Anther culture experiments were conducted to construct a monoploid family. The donor plants used were hybrids between high leptine producing selections of Solanum chacoense Bitt. and anther culture responsive selections of Solanum phureja Juz. et Buk. Several steps of the anther culture process were studied. The results indicated that genotype remains the main factor affecting anther culture response. Growing anther-donor plants in higher greenhouse temperatures (30 degrees C day/20 degrees C night) increased the number of embryos per anther by 40 percent. A heat shock given to anthers in culture for 12h at 35 degrees C was also found to be beneficial resulting in an increase of the anther culture response by 40 percent. However the effect of the high temperature shock resulted in lower regeneration rates. In all experiments a highly significant "date" effect was observed with one or two days differing from the others by showing higher response rates in all hybrids tested. The majority of the regenerated plants was diploid, probably resulting from unreduced gametes. Simple sequence repeat analysis with eight polymorphic primer pairs was used successfully to identify the homozygous diploid plants that were added to the monoploids. In total 34 monoploid plants and 14 homozugous diploids were obtained. The degree of heterozygosity revealed by SSR analysis indicated that the diploid plants originated from unreduced gametes formed by first division restitution (FDR) mechanism. The SSR marker data were used to map the genes with respect to the centromeres by half tetrad analysis. SSR-containing sequences from the public databases, as well as sequences obtained from a genomic library enriched for SSRs, were used to generate 48 primer pairs. Only 12 of them were found to be polymorphic in the monoploid family. Ten primer pairs did not amplify any specific fragment. The monoploid population showed distorted segregation at four of the polymorphic loci, showing overrepresentation of the chacoense alleles in three of them. One of the loci showing distorted segregation (STSTP, amplified by primer pair RV 11+12) is most probably linked to lethal alleles, whereas another one (ST13ST, amplified by primer pair RV 21+22) could be linked to genes affecting anther-culture response. The location of the SSR loci on the potato chromosomes is not known except for one (waxy, primer pair 3+4), but statistical analysis on the segregation data obtained from 70 heterozygous anther-derived diploids showed no linkage between them. The SSR primer pairs developed in this study might be useful in studying genetic relationships among cultivars and accessions in breeding programs. Randomly amplified polymorphic DNA (RAPD) analysis was used in association with bulked segregant analysis to detect linkage with genes controlling leptine biosynthesis. With all the limitations imposed by the population size and contamination from foreign pollen, a band amplified by primer OPA-16 could differentiate the bulks contrasting for leptine content. It is possible that this band is linked to genes suppressing leptine biosynthesis, since it appears only in the plants that do not synthesize leptines. Further investigation with larger populations is needed to confirm this possibility. / Ph. D.

Mate competition is a prominent component of sexual selection theory. Typically, males attempt to mate with the most females possible and females attempt to mate with the highest quality males possible. In the polygynous female-defense mating system of Anolis carolinensis, males compete directly for females through territorial behavior. Inter-male competition is intense due to an average polygyny ratio of 1 male to 3 females despite a 1:1 adult sex ratio. Through high levels of territorial behavior (e.g., 100 displays/h, 27 m patrol distances/h, 70% of day in defense-related activities), males attempt to exclude other males from resident females who, in turn, both store sperm and ovulate a single-egg clutch at weekly intervals over a 4-month breeding season. Paternity of hatchlings in 16 naturally occurring breeding groups was analyzed to determine the extent to which the territorial resident male was able to prevent other males from fathering offspring of his resident females. Lizards residing in or neighboring a resident male's territory were collected and RAPD-PCR was used to determine the paternity of hatchlings. Of the 48 hatchlings from 26 females, resident territorial males fathered 52% of hatchlings; 15% were fathered by a male whose territory bordered that of the resident male and 21% were fathered by a smaller male living covertly within the resident male's territory. Paternity for the remaining 12% of hatchlings belonged to an unsampled male. Given that females mated with multiple males, laboratory-based controlled matings were conducted where females were sequentially paired with two males and RAPD-PCR was used to analyze which of the two males fathered the subsequent hatchlings to determine the mechanism of sperm precedence. Regardless of mating order, only one male of the pair fertilized the eggs. Male A. carolinensis have reproductive strategies present in addition to defending resident females and female A. carolinensis have options in addition to simply mating with the resident male. While sperm precedence is present in this species, it is not based on mating order, but may involve both the number of sperm deposited in the female's tract as well as the quality of those sperm. / Ph. D.

