Orientadores: Anete Pereira de Souza, Antonio Augusto Franco Garcia / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-08T00:10:19Z (GMT). No. of bitstreams: 1 Oliveira_KarineMiranda_D.pdf: 6932327 bytes, checksum: 92b9fb104f8543695a1c804fd65bf912 (MD5) Previous issue date: 2006 / Resumo: A cana-de-açúcar (Saccharum spp.) está entre as espécies de maior importância econômica no mundo, constituindo uma das principais fontes de produção de açúcar e álcool. Apesar do Brasil ocupar posição de destaque, como o maior produtor mundial, os níveis de produtividade são considerados baixos. Os programas de melhoramento para obtenção de novas variedades de cana-de-açúcar, mais produtivas e resistentes a pragas e doenças, podem ser acelerados com o desenvolvimento de marcadores genéticos, PCR específicos, fortemente ligados a genes que controlam as características de interesse. A utilização de marcadores em estudos de mapeamento genético e de QTL¿s (Quantitative Trait Loci) tem proporcionado um importante progresso no conhecimento da estrutura genômica e na genética da cana-de-açúcar. Sabe-se que os ESTs (Expressed Sequence Tags) apresentam grande potencial para serem utilizados com tais finalidades. O projeto de seqüenciamento de ESTs (SUCEST), do programa Genoma da FAPESP, identificou cerca de 43 mil clusters que representam os genes de cana-de-açúcar, potenciais para serem utilizados no desenvolvimento de marcadores genéticos. Neste contexto, o presente trabalho teve como objetivo desenvolver e mapear marcadores EST-SSRs em uma progênie derivada do cruzamento entre duas variedades pré-comerciais de cana-de-açúcar, complementando os programas de mapeamento genético desta população. Uma busca no banco de dados do SUCEST resultou na identificação de marcadores microssatélites ou SSRs (Single sequence repeats) em 2005 clusters. Trezentos e setenta e dois locos EST-SSRs foram desenvolvidos e analisados e, destes, 149 foram selecionados para estudo de mapeamento genético. Um total de 2303 marcadores polimórficos (SSRs, EST-SSRs, RFLPs, EST-RFLPs e AFLPs) foi identificado nos 100 indivíduos da progênie F1, dos quais 1669 (72%) eram marcadores em dose simples (1:1 e 3:1), segregantes na população. As análises de mapeamento foram baseadas na metodologia de identificação de marcadores em dosagem única no genoma, com o auxílio dos programas JoinMap (versão 3.0) e OneMap. Para a formação dos grupos de co-segregação (GCs) utilizou-se LOD 5 e fração de recombinação de 0.35. A função de mapeamento de Kosambi foi adotada para conversão das freqüências de recombinação em distâncias de mapa em centiMorgans (cM). Dos 1669 marcadores segregantes, 664 (40%) foram distribuídos em 192GCs, gerando um mapa com 6261.1 cM de comprimento. Os 192 GCs foram agrupados em 14 prováveis grupos de homologia. Cento e treze dos 149 EST-SSRs avaliados foram mapeados e apresentaram homologia a genes conhecidos de outras espécies. A adição dos marcadores provenientes de seqüências expressas, aumentou a cobertura e densidade do mapa prévio desta população, possibilitando também a construção do primeiro mapa funcional de cana-de-açúcar. Os EST-SSRs desenvolvidos têm potencial de utilização na detecção de QTL¿s associados a características de importância econômica para a cultura da cana-de-açúcar / Abstract: Sugarcane (Saccharum spp.) is one of the most important cash crops, highly contributing towards the production of raw sugar and bioethanol produced worldwide. Even though Brazil is the major producer, sugar yield is considered low. Breeding programs for the attainment of new improved sugarcane varieties, which are more productive and more resistant to plagues and illnesses, could be sped up with the development of genetic markers that are PCR-specific and strongly linked to genes that control the desired agronomic traits. The use of genetic markers in genetic mapping and QTL (Quantitative Trait Loci) studies has allowed important progress in the knowledge of the genomic structure and genetics of sugarcane. It is a known fact that the ESTs (Expressed Sequence Tags) have great potential to be used with such purposes. The Sugarcane Expressed Sequence Tag Project (SUCEST) has identified about 43000 clusters that represent the sugarcane genes with a potential to be used in the development of genetic markers. In this context, the objective of the present work was to develop and map EST-SSR markers in a progeny obtained by the cross between two pre-commercial sugarcane varieties, complementing the genetic mapping programs of this population. A search in the SUCEST database