Submitted by Nathália Faria da Silva (nathaliafsilva.ufv@gmail.com) on 2017-07-07T16:54:28Z No. of bitstreams: 1 texto completo.pdf: 697730 bytes, checksum: 97e65860ffb26826b1b8a3b89981adc8 (MD5) / Made available in DSpace on 2017-07-07T16:54:28Z (GMT). No. of bitstreams: 1 texto completo.pdf: 697730 bytes, checksum: 97e65860ffb26826b1b8a3b89981adc8 (MD5) Previous issue date: 2001-03-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo deste trabalho foi avaliar o efeito da temperatura e do tempo de molhamento foliar nos componentes epidemiológicos de isolados das linhagens clonais US-1 e BR-1 de isolados de Phytophthora infestans existentes no Brasil e a validação do simulador de epidemias de requeima Blight nas condições ambientais do País. Os componentes epidemiológicos avaliados neste estudo foram o período de incubação, o período latente, o crescimento das lesões, a esporulação e a germinação dos esporângios do patógeno. Estudou-se o efeito da temperatura na germinação dos esporângios, no período de incubação, no período latente, no crescimento das lesões e na esporulação do patógeno. As linhagens clonais US-1 e BR-1 foram afetadas de maneira diferenciada pelas variações de temperatura. Os isolados de US-1 e BR-1 apresentaram maior percentual de germinação indireta em temperaturas inferiores a 15°C. O maior percentual de germinação direta foi a 22°C, em ambas as linhagens. Os menores valores de período de incubação e de período latente ocorreram a 22°C em ambas as linhagens. A essa temperatura, o período de incubação para US-1 foi de 66,7 h e, para BR-1, de 44,0 h, enquanto o período latente foi de 93,3 h para US -1 e de 68,0 h para BR-1. O crescimento das lesões também foi influenciado pela variação de temperatura entre 10 e 27°C, em ambas as linhagens clonais. Na avaliação da esporulação, em plantas de batata e tomate observou-se maior esporulação a 22°C. Na quantificação dos efeitos do binômio temperatura – tempo de molhamento foliar, a linhagem clonal BR-1 apresentou maior número de lesões no hospedeiro a 10°C com 24 horas de molhamento foliar, e a US-1, a 15°C, com o mesmo tempo de molhamento foliar. Na validação do simulador Blight, realizaram-se dois ensaios de campo, um no inverno e outro no período de primavera-verão, com a cultura da batata. Quantificou-se o progresso da doença nessas duas épocas, e com os dados climáticos coletados foram realizadas simulações. Houve boa correlação entre a epidemia observada no campo no período do inverno e a simulada. No segundo ensaio, o simulador subestimou a intensidade da doença. Para ser usado no Brasil, o simulador Blight necessitará de ajuste nas equações matemáticas que o compõem. / This study aimed at evaluating temperature and leaf-wetness period effect on the epidemiological components of clonal lines of Phytophthora infestans isolates US-1 and BR-1 present in Brazil and to validate the simulator of the late blight epidemic Blight under Brazilian environmental conditions. The isolates were affected differently by the temperature variation although maximum indirect and direct sporangial germination of both the isolates occurred at temperatures below 15°C and at 22°C, respectively. Both the isolates had the least incubation and latent period at 22°C, which was, respectively, 66.7 h and 93.3 h for US-1 and 44.0 h and 68.0 h for BR-1. In both the clonal lines, the lesion growth was also affected by the temperature variation between 10 to 27°C. The maximum sporulation on potato and tomato plants occurred at 22°C. Quantification of the combined effect of temperature and leaf-wetness period showed that the largest lesion, in 24 h, on the host was produced at 10°C by the BR-1 and at 15°C and by the US-1 lines with 24 h leaf-wetness period. The Blight simulator was validated on the winter and spring-summer season potato crop . Simulations were done by quantifying the disease progress and using the climatic data of the crop period. While there was a good correlation between the observed epidemic and the simulation in the winter planting, the simulator under- estimated the disease intensity in the spring-summer season. Therefore, the Blight simulator may require adjustments in the component mathematical equations for use in Brazil.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2016-04-20T08:03:41Z No. of bitstreams: 1 texto completo.pdf: 351964 bytes, checksum: f91834fae1f8b64d3fed68307f2774b1 (MD5) / Made available in DSpace on 2016-04-20T08:03:42Z (GMT). No. of bitstreams: 1 texto completo.pdf: 351964 bytes, checksum: f91834fae1f8b64d3fed68307f2774b1 (MD5) Previous issue date: 2015-08-10 / A podridão radicular de fitóftora (PRF) é uma das doenças mais agressivas para a cultura da soja, causando danos que podem levar a morte das plantas. O uso de cultivares resistentes é a principal estratégia para redução das perdas causadas por Phytophthora sojae. Por este motivo, estudos de identificação de genes e QTLs (Quantitative Trait Loci) associados à resistência parcial à PRF e associação de marcadores moleculares ligados à estes são de extrema importância. Neste trabalho foi realizada a genotipagem ampla de 68 cultivares de soja utilizando 5353 marcadores SNPs (Single Nucleotide Polymorphism), objetivando encontrar marcadores associados a resistência parcial de soja à Phytophthora sojae através de metodologias de genética de associação. Como resultado, neste trabalho foram localizados via metodologia de modelos lineares mistos (MLM) 23 marcadores candidatos associados à resistência parcial à PRF, dispostos em 9 cromossomos e, dentre estes marcadores, destacou-se o Gm16_29528259_T_C, por ser o com maior valor de associação (-log10(p)=2,9175) e os marcadores Gm05_8656389_T_C e Gm05_8695812_A_G, que apresentaram os maiores valores de r3 (0,18769), explicando, individualmente, 