Colchicine, an antimicrotubule agent, has enhanced the androgenic response of cruciferous and graminaceous species. Our objectives were to determine if colchicine treatments enhanced androgenic embryo production in diploid potato, and if colchicine treatments during anther culture affected the ploidy of anther-derived plants. An additional study, analysis of genetic identity among regenerated anther-derived plants based on RAPD markers, was also conducted. Anther culture of a hybrid between <i>Solanum chacoense</i> and <i>S. phureja</i> (clone CP2) was conducted with five colchicine treatments (0, 25, 50, 100, and 200 mg/L) for 24 h. Anthers containing late uninucleate pollen were cultured on modified liquid LS medium. A mean of 0.25 embryos per anther was obtained; however, there was no significant difference among the colchicine treatments, which ranged from a mean of 0.17 to 0.28 embryos per anther for 200 mg/L colchicine and control, respectively. In total 56 plants were regenerated of which 11% were monoploid. The second experiment using the same colchicine treatments was conducted with 54 hybrids derived by crossing six anther-derived doubled monoploids with three heterozygous pollinators. A mean of 1.78 embryos per anther was obtained; again, there was no significant difference among the colchicine treatments, which ranged from 1.27 to 2.75 for 50 mg/L colchicine and control, respectively. In total 679 plants were regenerated. The 312 plantlets for which ploidy was determined, 75% were monoploid. The third experiment was conducted to study various durations (0, 90 sec vacuum infiltration, 24, 48, and 72 h) of high colchicine treatment (200 mg/L) applied to <i>S. phureja</i> family DM 13-14 202 x ID 5. There was nearly significant difference among treatments (α = 0.05). Mean embryos per anther ranged from 0.96 to 1.90 for 48 h and 90 sec vacuum infiltration, respectively. In an additional study, 26 regenerated anther-derived monoploid plants, from the DM 13-14 202 x ID 5 family, from the second experiment, were genetically characterized based on RAPD markers. Forty-three loci were scored from 13 primers for groups of monoploids derived from the same flasks of anther culture. From one flask, two pairs of monoploids were found to be genetically identical. From a second flask 6 of 7 were genetically identical, and from a third 3 of 7 were genetically identical. The presence of genetically identical individuals within three monoploid populations indicates the prevalence of secondary embryogenesis during anther culture such that a single embryogenic microspore can generate many anther-derived plants. / Master of Science

Made available in DSpace on 2016-08-10T10:55:06Z (GMT). No. of bitstreams: 1 Nicole Cristina Lopes Dutra.pdf: 1587085 bytes, checksum: 7c8f39f69da6456b027da52bf00a0794 (MD5) Previous issue date: 2006-11-29 / Studies that concern Cerrado (Brazilian s Savannah) fauna are important as we consider the elevated rate of destruction of its landscapes in the last decades. Bothrops moojeni is one of the species of vipers that occur on the Cerrado area and its venom has shown great applicability in health. In the last years, molecular markers have helped in researches that aim to assess the genetic variability. In the present work, we perform the characterization of 5 populations of Caiçaca (Bothrops moojeni) using RAPD (Random Amplified Polimorphyc DNA) markers and also used a scienciometric approach to highlight the importance of this species in health in the site Institute for Scientific Information (ISI), using the method General Search with the key words: serpente , Bothrops and Bothrops moojeni . The obtained Data were used to estimate the magnitude and distribution of variability within and among the groups using AMOVA. Aiming to analyze the patterns of spatial variation patterns, Pearson relation coefficient (r) was estimated between the matrixes of genetic distances and geographic among the populations. With the 5 primers were obtained 59 loci, from which 81 porcent (48) were