resulted on the identification of the microssatellites or SSR (Single sequence repeats) markers in 2005 clusters. Thus, three hundred and seventy two EST-SSR loci had been developed and analyzed and, out of these, 149 were selected for mapping analyses. A total of 2303 polymorphic markers (SSRs, EST-SSRs, RFLPs, EST-RFLPs and AFLPs) were identified among the 100 F1 individuals, of which 1669 (72.5%) were single dose (SD ¿ segregated 1:1 and 3:1) markers. Map analyses were carried out using JoinMap (version 3.0) and OneMap algorithm, based on a single-dose marker approach. The linkage relationships of simplex markers were determined at a LOD score threshold of 5 and a recombination fraction threshold of 0.35 and map distances were derived from the recombination fraction using the Kosambi function. Out of these 1669 SD markers, 664 (40%) were scattered onto 192 co-segregation groups (CGs) with a total estimated map length of 6261.1 cM. Using both genomic and EST-derived SSR and RFLP, 120 of the 192 CGs were formed into fourteen putative homology groups (HGs). One hundred and thirteen of the 149 EST-SSRs evaluated were mapped and presented homology to previously studied genes of other species. The addition of the EST-derived markers increased the coverage and density of the previous map of this population, which also enabled the construction of the first functional linkage sugarcane map. The EST-SSRs developed have the potential to be used in the detection of QTLs associated to important economic traits for sugarcane culture / Doutorado / Genetica Vegetal e Melhoramento / Doutor em Genetica e Biologia Molecular

O custo ferrugem que engloba gastos com fungicidas, os custos operacionais das aplicações, perdas de produção, e redução na arrecadação de impostos representa um elevado valor para sojicultura mundial. Assim, a ferrugem asiática da soja apresenta-se como um dos grandes desafios para a pesquisa agrícola nos próximos anos, pois o controle químico da doença é alternativa obrigatória enquanto não se dispõe de cultivares resistentes. Porém a prática representa aumento considerável no custo de produção. No Brasil estima-se que este custo, para duas aplicações gire, em torno de US$ 40,00.ha-1, sem contar os gastos operacionais com máquinas e combustível, ou seja, gasto apenas o fungicida. Considerando o exposto, este trabalho tem como objetivo avaliar duas fontes de resistência à ferrugem asiática da soja (PI 200487 e PI 459025A), e verificar se os genes responsáveis pela reação ao patógeno já foram descritos na literatura, ou seja, identificar seus respectivos grupos de ligação. A estratégia de segregantes agrupados (Bulked segregant analysis) foi utilizada para mapear os genes de interesse. Os resultados obtidos permitiram chegar as seguintes conclusões: a) a técnica BSA foi eficiente para detectar locos relacionados com a resistência à ferrugem asiática da soja; b) a PI 459025 (Bing Nam) apresenta o gene de resistência Rpp4 localizado no grupo de ligação G e c) A PI 200487 (Kinoshita) apresenta um gene diferente daqueles descritos na literatura, sendo este localizado no grupo de ligação N. Este gene pode ser denominado de Rpp5. / The costs due to Asian soybean rust, which consists of fungicides, operational application costs and reduction in paid taxes, represent severe losses for worldwide soybean production. Thus, Asian soybean rust represents one of the most challenging problems for agricultural research in the future, since chemical control of the disease is the obligated alternative while resistant cultivars are unavailable. However, fungicide application significantly increases production costs. In Brazil, it is estimated that the costs of two applications are of approximately US$ 40,00.ha-1, aside the operational costs incurred due to machinery and fuel, that is, the figure represents solely fungicide costs. Taking into account the previous considerations, the present work aims to evaluate two sources of resistance to Asian soybean rust (PI 200487 and PI 459025A), and to verify if the genes involved in pathogen-responses have already been described in previous work, by identifying their respective linkage groups. Bulked segregant analysis was used to map the genes of interest. The results allowed the following conclusions: a) BSA was effective to detect loci associated to Asian soybean rust resistance; b) PI 459025A (Bing Nam) carries the resistance gene Rpp4 mapped to the linkage group G and c) PI 200487 (Kinoshita) has a novel resistance gene mapped to the linkage group N. The identified gene is denominated Rpp5.

