Resumo: Vassoura-de-bruxa e podridão de Moniliophthora, causadas pelos fungos Crinipellis perniciosa e Moniliophthora roreri, respectivamente, são as doenças de maior impacto econômico da cultura do cacau e estão presentes na maioria dos países produtores do Continente Americano. Evidências biológicas e moleculares comprovam que estes fitopatógenos estão intimamente relacionados. O uso de resistência genética através de dones resistentes de cacaueiro, é a medida mais eficiente no controle destas doenças. O conhecimento sobre as populações destes fungos é importante na geração de informações para o programa de melhoramento genético do cacau visando resistência. Marcadores moleculares RAPO e SSR foram usados para analisar a estrutura genética de populações destes fitopatógenos. No geral, as populações do Brasil, Equador, Peru e Trinidad agruparam-se de acordo com o país de origem, apresentando maior variabilidade dentro e não entre países, com presença de subpopulações. A população do Brasil apresentou maior diversidade genotípica em comparação com as demais. A transferibilidade de pares de primers SSR de C. perniciosa para M. roreri foi satisfatório. Populações de M. roreri do Equador e Peru apresentaram alta diferenciação genética interpopulacional, sendo que a do Peru apresentou maior variabilidade. / Abstract: Witches' broom and fresty pod hot, caused by Crinipellis perniciosa and Moniliophthora roreri, are the most important disease of cacao in the American Continent. Biological and molecular data have shown that these pathogen are closely related. Resistance is the most efficient method to control these diseases. Therefore, information about the population structure of these cacao pathogen are important to support the breeding programo Molecular markers such as RAPD and SSR were used to analyzed the genetic structure of C. perniciosa and M. roreri frem the American Continent. Populations of C. perniciosa clustered according to their country of origin, with more variability within than between countries, revealing the presence of subpopulations. C. perniciosa Brazilian populations presented higher genotypic diversity than C. perniciosa from other countries. The transferability of C. perniciosa-SSR to M. roreri was positive. On the contrary, high interpopulation variability was observed between Ecuador and Peru, being M. roreri from Peru much more diverse than Ecuador. / Orientador: Carlos Ruggiero / Coorientador: Karina Peres Gramacho / Banca: João Carlos de Oliveira / Banca: Antonio de Goes / Banca: Maria Lúcia Carneiro Vieira / Banca: João Alexio Scarpare Filho / Doutor

In Canada, colorectal cancer is the second leading cause of death from cancer in men and the third leading cause of death from cancer in women. Several factors contribute to the development of cancer. Genetic predisposition, diet, and lifestyle habits are some of the major factors for colorectal cancer development. In the diet related factors, epidemiological studies suggest that consumption of whole grains rich in dietary fiber are associated with low incidence of human colon cancer. Recent studies have also shown that, in addition to dietary fiber, the type of dietary fiber and other compounds such as phenolic acids present in cereal grain bran may also have a role to play in colon cancer prevention. In a recent study, eleven major phenolic acids which differed in anti-oxidant activity were identified in wheat bran from wheat varieties belonging to six different market classes. The main objective of this study was to develop an in vitro cell culture based assay system to study the effect of phenolic acids on colon cancer development. Another objective was to study the effect of phenolic acids on selected molecular markers associated with cell proliferation, apoptosis and inflammation. Two well established colon carcinoma cell lines HT-29 and HCT 116 were treated with varying concentrations of fourteen phenolic acids to study their effect on cell survival and proliferation. In addition, immunohistochemical assays were performed on treated cells for two cell proliferation markers (Cyclin D1 and Ki67), an apoptosis marker (Bax) and three inflammatory markers (Beta-catenin, COX-2 and iNOS). Treatment of phenolic acids inhibited the growth of both the cell lines, however the effects varied with phenolic acid and cell line used in the assay. As determined by IC50, the growth of HCT 116 cells was inhibited the most by caffeic, ellagic, and gallic acids with IC50 of 0.22 mM, 0.17 mM, and 0.15 mM, respectively. On the other hand, caffeic, chlorogenic, and gallic acids are most effective in preventing the growth of HT-29 cells, with IC50 at 0.06 mM, 0.28 mM, and 0.30 mM, respectively. Immunohistochemical and Western Blotting studies revealed that phenolic acids differentially affected markers for cell proliferation, apoptosis and cell inflammation. In most of the cases, phenolic acid treatments up-regulated the pro-apoptosis marker Bax, while it down-regulated cell proliferation marker Cyclin D1. The results clearly show that a cell culture based assay can be used to study the effect of phenolic acids or other chemical constituents isolated from plants to study their effect on colon cancer cell lines. Statistical analysis revealed that only in very limited cases, results of molecular markers correlated to cell growth and proliferation. Therefore, to draw firm conclusions, more detailed and extensive studied need to be completed using different phenolic acids, the two cell lines and more replications. However, this study has developed the necessary protocols and provided some indicative results such as most of the phenolic acids induced pro-apoptosis pathway in both the colon cancer lines. Future studies with extracted phenolic acids from wheat bran using the cell culture system optimized in this study can be used to define the role of different wheat varieties in colon cancer prevention.