polymorphic, ranging between 35% and 69% in the groups. The value of f ST obtained with AMOVA was equal to 0,13 (P smaller 0,0001 - 10000 permutations). Thus, it was possible to observe that Caiçaca populations studied presented a low level of genetic variability (He = 0,19) and most of the genetic variation is within the populations, what might indicate that is already occurring a reduction of the rate of gene flow between the populations. Considering the geographic area studied (lager distance was equal to 851,83 km), populations that are geographically near presented a high level of genetic diversity and the genetic variability is not spatially structured. The populations of Bothrops moojeni studied presented low levels of Genetic diversity for the RAPD loci evaluated. Genetic variability is not spatially structured. Concerning scienciometry analyses, the following key-words: serpente , Bothrops and Bothrops moojeni most of the studies concerning zoology (51 porcent), Pharmacology and Pharmacy (32 porcent) and Biochemistry and Molecular Biology (23 porcent), there was no paper published till the present date, that aimed to evaluate genetic variability in this species, as well as in the genus Bothrops. We recommend a wide-ranging study to evaluate genetic variability in this species due to its importance to conservation of the ecosystem from which it belong as well due to the great interest in health. / Estudos sobre a fauna do Cerrado são importantes considerando a alta taxa de destruição das suas paisagens naturais durante as últimas décadas. Estudos com marcadores moleculares se fazem importantes, pois auxiliam nas pesquisas que pretendem acessar a variabilidade genética para fins de conservação. Entre as espécies do Cerrado, Bothrops moojeni é uma serpente que vem demonstrando grande aplicabilidade na saúde. Neste sentido, o objetivo deste trabalho foi caracterizar a variabilidade genética de cinco populações de Caiçaca (Bothrops moojeni Hoge, 1966) a partir de marcadores RAPD (Random Amplified Polimorphyc DNA), bem como realizar um levantamento cienciométrico da importância da espécie para a área da saúde no sítio Institute for Scientific Information (ISI), utilizando o método GeneralSearch com a palavras chaves: serpentes, Bothrops e Bothrops moojeni. Os dados gerados com marcadores RAPD foram utilizados para avaliar a magnitude e a distribuição da variabilidade entre e dentro das populações por meio da Análise de Variância Molecular (AMOVA). A fim de se avaliar os padrões de variação espacial, foi estimado o coeficiente de correlação de Pearson (r) entre a matriz de distâncias genéticas e geográficas entre as populações. Com base nos cinco primers foram obtidos 59 locos, dos quais 81 por cento (48) foram polimórficos, variando entre 35 por cento e 69 por cento nas populações. O valor do Fí ST obtido pela AMOVA foi igual a 0,13 (P menor 0,0001 - 10000 permutações). Foi possível observar que as populações de Caiçaca estudadas apresentam baixos níveis de variabilidade genética (He = 0,19) para os locos RAPD estudados, sendo que a maior parte da variação encontra-se dentro das populações, o que poderia indicar que já está ocorrendo uma redução na taxa de fluxo gênico entre as populações. Considerando a escala geográfica em estudo (maior distância igual a 851,83 km), populações que estão próximas geograficamente apresentam uma considerável divergência genética e a variabilidade genética não se encontra estruturada espacialmente. No que diz respeito a cienciometria, nas palavras-chave serpente , Bothrops e Bothrops moojeni a maioria dos estudos foi sobre zoologia (51 por cento), Farmacologia e Farmácia (32 por cento) e Bioquímica e Biologia Molecular (23 por cento), não existindo nenhum trabalho publicado, até a presente data, que avalie a variabilidade genética desta espécie. Deve-se proceder com uma análise mais sistemática e de maior magnitude para avaliar a variabilidade genética desta espécie que além de sua importância para a conservação é uma espécie de muito interesse para a área de saúde.