Made available in DSpace on 2016-08-12T19:18:22Z (GMT). No. of bitstreams: 1 FernandoASA_TESE.pdf: 1416076 bytes, checksum: b1c98d86beac7a63692f0bd1fd27af60 (MD5) Previous issue date: 2010-04-29 / The Brazilian melon agribusiness is in expansion, reaching almost 500 thousand tons per year and making melon the main domestic fruit in both exported volume and value, with this activity being concentrated in the Brazilian Semiarid region. However, Brazil ranks only twelfth among the producing countries in area planted and production and the twentieth-third in productivity, demonstrating that there is still much to do on melon breeding and genetic resources of this crop. Thus, the objectives of this work were to estimate the genetic diversity of melon accessions collected from the traditional agricultural areas of the Brazilian Northeastern region by using descriptors for the fruit and microsatellite markers, and to study the nature of genotype vs. environment interaction and to estimate genetic parameters and gains by selection of melon families, in the Jaguaribe/Assú irrigation projects (Ceará and Rio Grande do Norte States, Brazil). The study of genetic diversity showed great variability among accessions, within and among the botanical groups, enabling advances in genetic improvement of fruit quality traits. Cluster analysis by fruit descriptors formed eight groups, without taxonomic criteria. The characters fruit longitudinal diameter, number of fruits per plant, cavity thickness and total soluble solids were descriptors that contributed most to the dissimilarity of the genotypes. The SSR markers amplified 41 alleles with average of 2.41 alleles and three genotypes per locus. A phylogenetic molecular analysis separated the accessions into 13 groups, also without taxonomic coherence in groups formed. The degree of association between genetic distance morpho-agronomic and molecular matrices was null. Then, there was no significant association between the descriptors and SSR markers, which was predictable due the number of distinct groups, the quantitative nature of the descriptors and the distribution of SSR markers in the genome. In the study of genotype vs. environment interaction, there was heterogeneity among the families for all traits. Heritability estimates in the combined analysis were consistently lower than those estimated for each environment. The simple part of G vs. E interaction was always above 99%, except for pulp firmness, with a predominance of the large complex part between the pairs of environments A1 and A3 and A2 and A3. The direct gains by selection were higher than the indirect gains for all traits in all environments evaluated. Gains were closer to the direct genetic gains when the selection was based on the average of the three environments. Estimates of genetic parameters associated with the simple nature of the family vs. environment interaction allowed genetic progress by simple breeding methods. As the genotype-environment interaction overestimates the coefficient of genetic variation and genetic variance among families in an environment, genotype assessments in more than one location are necessary since that selection is made considering the average of the families on the environments. Therefore, the potential of genetic improvement was demonstrated for the germplasm used in the study of genetic diversity as well as for the germplasm evaluated in the analysis of genotype-environment interaction. / O agronegócio do melão brasileiro tem se expandido bastante, alcançando quase 500 mil toneladas de frutos por ano e tornando o melão a principal fruta nacional, tanto em volume de exportação quanto em valor exportado, estando esta atividade concentrada no semiárido brasileiro. Contudo, o Brasil é apenas o décimo segundo país em produção e área plantada e vigésimo terceiro em produtividade, evidenciando que ainda existe muito por ser feito em termos do melhoramento e recursos genéticos desta cultura. Assim sendo, os objetivos deste trabalho foram estimar a divergência genética de acessos de meloeiro coletados em propriedades de agricultura tradicional do Nordeste brasileiro, tanto por descritores do fruto quanto por marcadores microssatélites e estudar a natureza da interação genótipo x ambiente (G x A) bem como estimar parâmetros genéticos e ganhos por seleção de famílias de melão, no polo Jaguaribe-CE/Assu-RN. O estudo de divergência genética mostrou grande variabilidade entre os acessos, entre e dentro dos grupos botânicos, possibilitando avanços no melhoramento genético dos caracteres de qualidade dos frutos. A análise de agrupamento por descritores do fruto formou oito grupos, sem critérios taxonômicos. Os caracteres diâmetro lateral, número de frutos por planta, espessura da cavidade e sólidos solúveis foram os descritores que mais contribuíram para a dissimilaridade dos genótipos. Os marcadores SSR amplificaram 41 alelos com média de 2,41 alelos e três genótipos por loco. A análise filogenética molecular separou os acessos em 13 grupos, também sem coerência taxonômica nos grupos formados. O grau de associação entre as matrizes de distâncias genéticas morfoagronômica e molecular foi nulo, não havendo associação entre os descritores e os marcadores SSR, o que era previsível devido ao número distinto de grupos, a natureza quantitativa dos descritores e a forma de distribuição dos marcadores SSR no genoma. No estudo de interação genótipo x ambiente, houve heterogeneidade entre as famílias para todas as características avaliadas. As herdabilidades estimadas nas análises conjuntas foram sempre inferiores àquelas estimadas em cada ambiente. A parte simples da interação G x A foi sempre superior a 99%, exceto para firmeza da polpa, com amplo predomínio da parte complexa entre pares de ambientes A1 e A3 e A2 e A3. Os ganhos diretos com seleção foram maiores do que os ganhos indiretos para todas as características, em todos os ambientes avaliados. Os ganhos foram mais próximos dos ganhos genéticos diretos quando se praticou a seleção pela média dos três ambientes. As estimativas de parâmetros genéticos associadas à natureza simples da interação família x ambiente permitem progresso genético por meio de métodos de melhoramento menos sofisticados. Como a interação genótipo x ambiente superestima o coeficiente de variação genético e a variância genética entre famílias em um ambiente, são necessárias avaliações dos genótipos em mais de um local, desde que a seleção seja praticada considerando a média das famílias nos ambientes. Portanto, ficou comprovado o potencial de melhoramento genético tanto no germoplasma utilizado no estudo de divergência genética quanto do germoplasma avaliado nas análises da interação genótipo x ambiente.