Population based association studies in crops that were established by domestication and early breeding can be a valuable basis for the identification of QTLs. A case control design in a population is an ideal way to identify maximum candidate sites contributing to a complex polygenic trait such as drought. In the current study, marker loci associated with drought related QTLs were identified in sugarcane (Saccharum spp), one of the most complex crop genomes, with its polyploid nature (>8), chromosome number (>100) and interspecific origin. The objectives of this investigation were: 1) development of genic markers, which can be used for marker-assisted selection of drought tolerant genotypes of sugarcane. 2) genotypic characterization of sugarcane population at drought related loci using EST-SSR markers. Using 55 microsatellite markers, 56 polymorphisms were scored among 80 modern sugarcane genotypes. Homogeneity of the population was confirmed by determining the distribution of allele frequencies obtained by random genomic microsatellite markers. This analysis was conducted in the STRUCTURE program and the population was divided in 3 subgroups based on the allelic distribution. Phenotypic data to evaluate drought tolerance among the genotypes was collected by measuring chlorophyll content, chlorophyll fluorescence, leaf temperature and leaf relative water content. A generalized linear model in SPSS was used to find association between marker loci and phenotypic data. Markers with significant association (P 0.001 level) with the trait were subjected to linear regression to screen the spurious associations. Based on the results, 21 EST-SSR markers and 11 TRAP markers related to drought-defining physiological parameters were considered as genuine associations in this study. Fifty-six polymorphisms produced by 13 EST-SSR primers were used to produce genetic similarity matrix for 80 genotypes. Dendrogram prepared from this genetic similarity matrix will be useful in selecting parents carrying diversity at drought specific loci.

The objective of this thesis was to investigate the genomic changes that have occurred due to the effects of long-term artificial selection applied by soybean breeders. A total of 42 cultivars from six different breeding programs, comprising the multi-generational pedigree of OAC Bayfield were genotyped with molecular markers and chromosomal inheritance was tracked throughout the pedigree. The graphical genotype profile of the 20 chromosomes revealed substantial allelic structure that has been built up in certain chromosomes, in the form of specific linkage blocks, which have been conservatively inherited. A selective sweep analysis using microsatellite markers was performed using the members of OAC Bayfield’s pedigree to identify genomic regions that have retained a molecular selective signature through OAC Bayfield in the varieties derived from it. Overall, there was a high level of agreement between the identified quantitative trait loci (QTL) and the phenotypic traits that would have been expected to be under breeders’ selection.

Soybean (Glycine max) is an important crop in the world in terms of total production and usage. It is also among the least diverse species. The main objectives of the present study were 1) to determine differences between ISSR and RAPD marker systems in detecting genetic variation in soybeans and 2) to identify and characterize accession- diagnostic molecular markers in G. max accessions. Genomic DNAs from 108 G. max accessions from 11 different gene pools were analyzed using several ISSR and RAPD primers. The levels of polymorphic loci detected with the two marker systems were in general moderate and similar.. Overall, 82% of genetic distance values were above 0.40 based on ISSR analysis. However, RAPD data revealed that the accessions from different countries are closely related with 64% genetic distance values below 0.40. The dendrograms constructed with ISSR data revealed that the South Korean accessions formed an out-group while the RAPD analysis showed that accessions from Sweden were separate from the other 10 gene pools. One variety-diagnostic marker generated with ISSR 5 primer was identified in the accession Kao Chien Tao from China. This marker was cloned, and sequenced. Although RAPD and ISSR marker systems detected similar levels of genetic variability, they target different regions of the soybean genome, resulting in different clustering of the 11 gene pools indicating different genetic relatedness among them. This finding demonstrates the usefulness of both marker systems in assessing diversity and relatedness among Glycine max gene pools.

