilis (Costaceae) to 2n = 76 in Dichorisandra hexandra (Commelinaceae). Polyploidy and dysploidy are the main mechanisms of karyotype evolution involved in the diversification in Apocynaceae, with the diploid species being most frequently found in inselbergues. Among the analyzed genera there is no clear relationship between the number of CMA+ bands and the level of ploidy. Of the 28 species occurring in inselbergues in northeastern Brazil, only seven are polyploid, occurring especially in the Apocynaceae, Commelinaceae, Euphorbiaceae and Melastomataceae families. In addition, the preferential occurrence of polyploid cytotypes on inselbergues was not observed for these species, refuting the hypothesis that poliploids present greater tolerance to more extreme edaphoclimatic conditions. Keywords: Outcrops, Angiosperms, Cytogenetics, CMA/DAPI, Ploidy LISTA DE TABELAS Capítulo I Tabela 1. Espécies da família Apocynaceae analisadas por meio de dupla coloração com fluorocromos. Subfamília, gênero e espécie, local de coleta, coletor e número, fórmula cariotípica (FC), contagens cromossômicas prévias (CP),
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