Made available in DSpace on 2016-08-12T19:18:31Z (GMT). No. of bitstreams: 1 AnaCAD_TESE.pdf: 1798262 bytes, checksum: 234cf9c328c4c92a52de25fe960cac0a (MD5) Previous issue date: 2015-02-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The quality of the melon is one of the main objectives of breeding programs. An important aspect for the breeder is to know its inheritance as it assists in choosing the breeding strategy with time and resource savings. In addition, the use of biotechnological tools to assist in the knowledge of architecture of the characters and facilitate the selection process may be useful. The objectives of this study were: to identify SSR markers linked to QTLs of six characters related to the quality of melon and to study the genetic control of traits related to fruit quality of melon through classic study of generations involving mixed models. Based on this, we evaluated 100 F2: 3 families of crossing between the parents 'Hales Best Jumbo' (HBJ) and 'MR-1', momordica type, in the years 2013 and 2014. For the phenotypic evaluation of the families, two experiments were carried out in randomized block design with three replications. Each plot consisted of a line with five plants, spaced 2.0 m between rows and 0.3 m between plants. The traits evaluated were: fruit weight, shape index, flesh thickness, flesh firmness, soluble solids and percentage of fruit cracking. From the results, the identification of stable molecular markers linked to quality traits of melon fruit through multiple regression procedure was possible. Using multiple regression process were identified markers associated to the average fruit weight characters, shape index, flesh thickness, firmness, soluble solids and percentage of fruit cracking. The CMMS22-2, CMCTN4 and CMMS4-3 markers were the most stable in both evaluations. The CMGAN3 markers, CMAGN33, CMSTN4, CMMS4-3, CMMS22-2 were those who stood out as the most promising for assisted selection to improve the melon quality. For the inheritance study, the generations F1, F2 and backcross of crossings OL x A-16 and OL-PV were evaluated in two separate experiments carried out in randomized block design with three replications, in 2012. The traits evaluated were: average fruit weight, shape index, flesh thickness, flesh firmness and soluble solids. The analyses were performed using mixed models to estimate the parameters of the additive-dominant model and the variance components. The heritability in the narrow and broad sense and number of loci that control the character were estimated. Maximum likelihood hierarchical genetic models were used in tests of hypothesis in order to assess the presence of polygenes and major effect gene in the genetic control character. From the results, it is concluded that the inheritance of the studied characters is complex, with the presence of major gene and polygenes with additive and dominance effects / A qualidade dos frutos é um dos principais objetivos dos programas de melhoramento genético do meloeiro. Um aspecto importante para o melhorista é conhecer a herança do caráter, de vez que auxilia o pesquisador na escolha da estratégia de melhoramento com economia de tempo e recursos. Além disso, o uso de ferramentas biotecnológicas auxilia no conhecimento da arquitetura dos caracteres e facilita o processo seletivo. Os objetivos do presente trabalho foram: identificar marcadores SSR ligados aos locos gênicos (Quantitative Trait Loci - QTLs) controladores de seis caracteres relacionados à qualidade de frutos de meloeiro e estudar o controle genético de caracteres relacionados à qualidade de frutos de melão por meio de estudo clássico de gerações envolvendo modelos mistos. Foram avaliadas 100 famílias F2:3 do cruzamento entre os genitores Hales Best Jumbo (HBJ), tipo cantaloupensis e MR-1 , tipo momordica, nos anos de 2013 e 2014. Para a avaliação fenotípica das famílias, foram conduzidos dois experimentos, nos quais foi utilizado o delineamento em blocos casualizados com três repetições. Cada parcela foi constituída por uma linha com cinco plantas, com espaçamento de 2,0 m entre as linhas e 0,3 m entre plantas. Foram avaliadas as características peso médio do fruto, índice de formato, espessura da polpa, firmeza da polpa, sólidos solúveis e porcentagem de rachadura do fruto. Para genotipagem, foram utilizados os marcadores moleculares microssatélites ou SSR. Foi possível a identificação de marcadores moleculares estáveis ligados a caracteres de qualidade de frutos de melão pelo processo de regressão múltipla. Foram identificados marcadores associados aos caracteres peso médio do fruto, índice de formato, espessura da polpa, firmeza da polpa, sólidos solúveis e porcentagem de rachadura do fruto. Os marcadores CMMS22-2, CMCTN4 e CMMS4-3 foram os mais estáveis nas duas avaliações. Os marcadores CMGAN3, CMAGN33, CMSTN4, CMMS4-3, CMMS22-2 são os mais promissores para seleção assistida, visando ao