Made available in DSpace on 2017-07-10T17:37:30Z (GMT). No. of bitstreams: 1 Vanessa Aparecida Antes.pdf: 571702 bytes, checksum: c629643a9f77bdb02f4dd1641cbecf68 (MD5) Previous issue date: 2008-08-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The aim of this work was to study the parasitism of Meloidogyne incognita on native plants from Western Paraná, Brazil, as well as to assess the genetic variability among different populations belonging to the race 3 of this nematode by RAPD technique. Natural infection was studied in thirty six native plant species, which were identified on the basis of the perineal pattern from mature females and esterase phenotype. Native species that showed no infestation on the root system were inoculated with 1.000 eggs and/or J2 of M. incognita. After 60 days, the inoculated plants were evaluated regarding the number of galls and the number of eggs/or J2 per root. The genetic variability from different single female populations of M. incognita race 3 was studied by RAPD technique, having been tested 10 primers. The native plants that were susceptible to M. incognita parasitism were Rabo-de-Bugio (Lonchocarpus muehlbergianus Hassl.), Ipê Roxo (Tabebuia impetiginosa Mart. Ex DC. Standl), Sanga D água (Croton urucurana Boill), Ipê Amarelo (Tabebuia ahrysotricha Mart ex DC. Standl.), Genipapo (Genipa americana L.), Ariticum Comum (Rollinia mucosa (Jacq.) Baill.) and Aroeira Vermelha (Schinus terebinthifolius Raddi.). On the other hand, M. javanica was found parasiting Café de Bugre (Cordia ecalyculata Vell.), Guatambu Vermelho (Aspidosperma subincanun Mart.) and Tarumã Branco (Cytrarexllum myruanthum Cham.). The DNA polymorphism showed that there was genetic variability among populations from a same race (3) of M. incognita, allowing the separation of them into five genetic groups, through reactions with the primers (O)AK20, A10, AQ12, AS08 and (OP)F01 / Objetivou-se com o presente trabalho estudar o parasitismo de Meloidogyne incognita em plantas nativas do oeste paranaense, bem como a variabilidade genética de populações de M. incognita raça 3 pela técnica RAPD. Trinta e seis espécies de plantas nativas foram analisadas com relação à infecção natural por Meloidogyne spp. A identificação de Meloidogyne foi feita com base na configuração perineal de fêmeas maduras e no fenótipo para a isoenzima esterase. Espécies nativas com ausência de galhas no sistema radicular foram inoculadas com 1.000 ovos e/ou J2 de uma população de campo de M. incognita. Após 60 dias da inoculação, as plantas foram avaliadas com relação ao número de galhas e número de ovos e/ou juvenis por raiz. A variabilidade genética de diferentes populações monoespecíficas de M. incognita raça 3 do noroeste paranaense foi estudada pela técnica RAPD com o teste de 10 primers. As plantas nativas hospedeiras de M. incognita foram Rabo-de-Bugio (Lonchocarpus muehlbergianus Hassl.), Ipê Roxo (Tabebuia impetiginosa Mart. Ex DC. Standl), Sanga D água (Croton urucurana Boill), Ipê Amarelo (Tabebuia ahrysotricha Mart ex DC. Standl.), Genipapo (Genipa americana L.), Ariticum Comum (Rollinia mucosa (Jacq.) Baill.) e Aroeira Vermelha (Schinus terebinthifolius Raddi.). Já M. javanica foi encontrado parasitando Café de Bugre (Cordia ecalyculata Vell.), Guatambu Vermelho (Aspidosperma subincanun Mart.) e Tarumã Branco (Cytrarexllum myruanthum Cham.). A análise do polimorfismo do DNA possibilitou a detecção de variabilidade genética para diferentes populações de uma mesma raça (3) de M. incognita, permitindo a separação das populações em 5 grupos genéticos, através de reações com os primers AK20, A10, AQ12, AS08 e F01

Made available in DSpace on 2016-08-12T19:15:42Z (GMT). No. of bitstreams: 1 AnaCAD_DISSERT.pdf: 992908 bytes, checksum: 8c8ed25d36c890fc913b87012539bb33 (MD5) Previous issue date: 2011-02-25 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The melon plant is distributed worldwide and is the kind that has the highest phenotypic variability of the genus, especially observed in its fruit. There is great variability which consists an important source of germplasm for genetic improvement programs. Therefore, the knowledge of the genetic variability of vegetable species and how it distributes itself, provides the rational and sustainable use of genetic resources. The objective of the present study was to do a morphological and molecular characterization of accessions of melon plant collected in brazilian northeastern. Were evaluated 40 accesses and three commercial cultivars in a trial conducted in randomized complete block with two repetitions in Mossoró-RN. The morphological characterization was performed through 17 descriptors, one of the seed, fourteen of fruit and two of the inflorescence. The molecular characterization was performed by RAPD markers using 18 primers and by microsatellite (SSR) with 15 primers synthesized for Cucumis melo. It was verified that the morphological markers, RAPD and SSR were satisfactory to allow the detection of polymorphism among the genotypes evaluated. The grouping methods of Tocher and the hierarchical partially agreed on morphological and molecular characterizations. The RAPD marker was more discriminating in relation to microsatellite by the fact of cluster the accesses in most groups. The germplasm bank of UFERSA has high genetic variability among the accessions. / O meloeiro está distribuído em todo o mundo, e é a espécie que possui a maior variabilidade fenotípica do gênero, observada principalmente nos seus frutos. Existe grande variabilidade que consiste em importante fonte de germoplasma para programas de melhoramento. Portanto, o conhecimento da variabilidade genética de espécies vegetais e como ela se distribui, proporciona o uso racional e sustentável dos recursos genéticos. O objetivo do presente trabalho foi realizar a caracterização morfológica e molecular de acessos de meloeiro coletados no Nordeste brasileiro. Foram avaliados 40 acessos e três cultivares comerciais em um experimento conduzido em blocos completos casualizados com duas repetições no município de Mossoró-RN. A caracterização morfológica foi feita através de 17 descritores, sendo um da semente, quatorze de frutos e dois da inflorescência. A caracterização molecular foi realizada pelos marcadores RAPD utilizando 18 primers e por microssatélites (SSR) com 15 primers sintetizados para Cucumis melo. Verificou-se que os marcadores morfológicos, RAPD e SSR foram satisfatórios em permitir a detecção de polimorfismo entre os genótipos avaliados. Os métodos de agrupamento de Tocher e o hierárquico concordaram parcialmente nas caracterizações morfológicas e moleculares. O marcador RAPD foi mais discriminante em relação ao microssatélite pelo fato de agrupar os acessos em mais grupos. O banco de germoplasma da UFERSA possui alta variabilidade genética entre os acessos.