MELO, José Glauber Moreira. Análise molecular e proteômica de Lasiodiplodia theobromae associado a fruteiras tropicais. 2014. 88 f. Tese (Doutorado em agronomia)- Universidade Federal do Ceará, Fortaleza-CE, 2014. / Submitted by Elineudson Ribeiro (elineudsonr@gmail.com) on 2016-08-29T20:06:07Z No. of bitstreams: 1 2014_tese_jgmmelo.pdf: 2467689 bytes, checksum: e9a0fc6142e5ef3db09ac0252d32f4e7 (MD5) / Approved for entry into archive by Jairo Viana (jairo@ufc.br) on 2016-08-31T23:39:17Z (GMT) No. of bitstreams: 1 2014_tese_jgmmelo.pdf: 2467689 bytes, checksum: e9a0fc6142e5ef3db09ac0252d32f4e7 (MD5) / Made available in DSpace on 2016-08-31T23:39:17Z (GMT). No. of bitstreams: 1 2014_tese_jgmmelo.pdf: 2467689 bytes, checksum: e9a0fc6142e5ef3db09ac0252d32f4e7 (MD5) Previous issue date: 2014 / Lasiodiplodia theobromae is a plant pathogenic fungus responsible for many diseases in various plants, is a fungus typically tropical and subtropical regions. The fungus attacks many tropical plants, among them, including mango, Spondias sp., coconut, cashew, and many others. His control is basically genetic performed by planting resistant clones, however to obtain it becomes necessary to know the pathogen characteristics. The information available on the genetic variability of L. theobromae is restricted to ensure success in any breeding program for resistance to this pathogen. Taking into account that proteins are functional products of genes, it is important to know them, to improve the understanding of the mode of action of pathogens, with this understanding, useful as a strategy to be used in plant breeding seeking genetic resistance. The objective of this study were to conduct a genetic study molecular in a population of L. theobromae and a differential proteomic analysis of the fungus among isolates, more and less aggressive, to identify proteins responsible for this aggressivity. A population consisting of 105 strains was used for molecular characterization, extracting the DNA from mycelia grown in liquid medium. Each sample was subjected to polymerase chain reaction (PCR) with 15 pairs of primers specific for the species, and a primer pair of the ITS region and other EF-1α region. The amplified products were visualisados agarose gel, stained with ethidium bromide and spreadsheet data in binary tabulated and analyzed by unbalanced grouping method based on the arithmetic mean (UPGMA) using the MVSP program. Genetic similarities were estimated by Jaccard’s coefficient. The results indicated a high genetic variability of the studied population. Since the proteomic study was conducted to assess qualitative differences, that is, the presence/absence of a specific protein in relation to the opposite group. To this end, we used two isolates of the same fungus, differing as their aggressiveness, in which one was highly aggressive, whereas the other had a low aggressiveness when inoculated seedlings of cashew. When the electrophoretic profile was analyzed, 96 were detected differentially expressed spots. By LC-ESI-Q-TOF MS / MS, 84 proteins were identified having the most diverse cellular functions. With this approach it was possible preliminary characterization of the protein profile of this fungus to give some evidence of the mechanisms involved in their aggressiveness. This is the first study seeking to know the proteins responsible for the aggressiveness L. theobromae. / Lasiodiplodia theobromae é um fungo fitopatogênico responsável por inúmeras doenças em variadas plantas, sendo um fungo tipicamente de regiões tropicais e subtropicais. O fungo atacadiversas plantas tropicais, dentre elas destacam-se a mangueira, as Spondiassp., o coqueiro, o cajueiro, entre tantas outras. Seu controle é basicamente genético realizado com o plantio de clones resistentes, entretanto, para sua obtenção torna-se necessário o conhecimento das características do patógeno. As informações disponíveissobre a variabilidade genética de L. theobromaesão insuficientes para assegurar o sucesso em qualquer programa de melhoramento genético visando à resistência a este patógeno. Levando-se em conta que as proteínas são produtos funcionais dos genes, torna-se importante conhecê-las, visando um melhor entendimento do modo de ação dos patógenos, sendo este entendimento, útil como estratégia a ser utilizada no melhoramento vegetal buscando a resistência genética. Assim, o objetivo desse estudo foi realizar um estudo genético molecular em uma população de L. theobromae e uma análise proteômica diferencial do fungo entre isolados, mais e menos agressivos, visando identificar proteínas responsáveis por essa agressividade.Uma população composta de 105 isolados foi usada na caracterização molecular, extraindo-se o DNA a partir do micélio do fungo crescido em meio líquido. Cada amostra foi submetida à reação em cadeia da polimerase (PCR) com 15 pares