melhoramento da qualidade de frutos do meloeiro. Para o estudo de herança, foram avaliadas os genitores: OL, tipo cantaloupensis; PV, tipo não definido; A-16, tipo acidulus; as gerações F1, F2 e os retrocruzamentos dos OL x A-16 e OL x PV em dois ensaios distintos conduzidos em blocos casualizados com três repetições no ano de 2012. Os caracteres avaliados foram: peso médio de fruto, índice de formato, espessura da polpa, firmeza da polpa e sólidos solúveis. As análises foram feitas utilizando modelos mistos para estimar os parâmetros do modelo aditivo-dominante e os componentes de variância. Foram estimados os parâmetros de herdabilidades nos sentidos amplo e restrito e número de locos que controlam o caráter. Utilizaram-se testes de hipóteses de máxima verossimilhança de modelos genéticos hierárquicos para avaliar a presença de poligenes e gene de efeito maior no controle genético do caráter. Pelos resultados, conclui-se que a herança dos caracteres estudados é complexa, com a presença de gene de efeito maior e poligenes com efeitos aditivos e de dominância

Made available in DSpace on 2014-08-20T13:25:38Z (GMT). No. of bitstreams: 1 dissertacao_liane_thurow.pdf: 690839 bytes, checksum: 6d23772ac7732bb0c45028fd46e4bcf0 (MD5) Previous issue date: 2013-10-18 / Based on the advanced knowledge in genomic, marker-assisted selection shows up as a promising strategy especially to accelerate the development of new cultivars in perennial species, such as peach. In this context, this dissertation addressed the current situation and the prospects of using marker-assisted selection in peach breeding programs as well as to characterize the genetic variability and population structure of an association panel representative of peach germplasm available in Brazil through SSR analysis. Based on a literature review, it was seen that a high number of QTL (Quantitative Trait Loci) is identified in genetic maps of peach, but, despite the importance of the characters studied, the use of markerassisted selection has been incipient by breeding programs. By the other side, with the recent completion of the sequencing of the peach genome, the perspective is that the low use of molecular tools by peach breeding program will change in the coming years. In this trend, the genetic variability present in the association panel representative of peach germplasm in Brazil is quite relevant. On the other hand, this germplasm is structured into two subpopulations according to the fruit flesh type, melting or non-melting, with some genotypes being a mixture of both types of fruit flesh. Greater variability is available in the melting flesh group. These results show that the association panel is suitable for association studies given its relevant genetic variability. However, it should be taken into account in the association studies the genetic structure intrinsic to this germplasm to avoid the identification of false markertrait associations. / Com base no avanço do conhecimento de genômica a seleção assistida por marcadores mostra-se como uma estratégia bastante promissora especialmente para acelerar o desenvolvimento de novas cultivares em espécies perenes, como o pessegueiro. Nesse contexto, esta dissertação aborda a situação atual e as perspectivas do uso de seleção assistida por marcadores moleculares no melhoramento genético de pessegueiro, assim como caracteriza a variabilidade genética e a estrutura de população do painel associativo de pessegueiro representativo do germoplasma disponível no Brasil por meio da análise de 10 loci SSR (Simple Sequence Repeats). Com base na literatura, um elevado número de QTLs (Quantitative Trait Loci) está identificado em mapas genéticos de pessegueiro, mas, apesar da importância dos caracteres envolvidos, até o momento o uso de seleção assistida por marcadores tem sido incipiente pelos programas de melhoramento. Por outro lado, com a conclusão recente do sequenciamento do genoma do pessegueiro, a perspectiva é de que essa realidade mude nos próximos anos. Corroborando com essa tendência, o primeiro passo para a seleção assistida por marcadores moleculares é conhecer a variabilidade genética presente no germoplasma de pessegueiro disponível no Brasil. Nesse sentido, os resultados desse trabalho mostram grande variabilidade genética no germoplasma avaliado. Porém, esse germoplasma está estruturado em duas subpopulações bem definidas e agrupadas de acordo com o tipo de polpa, fundente e não-fundente, com alguns acessos apresentando mistura de ambos, entre os quais a maior variabilidade está disponível dentro do subgrupo fundente. Com base nos resultados infere-se que o germoplasma avaliado é adequado para estudos de mapeamento associativo, dada a sua expressiva variabilidade genética, mas deve ser levada em consideração a estrutura genética desse germplasma a fim de evitar a identificação de associações falso-positivas entre marcadores e fenótipos de interesse.