Made available in DSpace on 2016-08-12T19:18:57Z (GMT). No. of bitstreams: 1 AnaCAD_DISSERT.pdf: 992908 bytes, checksum: 8c8ed25d36c890fc913b87012539bb33 (MD5) Previous issue date: 2011-02-25 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The melon plant is distributed worldwide and is the kind that has the highest phenotypic variability of the genus, especially observed in its fruit. There is great variability which consists an important source of germplasm for genetic improvement programs. Therefore, the knowledge of the genetic variability of vegetable species and how it distributes itself, provides the rational and sustainable use of genetic resources. The objective of the present study was to do a morphological and molecular characterization of accessions of melon plant collected in brazilian northeastern. Were evaluated 40 accesses and three commercial cultivars in a trial conducted in randomized complete block with two repetitions in Mossoró-RN. The morphological characterization was performed through 17 descriptors, one of the seed, fourteen of fruit and two of the inflorescence. The molecular characterization was performed by RAPD markers using 18 primers and by microsatellite (SSR) with 15 primers synthesized for Cucumis melo. It was verified that the morphological markers, RAPD and SSR were satisfactory to allow the detection of polymorphism among the genotypes evaluated. The grouping methods of Tocher and the hierarchical partially agreed on morphological and molecular characterizations. The RAPD marker was more discriminating in relation to microsatellite by the fact of cluster the accesses in most groups. The germplasm bank of UFERSA has high genetic variability among the accessions. / O meloeiro está distribuído em todo o mundo, e é a espécie que possui a maior variabilidade fenotípica do gênero, observada principalmente nos seus frutos. Existe grande variabilidade que consiste em importante fonte de germoplasma para programas de melhoramento. Portanto, o conhecimento da variabilidade genética de espécies vegetais e como ela se distribui, proporciona o uso racional e sustentável dos recursos genéticos. O objetivo do presente trabalho foi realizar a caracterização morfológica e molecular de acessos de meloeiro coletados no Nordeste brasileiro. Foram avaliados 40 acessos e três cultivares comerciais em um experimento conduzido em blocos completos casualizados com duas repetições no município de Mossoró-RN. A caracterização morfológica foi feita através de 17 descritores, sendo um da semente, quatorze de frutos e dois da inflorescência. A caracterização molecular foi realizada pelos marcadores RAPD utilizando 18 primers e por microssatélites (SSR) com 15 primers sintetizados para Cucumis melo. Verificou-se que os marcadores morfológicos, RAPD e SSR foram satisfatórios em permitir a detecção de polimorfismo entre os genótipos avaliados. Os métodos de agrupamento de Tocher e o hierárquico concordaram parcialmente nas caracterizações morfológicas e moleculares. O marcador RAPD foi mais discriminante em relação ao microssatélite pelo fato de agrupar os acessos em mais grupos. O banco de germoplasma da UFERSA possui alta variabilidade genética entre os acessos.