de primers específicos para essa espécie, além de um par de primer da região ITS e outro da região EF-1α. Os produtos amplificados foram visualisados em gel de agarose, corados com brometo de etídio e os dados tabulados em planilha binária e foram analisados pelo método de agrupamento não balanceado baseado na média aritmética (UPGMA) utilizando o programa MVSP. As similaridades genéticas foram estimadas pelo coeficiente de Jaccard. Os resultados indicaram uma grande variabilidade genética da população avaliada. Já o estudo proteômico foi realizado visando avaliar diferenças qualitativas, ou seja, a presença/ausência de uma determinada proteína, em relação ao grupo antagônico. Para tal, utilizaram-se dois isolados do mesmo fungo, diferenciando-se quanto a sua agressividade, em que um era altamente agressivo, enquanto o outro apresentava uma baixa agressividade quando inoculados em mudas de cajueiro. Quando o perfil eletroforético foi analisado, foram evidenciados 96 spots diferencialmente expressos. Através da LC-ESI-Q-TOF MS/MS, foram identificadas 84 proteínasapresentando diversas funções celulares.Com essa abordagem foi possível a caracterização preliminar do perfil proteico deste fungo, obtendo-se alguns indícios dos mecanismos envolvidos na sua agressividade. Este é o primeiro estudo buscando conhecer as proteínas responsáveis pela agressividade de L. theobromae.

Current breeding of oilseed rape is focused on breeding of F1 hybrids and the male sterility and self-incompatibility could play the significant role in hybrid breeding programmes. Plant breeders also more widely utilize molecular genetic techniques for selection of desirable plants/genotypes. Molecular methods are faster, more reliable and more specific than conventional ones, which are based mainly on morphological descriptors. The aim of my thesis was to optimize PCR analysis methods and to develop specific molecular markers for target plant selection in oilseed rape hybrid breeding programmes. The new marker for detection of fertility restoration gene (Rf) in CMS Ogu-INRA plants was tested. New primer pair RsPPRF2/RsPPRR2 was designed in the coding region of PPR-B protein, which participates in the restoration of fertility in CMS Ogu-INRA. Also newly designed primers BrSLGIIF/BrSLGIIR were tested in SI plants. The optimal annealing temperaturse of these primers was 58 °C. But amplification in some SC plants was also observed. The optimization of the PCR reaction was performed for all designed primers. The set of F1 SI hybrids created by crossing of two lines with different S II haplotypes was tested by using of the PCR-RFLP technique for detection of polymorphism in amplified fragments.

Made available in DSpace on 2014-06-11T19:26:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-26Bitstream added on 2014-06-13T19:33:13Z : No. of bitstreams: 1 prado_fd_me_botib.pdf: 676910 bytes, checksum: 6d6c9b4d7738938a01e94ca150d37e4a (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A hibridação artificial interespecífica de peixes é utilizada em diversos estabelecimentos voltados à piscicultura no país, com a finalidade de produzir indivíduos mais vantajosos e favoráveis para o cultivo. Porém, devido principalmente às dificuldades encontradas na identificação dos híbridos, esta prática pode determinar o surgimento de sérios problemas como a contaminação genética dos estoques de cultivo, a comercialização de produtos híbridos como espécies puras, a introdução de espécies exóticas e escapes de produtos de piscicultura para o ambiente natural e até eventos de introgressão genética e extinção das espécies nativas. Neste contexto, o presente trabalho teve por objetivo analisar geneticamente exemplares das linhagens parentais de Pseudoplatystoma corruscans (pintado) e Pseudoplatystoma reticulatum (cachara) e seus híbridos interespecíficos pintachara (macho de pintado x fêmea de cachara) e cachapinta (fêmea de cachara x macho de pintado), provenientes dos estoques de cultivo do CEPTAlICMBio, Pirassununga, SP, a fim de identificar e estabelecer marcadores genéticos para possibilitar sua identificação e diferenciação. Também foram analisadas geneticamente amostras de P. corruscans e P. reticulatum coletadas no Rio Paraguai, MS (bacia do Paraguai) e de P. corruscans capturados do rio Mogi- Guaçu, SP (bacia do Alto Paraná), a fim de identificar a possível ocorrência de híbridos entre estas espécies na natureza. As análises citogenéticas revelaram um número diploide de 2n=56 cromossomos para ambas as espécies parentais, com cariótipos caracterizados por cromossomos dos tipos 20m+12am+12st+12a e número fundamental (NF) igual a 100, indicando uma fórmula cariotípica conservada entre estas espécies. A análise dos padrões de heterocromatina pelo bandamento C revelou blocos heterocromáticos localizados nas porções... / Artificial interspecific