O custo ferrugem que engloba gastos com fungicidas, os custos operacionais das aplicações, perdas de produção, e redução na arrecadação de impostos representa um elevado valor para sojicultura mundial. Assim, a ferrugem asiática da soja apresenta-se como um dos grandes desafios para a pesquisa agrícola nos próximos anos, pois o controle químico da doença é alternativa obrigatória enquanto não se dispõe de cultivares resistentes. Porém a prática representa aumento considerável no custo de produção. No Brasil estima-se que este custo, para duas aplicações gire, em torno de US$ 40,00.ha-1, sem contar os gastos operacionais com máquinas e combustível, ou seja, gasto apenas o fungicida. Considerando o exposto, este trabalho tem como objetivo avaliar duas fontes de resistência à ferrugem asiática da soja (PI 200487 e PI 459025A), e verificar se os genes responsáveis pela reação ao patógeno já foram descritos na literatura, ou seja, identificar seus respectivos grupos de ligação. A estratégia de segregantes agrupados (Bulked segregant analysis) foi utilizada para mapear os genes de interesse. Os resultados obtidos permitiram chegar as seguintes conclusões: a) a técnica BSA foi eficiente para detectar locos relacionados com a resistência à ferrugem asiática da soja; b) a PI 459025 (Bing Nam) apresenta o gene de resistência Rpp4 localizado no grupo de ligação G e c) A PI 200487 (Kinoshita) apresenta um gene diferente daqueles descritos na literatura, sendo este localizado no grupo de ligação N. Este gene pode ser denominado de Rpp5. / The costs due to Asian soybean rust, which consists of fungicides, operational application costs and reduction in paid taxes, represent severe losses for worldwide soybean production. Thus, Asian soybean rust represents one of the most challenging problems for agricultural research in the future, since chemical control of the disease is the obligated alternative while resistant cultivars are unavailable. However, fungicide application significantly increases production costs. In Brazil, it is estimated that the costs of two applications are of approximately US$ 40,00.ha-1, aside the operational costs incurred due to machinery and fuel, that is, the figure represents solely fungicide costs. Taking into account the previous considerations, the present work aims to evaluate two sources of resistance to Asian soybean rust (PI 200487 and PI 459025A), and to verify if the genes involved in pathogen-responses have already been described in previous work, by identifying their respective linkage groups. Bulked segregant analysis was used to map the genes of interest. The results allowed the following conclusions: a) BSA was effective to detect loci associated to Asian soybean rust resistance; b) PI 459025A (Bing Nam) carries the resistance gene Rpp4 mapped to the linkage group G and c) PI 200487 (Kinoshita) has a novel resistance gene mapped to the linkage group N. The identified gene is denominated Rpp5.

Knowledge of the genetic variation of crop collections is essential for their efficient use in plant breeding programs. The Ethiopian Highland Maize Germplasm Collection Mission was launched throughout the highlands of Ethiopia in 1998 and 287 traditional maize accessions were collected from farmers’ fields. To date, no information was available on the morphological and genetic diversity in this important collection. Various molecular marker techniques and quantitative genetics approaches were applied to accurately unravel the extent of phenotypic and genetic diversity, to study patterns of morphological and molecular variation and to determine association of molecular markers with quantitative trait variation, with the view of designing a sound breeding program and management strategy for maize in the highlands of Ethiopia. The morphological study confirmed that traditional Ethiopian highland maize accessions contain large amounts of variation for agro-morphological traits. The broad trait diversity observed among the accessions suggested ample opportunities for the genetic improvement of the crop through selection directly from the accessions and/ or the development of inbred lines for a future hybrid program. Selection practices followed by local farmers are mostly consistent within agroecology and gave rise to morphologically distinct maize accessions in different agroecologies. This underscores the importance of considering farmers’ knowledge of diversity in the collection and evaluation of local accessions. The results of amplified fragment length polymorphism (AFLP) and microsatellite or simple sequence repeat (SSR) marker analyses showed that bulking leaf samples from 15 individual plants per out-bred accession is an effective means of producing representative profiles of individual plants, thereby reducing the cost of DNA extraction and subsequent marker analysis of open-pollinated varieties. Cluster analyses based on AFLP and SSR data showed that most of the accessions collected from the Northern agroecology were genetically distinct from the Western and Southern accessions suggesting that differentiation for adaptive traits for drought conditions may have occurred in the Northern accessions. However, there was very little genetic differentiation between the Western and Southern accessions suggesting gene flow between the two agroecologies and recent introduction of similar improved varieties in