The high world energy demand requires a search for renewable sources which are less harmful to environment. Castor bean (Ricinus communis L.) cultivation is an excellent alternative to compound the national energy matrix due to its traditional small and intermediate producers, standing out as a culture with high economical and social appeal, especially in Northeast region. There is a lack of information, especially at molecular level, related to the amount of high genetic variability available in this oil crop. The objective of this work was to characterize the genetic diversity within ten genotypes of Ricinus communis L. by using RAPD molecular markers. A total of seven lines and three cultivars were evaluated, which included five lines from Germplasm Bank of Embrapa Algodão, two lines from Costa Rica and three Northeastern cultivars. Genomic DNA extraction was performed according both to Graham et al. (1994) and an alternative protocol proposed in this study. Seven arbitrary oligonucleotides employed in the RAPD-PCR amplification permitted to evaluate the efficiency of the genomic DNA extraction protocols, DNA banding profiles and calculation of similarity indexes. Similarity and cluster analysis were conducted using both Jaccard and Dice coefficient and the unweighted pair group method. The similarity data matrix was converted into a dendrogram. The amplified fragments yielded a total of 58 polymorphic bands ranging from 2394 to 3386 bp. On the basis of the performed analysis, similarity indexes and data clustering revealed the occurrence of high genetic variability among the genotypes. RAPD molecular markers were efficient on the detection of DNA polymorphism within genotypes evaluated, suggesting that this marker could be suitable for further characterization studies related to the main agroindustrial features of castor bean germplasm. / Fundação de Amparo a Pesquisa do Estado de Alagoas / A elevada demanda energética mundial faz necessária a busca por fontes renováveis e menos agressivas ao meio ambiente. O cultivo da mamona (Ricinus communis L.) constitui uma excelente alternativa na composição da matriz energética nacional por ser tradicionalmente praticado por pequenos e médios produtores, destacando-se como uma cultura de elevado apelo econômico e social, em especial na região Nordeste. Existe uma escassez de informações, especialmente no âmbito molecular, acerca da grande variabilidade genética apresentada por essa oleaginosa. O objetivo do presente trabalho foi caracterizar a diversidade genética entre dez genótipos de Ricinus communis L. por meio de marcadores moleculares RAPD. Foram avaliadas cinco linhagens do BAG da Embrapa Algodão, duas linhagens provenientes da Costa Rica e três variedades comerciais da região Nordeste. A extração do DNA genômico total dos genótipos foi realizada segundo metodologia descrita por Graham et al. (1994) e através de um protocolo alternativo proposto nesse estudo. O emprego de sete iniciadores arbitrários na amplificação RAPD-PCR permitiu avaliar a eficiência dos protocolos na extração do DNA genômico total, o padrão de bandas de DNA gerado e o cálculo de índices de similaridade. A matriz de dados de similaridade entre os genótipos foi convertida num dendrograma. Na análise dos fragmentos amplificados foram reveladas 58 bandas polimórficas com fragmentos de 2394 a 3386 pb. Ainda na avaliação dos resultados, a distância genética e o agrupamento gerado pelo dendrograma evidenciaram a grande variabilidade genética existente entre os indivíduos. Os marcadores moleculares RAPD mostraram-se eficientes na detecção de polimorfismos de DNA dos demais genótipos avaliados, tornando possível seu emprego em posteriores estudos de caracterização correlacionados com os principais caracteres agroindustriais de germoplasma de mamona.

Made available in DSpace on 2016-06-02T19:30:14Z (GMT). No. of bitstreams: 1 TeseRLC.pdf: 1717916 bytes, checksum: a24259cd3e84bb299a7f0093b50f8ad8 (MD5) Previous issue date: 2003-04-29 / Universidade Estadual de Maringá / Ilex paraguariensis, known as mate tea, is processed in many ways and commercialized as tea, soluble powder, essences, and processed leaves for preparation of mate tea without sugar ( chimarrão and tererê ). Aiming the conservation of this species, this work studied the intra- and interpopulational genetic variabilities of I. paraguariensis and its volatiles and semi-volatiles composition in 20 populations inside the area of incidence of this species in Brazil. Paternity tests were also performed