hybridization of fish is used in various establishments linked to fish farming in the country, in order to produce individuais more advantageous and favorable for cultivation. However, due mainly to difficulties in the hybrid products identification, this practice may determine the onset of serious problems such as genetic contamination of the breeder stocks, marketing of hybrids as pure species, introduction of exotic species and escapes of farmed products to the natural environment and sometimes leading to events of genetic introgression and extinction of native species. In such context, the present study aimed to analyze genetically copies of the parental lines of Pseudoplatystoma corruscans (pintado) and Pseudoplatystoma reticulatum (cachara) and their interspecific hybrids pintachara (female of pintado x male of cachara) and cachapinta (female of cachara x male of pintado), from the stocks manteined in CEPTAlICMBio, Pirassununga, SP, in order to characterize and establish genetic markers to allow their identification and differentiation. Samples of P. corruscans and P. reticulatum collected in the Paraguay river, MS (Paraguay basin) and P. corruscans captured in Mogi-Guaçu river, SP (Alto Paraná basin), also were genetically analyzed in order to identify the possible occurrence of hybrids between these species in nature. The cytogenetic analysis revealed a diploid number of 2n = 56 chromosomes for both parental species with karyotypes characterized by the formula 20m+12am+12st+12a and fundamental number (NF) of 100, indicating a karyotypic formula conserved among these species. The analysis of the heterochromatin C-banding patterns revealed heterochromatic blocks located in the pericentromeric and terminal portions of some chromosomes, the NOR was sim pie and located in one chromosome pai r and 5S ribosomal genes located on two different subtelocentric... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:26:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2003-11-28Bitstream added on 2014-06-13T18:29:33Z : No. of bitstreams: 1 ganga_rmd_me_jabo.pdf: 819507 bytes, checksum: 5ba45956cafb05a742d55332f642a8b2 (MD5) / Os maracujazeiros pertencem à família Passifloraceae e ao gênero Passiflora, reunindo mais de 500 espécies distribuídas pelos trópicos, principalmente no Brasil, centro de origem de pelo menos um terço das espécies, o que determina uma grande variabilidade genética. Como maior produtor mundial da fruta, o Brasil tem cerca de 35 mil hectares de área colhida e produção superior a 317 mil toneladas por ano, no qual Bahia, São Paulo e Sergipe respondem por mais de 50% da produção no País. A avaliação da variabilidade presente é indispensável aos trabalhos de melhoramento genético, cuja caracterização molecular pode fornecer a base para estudos de diversidade, pautando-se na composição genética sem influência do ambiente. Marcadores moleculares fAFLP foram utilizados para estimar a diversidade genética entre 36 acessos de maracujá amarelo (Passiflora edulis f. flavicarpa Deg.) coletados em 18 estados do Brasil. Os resultados obtidos permitiram concluir que os marcadores fAFLP se mostraram consistentes na verificação da variabilidade genética, detectando e quantificando a ampla divergência genética entre os 36 acessos de Passiflora edulis f. flavicarpa Deg. analisados, bem como a não formação de estruturação geográfica. Tais resultados podem auxiliar na definição de estratégias mais eficientes a serem utilizadas em programas de melhoramento genético de maracujazeiro amarelo. / Passion fruit trees belong to the Passionfloraceae family and to the Passiflora genus, gathering more than 500 species distributed over the tropics, mainly in Brazil, source of at least one third of the species, what determines a great genetic variability. As the world's biggest producing country, Brazil has around 35 thousand hectares of harvest area, and a production superior to 317 thousand tons per year from which Bahia, São Paulo and Sergipe are responsible for more than 50%. The assessment of the variability is essential to genetic breeding works, whose molecular characterization can provide us with the basis for studies on diversity, taking into account the genetic composition without environmental influence. fAFLP molecular markers were utilized to estimate genetic diversity among 36 yellow passion fruit accessions (Passiflora edulis f. flavicarpa Deg.) colleted in 18 Brazilian states. The obtained results led to the conclusion that the fAFLP markers were consistent regarding to the evaluation of genetic variability, detecting and quantifying the ample genetic divergence among the 36 analyzed accessions, as well as to the no geographic structural formation. Such results can be useful to the definition of more efficient strategies to be applied in genetic breeding programs of yellow passion fruit tree.