these agroecoogies . In both marker systems, high mean genetic diversity was observed among the traditional Ethiopian highland maize accessions. This is possibly due to (i) the continuous introduction of maize from abroad by different organizations; (ii) genetic variation generated through farmers management practices; and (iii) the presence of different environmental conditions in the highlands of Ethiopia to which local landraces may have been adapted. The correlation between the morphological dissimilarity matrix and the matrices of genetic dissimilarity based on SSR and AFLP markers were 0.43 and 0.39, respectively (p = 0.001 in both cases). The correlation between SSR and AFLP dissimilarity matrices was 0.67 (p = 0.001). These significant correlations indicate that the three independent sets of data likely reflect the same pattern of genetic diversity, and validate the use of the data to calculate the different diversity statistics for Ethiopian highland maize accessions. From this study, three groups of maize accessions with distinctive genetic profiles and morphological traits were identified that will be useful for future collection, conservation and breeding programs of maize for the highlands of Ethiopia. A pilot association study using SSR markers and quantitative trait variation indicated that molecular markers could be useful to identify genetic factors controlling earliness, tallness, grain yield and associated traits, which could be exploited by various breeding schemes. The analytical tools outlined in this dissertation can be a useful tool in managing genetic variation of open-pollinated crops and will aid in the conservation of unique genetic diversity. Production stability and global food security are linked to the conservation and exploitation of worldwide genetic resources and this research attempts to add to that body of knowledge. Copyright 2005, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. Please cite as follows: Beyene, YA 2005, Genetic analysis of traditional Ethiopian Highland Maize (Zea Mays l.) using molecular markers and morphological traits : implication for breeding and conservation, PhD thesis, University of Pretoria, Pretoria, viewed yymmdd < http://upetd.up.ac.za/thesis/available/etd-02212006-112610 / > / Thesis (PhD (Genetics))--University of Pretoria, 2005. / Genetics / unrestricted

Sunflower (Helianthus annuus L.) high oleic acid content lines differ from conventional sunflower by an increase in oleic acid (C18:1) content of more than 60%. The current sunflower cultivars under production in South Africa are standard sunflower with high levels of linoleic acid (C18:2). The aim of this study was to improve the quality of oil produced by local sunflower germplasm with respect to oleic acid through employing a marker-assisted breeding technique to facilitate and speed up the recovery of the high oleic acid allele into the background of the recurrent parent genome. Eleven sunflower breeding genotypes with high and low oleic acid traits were obtained from the Agricultural Research Council-Grain Crops Institute (ARC-GCI) in Potchefstroom. The breeding genotypes were phenotypically characterised based on their oleic and linoleic acid levels using gas chromatography. Results demonstrated that the average mean of oleic and linoleic acid contents in high oleic acid genotypes were 72% and 17% respectively, while the average mean of oleic acid and linoleic acid contents in wild type lines were 33.5 % and 54 % respectively. These results indicated a perfect negative correlation between the amount of oleic and linoleic acids possessed in high and low oleic acid genotypes (R2 = -99.16%). Sequence characterised amplified region (SCAR) markers were tested to ascertain if any of the ten available dominant FAD2-1 markers was segregating with the high oleic acid allele. Four dominant SCAR markers (FAD2-1F4/R1; FAD2-1F4/R2; FAD2-1F13/R1; FAD2-1F14/R2) were strongly associated with the high oleic acid trait (P< 0.001). With regard to the inheritance of the high oleic acid trait, 143 plants of the F2 segregating population derived from a cross between the high oleic acid parent (AP901-95-3-4-1) and low oleic acid parent (H55-9-2-1-1) were genotyped with the four SCAR markers to determine the genetic state concerning the high oleic acid gene (Ol). Results from a Chi square analysis of the observed frequencies of each dominant FAD2-1 marker locus in 143 F2 individuals indicated that the deviation from the expected ratio of 3:1 (high to low oleic acid) was not statistically significant (P< 0.95) from the observed segregation ratio. These results were consistent with the previous finding that an incomplete dominant gene governs sunflower high oleic acid. A multiplex assay of 78 Simple sequence repeat (SSR) markers was optimised and evaluated on 143 plants of the F2 population to determine suitable SSR markers that can be used in a marker-assisted background selection. Only 14 markers were suitable for marker-assisted background selection based on their high polymorphic information content, allele frequency and maximum allele numbers. In conclusion, this study demonstrated the potential of using SSR and SCAR marker systems as a breeding tool to characterise and speed up the selection process in marker-assisted backcross breeding. / Thesis (MSc (Botany))--North-West University, Potchefstroom Campus, 2013.