to identify a favorable crossing for obtaining lineages with the characteristics of interest, reducing then the impact of extractive harvest on the native plants. Genetic variability of I. paraguariensis analyzed by RAPD markers (Random Amplified Polymorphic DNA) is lower to the reported variability in other dioic arboreal species. Intrapopulational variability is higher than interpopulational in this species. In intra and interpopulational studies alleles with the same allele sequence were identified, as well as alleles with very different sequences, what evidences the incidence of genetic flux among populations that are close geographically, but with alleles specific of each population. The environmental differences taken into account were not enough to explain genetic and chemical differences found among the populations. These results show the possibility of influences of microenvironmental factors and non-analyzed genes in the analyzed chemical composition of mate tea leaves. The population of Ponta Porã, geographically separated from the others, proved to be genetically distinct. Species-specific fragments were obtained for I. paraguariensis, I. dumosa and I. theezans. However, more studies for confirmation and utilization of such fragments must be carried out. The differences in the volatiles and semivolatiles composition among the populations turned their separation possible, and may be useful for identification and inclusion of genotypes, superior in specific compounds, in germoplasm collections. For conservation of this species in germoplasm collections ex situ, material collecting must be based on genetic analysis, as well as on geographical distribution, due to the higher intrapopulational variability and occurrence of rare alleles in the species. The RAPD technique may be used for paternity identification, mainly in arboreal species, making possible to obtain genetic improvement by performing controlled crossings. Based on these results, it was possible the discussion concerning the conservation aspects of this species. / Ilex paraguariensis, mais conhecida como erva-mate, é beneficiada por diversas formas e comercializada como chá, pó solúvel, essências e erva para chimarrão e tererê. Visando sua conservação, foram estudadas sua variabilidade genética intra e interpopulacional e sua composição química em 20 populações na área de ocorrência da espécie no Brasil. Também foram realizados testes de paternidade para identificação de um cruzamento favorável à obtenção de progênies com características de interesse, objetivando reduzir o impacto sobre ervais nativos causados pelo extrativismo. A variabilidade genética analisada por marcadores moleculares RAPD (polimorfismo de DNA amplificado ao acaso) em I. paraguariensis é inferior à variabilidade reportada em outras espécies arbóreas dióicas. A variabilidade intrapopulacional é maior que a variabilidade interpopulacional nesta espécie. Em estudos intra e interpopulacionais identificaram-se alelos com mesma freqüência alélica e alelos com freqüências bastante distintas, evidenciando a ocorrência de fluxo gênico entre populações próximas geograficamente, porém com alelos característicos de cada população. As diferenças ambientais consideradas não foram suficientes para explicar as diferenças genéticas e químicas encontradas entre as populações, demonstrando a possibilidade de haver interferências micro-ambientais e de genes ainda não analisados, influenciando na composição química da erva-mate. A população de Ponta Porã, separada geograficamente das demais, mostrou-se distinta geneticamente. Fragmentos espécie específicos foram obtidos para I. paraguariensis, I. dumosa e I. theezans, porém requerendo mais estudos neste sentido para confirmação e utilização destes. As diferenças nas análises dos compostos voláteis e semi-voláteis entre as populações, permitiram a separação destas e podem ser úteis para a identificação e inclusão de genótipos superiores em determinados compostos em bancos de germoplasma. Para a conservação desta espécie em bancos de germoplasma ex situ a coleta de materiais deve ser baseada em análises genéticas, além da distribuição geográfica, devido à maior variabilidade intrapopulacional e ocorrência de alelos raros na espécie. A técnica de RAPD pode ser utilizada para a identificação de paternidade, principalmente em espécies arbóreas, podendo-se obter ganhos genéticos com a condução de cruzamentos controlados. Os resultados obtidos embasaram discussões sobre aspectos de conservação desta espécie.