Made available in DSpace on 2014-06-11T19:32:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-12-09Bitstream added on 2014-06-13T19:43:14Z : No. of bitstreams: 1 mendonca_pc_dr_botfca.pdf: 892807 bytes, checksum: c81f229a150e1ff79516536ed6fe7f98 (MD5) / A espécie Stryphnodendron adstringens (Leguminosae) é conhecida popularmente como barbatimão e o extrato das cascas é utilizado como cicatrizante. O objetivo deste trabalho foi estudar a variabilidade genética da espécie utilizando o marcador molecular de polimorfismo de comprimento amplificado (AFLP) e testar a transferência de microssatélites de Anadenanthera colubrina, Hymenaea courbaril e Copaifera langsdorffii. Foram coletados acessos localizados nos municípios de Cristalina, São João D’Aliança, Campo Alegre e Caldas Novas (GO); Delfinópolis, Luislândia, Lagoa Formosa, Sacramento e Araxá (MG) e Paranapanema, Cristais Paulista e Botucatu (SP). O DNA genômico foi extraído de folhas e as análises de polimorfismo seguiram as etapas de digestão, ligação, pré-amplificação e amplificação. Os produtos AFLP foram separados em gel desnaturante de poliacrilamida 6% com tampão TBE 1X. A eletroforese foi realizada em voltagem constante de 80W em temperatura máxima de 50ºC por 4 horas. O gel foi corado com solução de nitrato de prata e revelado em carbonato de sódio. Na análise por marcador AFLP foram produzidas 237 bandas polimórficas. A variabilidade dentro das populações foi maior (70,93%) que entre as populações (29,06%) com um valor de Fst 0.2906 indicando alta estruturação populacional. A população de Luislândia apresentou maior porcentagem de loci polimórficos (87,35), seguida da população de Cristalina (45,85). A menor variabilidade foi encontrada em Caldas Novas (22,92) e as demais ficaram na média (34,3). O Método da Média Aritmética não Ponderada (UPGMA) reuniu as populações em três grupos. Quanto aos testes de transferência de microssatélites, dos 20 iniciadores de A. colubrina testados, dez apresentaram resultados de transferência, porém somente um... / Stryphnodendron adstringens a Leguminosae species is popularly known as barbatimão and the extract of its barks is widely used as healing agent. The genetic variation of 12 populations of S. adstringens was determined in this study by using Amplified Fragment Length Polymorphism (AFLP) molecular markers transference of microsatellites of Anadenanthera colubrina, Hymenaea courbaril and Copaifera langsdorffii. Accessions were collected in the cities of Cristalina, São João D’Aliança, Campo Alegre and Caldas Novas (GO), Delfinópolis, Luislandia, Lagoa Formosa, Sacramento and Araxá (MG) and in Paranapanema, Cristais Paulista and Botucatu (SP). The genomic DNA was extracted from the leaves and the polymorphism analysis followed multiple steps including DNA digestion, ligation, pre-amplification and amplification. Amplification products were analyzed on denaturing polyacrylamide sequencing gel and running electrophoretic steps at 80 W with maximum temperature at 50ºC for 4 h. The gel was stained with silver nitrate solution and developed in sodium carbonate. The AFLP analysis conducted with three primer combinations using the EcoRI and MseI restriction enzymes generated 237 polymorphic bands. The AFLP binary data were used to determine allele frequencies. Population structure was evaluated performing analysis of molecular variance (AMOVA) which allowed the estimation of the total genetic variance among and inside populations. A descriptive analysis of the total variability was obtained by calculating the percentage of polymorphic loci. Genetic variance 4 within populations was higher (70,93%) compared to the differentiation estimated among populations (29,06%). The fixation index (Fst) was 0.2906 indicating highly significant population structuring. The population from Cristalina... (Complete abstract click electronic access below)