Sunflower (Helianthus annuus L.) high oleic acid content lines differ from conventional sunflower by an increase in oleic acid (C18:1) content of more than 60%. The current sunflower cultivars under production in South Africa are standard sunflower with high levels of linoleic acid (C18:2). The aim of this study was to improve the quality of oil produced by local sunflower germplasm with respect to oleic acid through employing a marker-assisted breeding technique to facilitate and speed up the recovery of the high oleic acid allele into the background of the recurrent parent genome. Eleven sunflower breeding genotypes with high and low oleic acid traits were obtained from the Agricultural Research Council-Grain Crops Institute (ARC-GCI) in Potchefstroom. The breeding genotypes were phenotypically characterised based on their oleic and linoleic acid levels using gas chromatography. Results demonstrated that the average mean of oleic and linoleic acid contents in high oleic acid genotypes were 72% and 17% respectively, while the average mean of oleic acid and linoleic acid contents in wild type lines were 33.5 % and 54 % respectively. These results indicated a perfect negative correlation between the amount of oleic and linoleic acids possessed in high and low oleic acid genotypes (R2 = -99.16%). Sequence characterised amplified region (SCAR) markers were tested to ascertain if any of the ten available dominant FAD2-1 markers was segregating with the high oleic acid allele. Four dominant SCAR markers (FAD2-1F4/R1; FAD2-1F4/R2; FAD2-1F13/R1; FAD2-1F14/R2) were strongly associated with the high oleic acid trait (P< 0.001). With regard to the inheritance of the high oleic acid trait, 143 plants of the F2 segregating population derived from a cross between the high oleic acid parent (AP901-95-3-4-1) and low oleic acid parent (H55-9-2-1-1) were genotyped with the four SCAR markers to determine the genetic state concerning the high oleic acid gene (Ol). Results from a Chi square analysis of the observed frequencies of each dominant FAD2-1 marker locus in 143 F2 individuals indicated that the deviation from the expected ratio of 3:1 (high to low oleic acid) was not statistically significant (P< 0.95) from the observed segregation ratio. These results were consistent with the previous finding that an incomplete dominant gene governs sunflower high oleic acid. A multiplex assay of 78 Simple sequence repeat (SSR) markers was optimised and evaluated on 143 plants of the F2 population to determine suitable SSR markers that can be used in a marker-assisted background selection. Only 14 markers were suitable for marker-assisted background selection based on their high polymorphic information content, allele frequency and maximum allele numbers. In conclusion, this study demonstrated the potential of using SSR and SCAR marker systems as a breeding tool to characterise and speed up the selection process in marker-assisted backcross breeding. / Thesis (MSc (Botany))--North-West University, Potchefstroom Campus, 2013.

Made available in DSpace on 2016-12-23T14:37:43Z (GMT). No. of bitstreams: 1 Sara Coser Morra.pdf: 1157047 bytes, checksum: ca26cc99cfaca69da596f91375dc0dcb (MD5) Previous issue date: 2012-07-12 / A goiabeira (Psidium guajava L.) é uma das fruteiras de maior importância econômica da família Myrtaceae. O Brasil é um dos maiores produtores de goiaba do mundo, sendo esta uma cultura potencial em expansão e rentabilidade. A polinização cruzada da espécie e a existência de pomares heterogêneos de propagação seminal resultam em variabilidade, permitindo a seleção de genótipos para o melhoramento da cultura. O objetivo deste trabalho foi estudar a diversidade genética em genótipos de P. guajava selecionados em pomar de origem seminal e cultivares por características morfológicas e químicas de qualidade de fruto, também associar dados de conteúdo de DNA nuclear (2C), cariótipo, morfológicos e de marcadores moleculares microssatélites. Verificou-se a existência de divergência entre as Cortibel com relação às características de fruto, com genótipos apresentando performance superior e genótipos com desempenho semelhante à cultivares, potenciais para uso em hibridações ou como cultivares. As análises cariotípica e de conteúdo de DNA nuclear (2C) mostraram que os genótipos possuem um genoma estável, pequeno e diplóide e características cariotípicas relacionadas a grupos ancestrais de angiospermas. O dendrograma UPGMA baseado em dados morfológicos e SSR evidenciaram diversidade entre os genótipos, com melhor discriminação pelos dados de SSR. Como a maioria dos genótipos mostraram similaridade morfológica para as características de frutos, aliada a dissimilaridade molecular, estes se mostraram interessantes para o uso em hibridações em programas de melhoramento. O conjunto de dados gerados contribuiu para expandir o conhecimento sobre o genoma e a diversidade genética em P. guajava. Também é importante na estruturação de programas de melhoramento para a cultura, além de contribuir para estudos evolutivos / Guava (Psidium guajava L.) is one of the most economically important fruit crop from Myrtaceae family. Brazil is one of the largest producers of guava in the world, which is a potential crop in growth and profitability. Cross-pollination of the species and the existence of heterogeneous seminal propagation orchards result in variability, allowing the selection of genotypes for crop improvement. The aim of this study was to evaluate genetic diversity between P. guajava L genotypes selected from seminal origin orchard and cultivars, by morphological and fruit quality chemical characteristics, also associate data from nuclear 2C-value, karyotypic, morphological and simple sequence repeat (SSR) marker. There were divergences between Cortibel selections by fruit characteristics, with genotypes showing superior and similar performance when compared with cultivated genotypes, potential for use in hibridation and as cultivars. Karyotype and nuclear 2C-value analyses showed that all genotypes have a stable and very small diploid genome (2n = 2X = 22; 2C = 0.95 pg), and karyotypic characteristics related to ancestral angiosperm groups. UPGMA dendrogram based on morphological and SSR data evidenced diversity among the genotypes, with better discrimination by SSR data. Since most genotypes showed morphological similarity for fruit characteristics, combined with molecular dissimilarity, the use of these genotypes in hybridation breeding programs could be of interest. The obtained data set contributed to expand the knowledge about genome and genetic diversity of P. guajava. Also are important to structure crop improvement programs and contribute to evolutionary approaches