Made available in DSpace on 2016-12-08T16:44:39Z (GMT). No. of bitstreams: 1 PGPV11MA080.pdf: 299920 bytes, checksum: 31fddfd15b77ee0da63e2e67e528bf3f (MD5) Previous issue date: 2011-05-20 / Considering that genetic variability is essential for any breeding program, one of the first steps to be taken by the breeder is to determine the genetic variability available. Is this study sought to determine teh divergence between the genotypes studied using predictive techniques, ie, those based on the morphological, physiological or molecular, quantified in a measure of similarity or dissimilarity, wich can express the degree of genetic diversity among the possible parentes. We studied fifteen genotypes used in the state of Santa Catarina: Super superprecoce, Bela Vista, Baia Indaial, Crioula Roxa and Crioula Branca (Populations); Empasc 352 Bola Precoce, Empasc 355 Juporanga, Epagri 362 Crioula Alto Vale and Epagri 363 Superprecoce (comercial populations Epagri) bela Catarina, Bella Vista and Bella Dura (Hybrid s comercial Sakata) Boreal and Gauchinha (Hortec Populations company s business); Catarina (population Agritu comercial company). The first part of the work was divided into two experiments were conducted in experimental stations of Epagri Ituporanga and Lages. The design was a randomized block designs with three replications at each location. Weevaluated the following characteristics: i) length of pseudostem in cm, ii) number of leaves per pseudostem iii) stem diameter in mm; iv) bulb diameter in cm, v) height of the bulb in cm; vi) weight bulb in grams; vii) ratio of height: diameter of the bulb (value obtained by the ratio between height and diameter of the bulb), viii) total production of bulbs in kg há-1, ix) bulkb shape, x) flowering percentage and xi) percentage of rotten bulbs. We performed multivariate analysis of variance and produced a matrix of dissimilarity based on Mahalanobis distance. The variables that contributed most to the divergence were the production of bulbs and total length of the pseudostem to the environment Ituporanga and percentage of flowering and bulb weight of the environment Lages. The second experimente was conducted at the Institute Molecular genetics and Breeding of UDESC IMEGEM. Genotype were planted in the greenhouse, thus, the extraction of DNA from plants are Young. We used eleven primers of 10 bases, which produce thirty and five bands, wirh twenty-eight polymorphic. We used the Jaccard index as a measure of similarity and UPGMA clustering method for the preparation of a dendrogram of genetic similarity. The results showed that there is divergence between the genotypes studied, Crioula Roxa and Bola Precoce the most divergente, with 0.27 similarity and were less divergente Bella Vista and Bella Dura wirh 0.89 similarity / Considerando que a variabilidade genética é essencial para qualquer programa de melhoramento, um dos primeiros passos a ser dado pelo melhorista, é determinar a variabilidade disponível. Neste trabalho procurou-se determinar a divergência existente entre os genótipos estudados através de técnicas preditivas, ou seja, que têm por base as diferenças morfológicas, fisiológicas ou moleculares, quantificadas em uma medida de dissimilaridade ou similaridade, que possa expressar o grau de diversidade genética entre os possíveis genitores. Foram estudados quinze genótipos utilizados no estado de Santa Catarina: Super Superprecoce, Bela Vista, Baia Indaial, Crioula Roxa e Crioula Branca (Populações); Empasc 352 - Bola Precoce, Empasc 355 - Juporanga Epagri 362 Crioula Alto Vale e Epagri 363 - Superprecoce (Populações comerciais da Epagri); Bella Catarina, Bella Vista e Bella Dura (Híbridos comerciais da empresa Sakata); Boreal e Gauchinha (Populações comerciais da empresa Hortec); Catarina (população comercial da empresa Agritu). A primeira parte do trabalho foi dividida em dois experimentos, sendo realizados nas estações experimentais da Epagri de Ituporanga e de Lages. O delineamento utilizado foi de blocos casualizados com três repetições em cada local. Foram avaliadas as seguintes características: i) comprimento do pseudocaule em cm; ii) número de folhas por pseudocaule; iii) diâmetro do pseudocaule em mm; iv) diâmetro do bulbo em cm; v) altura do bulbo em cm; vi) peso do bulbo em gramas; vii) relação altura: diâmetro do bulbo (valor obtido pela razão entre a altura do bulbo e o diâmetro); viii) produção total de bulbos em kg ha-1; ix) forma do bulbo; x) porcentagem de florescimento e xi) porcentagem de bulbos podres. Foi realizada a análise da variância multivariada e elaborada uma matriz de dissimilaridade, com base na distância de Mahalanobis. As variáveis que mais contribuíram para a divergência foram a produção total de bulbos e comprimento do pseudocaule para o ambiente Ituporanga e percentagem de florescimento e peso do bulbo para o ambiente Lages. O segundo experimento foi realizado no Instituto de Melhoramento e Genética Molecular da UDESC IMEGEM. Os genótipos foram semeados em casa de vegetação, sendo realizada a extração de DNA das plantas ainda jovens. Foram utilizados onze oligonucleotídeos iniciadores de 10 bases, que produziram trinta e cinco bandas, sendo vinte e oito polimórficas. Foi utilizado o coeficiente de Jaccard como medida de similaridade e o método de agrupamento UPGMA para a elaboração de um dendrograma de similaridade genética. Os resultados demonstraram que existe divergência entre os genótipos estudados, sendo Crioula Roxa e Bola Precoce os mais divergentes, com 0,27 de similaridade e os menos divergentes foram Bella Vista e Bella Dura, com 0,89 de similaridade