18,769% da variação fenotípica. Via metodologia de stepwise foram localizados 5 marcadores candidatos associados à resistência parcial à PRF (Gm10_37469103_C_A, Gm07_5417760_T_C, Gm11_37106045_C_T, Gm09_41620640_G_T e Gm09_45586982_T_C), dispostos em 4 cromossomos. Estes marcadores juntos obtiveram alto valor de r2 (0,6884), explicando 68,84% da variação fenotípica. Os marcadores SNPs associados à resistência parcial à podridão radicular de fitóftora estão distribuídos em dez grupos de ligação, sendo que, os grupos O, M, B1, A1 e A2, ainda não foram descritos como portadores de regiões controladoras da resistência. Os marcadores candidatos associados à resistência parcial à PRF localizados neste trabalho, após devidamente validados, podem ser utilizados para estabelecer um eficiente programa de seleção assistida por marcadores moleculares para resistência parcial de soja à PRF. / Phytopthora root and stem rot (PRSR) is one of the most aggressive diseases for the soybean crop, causing damage that can cause plant death. The uses of resistant cultivars is the main strategy to reduce losses cause by Phytophthora sojae. For this reason, studies for identifying genes or QTLs (Quantitative Trait Loci) associated with the partial resistance between this disease and the association of molecular markers linked to these genes or QTLs are paramount. In this research was carried out wide genotyping of 68 soybean cultivars using 5353 SNPs (Single Nucleotide Polymorphism) markers. The objective of this research was to identify markers associated with partial resistance to Phytophthora sojae, in soybeans through genetic screening methodology. Were located in the mixed linear models (MLM) methodology 23 candidate markers associated with partial resistance to PRSR, arranged on 9 chromosomes. The SNP Gm16_29528259_T_C stands out for being the higher value of association (-log10(p)=2,9175), the SNPs Gm05_8656389_T_C and Gm05_8695812_A_G showed the highest r2 values (0,18769), explained individually 18,769% of the phenotypic variation. Using stepwise methodology 5 markers associated (Gm10_37469103_C_A, with partial resistance to Gm07_5417760_T_C, PRST were located Gm11_37106045_C_T, Gm09_41620640_G_T and Gm09_45586982_T_C), arranged on four chromosomes. These markers together achieve high r2 value (0,6884), explaining 68,84% of the phenotypic variation. SNPs markers associated with partial resistance to PRSR are spread in ten linkage groups, wherein then groups O, F, B1, A1 and A2 have not been described as having the resistance controlling regions. The candidates markers associated with partial resistance to PRSR located in this research, after being properly validated, can be used to establish and effective marker-assisted selection (MAS) program for partial resistance to PRSR in soybean. / Sem lattes e agência de fomento

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico. Programa de Pós-graduação em Engenharia de Produção / Made available in DSpace on 2012-10-20T21:06:49Z (GMT). No. of bitstreams: 1 225932.pdf: 1800466 bytes, checksum: 7546469656bbd6296a6ac29ff925f29e (MD5) / Este trabalho apresenta um estudo desenvolvido com dados longitudinais, usando a metodologia de modelos não lineares em curvas de crescimento, com diferentes estruturas para a matriz de covariância, fixando uma função para a parte determinística. Após a seleção da melhor matriz de covariância, foram experimentadas diferentes funções, a fim de se escolher o modelo não linear mais adequado. Este procedimento foi aplicado em dados da porcentagem de severidade da doença Late blight em quatro variedades de batata (Solanum tuberosum), causada por Phytophthora infestans. O modelo não linear selecionado foi o que usa uma das parametrizações da função de Gompertz e matriz de covariância de Simetria Composta. Os testes de hipóteses realizados sobre os parâmetros do modelo confirmaram a existência de diferença significativa entre as variedades.

Made available in DSpace on 2016-06-02T18:57:50Z (GMT). No. of bitstreams: 1 5940.pdf: 754678 bytes, checksum: d36d72758fb4a273690a2c5829a7ca95 (MD5) Previous issue date: 2014-05-29 / Financiadora de Estudos e Projetos / Among the phytosanitary problems faced by citrus sector it is highlighted the root rot caused by Phytophthora spp. which affects all stages of development of the citrus trees. Preventive measures associated with chemicals are used to control the disease, but the problems arising from the intensive use of fungicides have increased the interest in developing alternative techniques aimed at agricultural sustainability, among them the use of biofertilizers . Because of these facts, this study aimed to: (i) assess the effect of microbiota and metabolites present in biofertilizers produced from aerobic digestion (BSA) and anaerobic (BSAN) of swine manure on the mycelial growth of P. nicotianae; (ii) determining the effect of the products on inoculate of P. nicotinae in soil and roots of citrus tree, (iii) observing the effect on germination of citrus rootstocks . The results obtained in this study showed that the existing microorganisms and metabolites in biofertilizers affected the development of the colony of P. nicotianae; biofertilizer produced by aerobic digestion of swine manure (BSA) had the best efficiency in the elimination of inoculum of P. nicotianae ; BSA at concentrations of 5 and 10% and BSAN at concentration of 10% favored the germination of mandarin Sunki in substrate infested by the pathogen. Moreover BSA at concentration of 5% had a promising effect on seed germination of Rangpur lime under the same conditions. / Dentre os problemas fitossanitários enfrentados pelo setor citrícola, destacam-se doenças ocasionadas por Phytophthora spp. que afetam todos os estádios de desenvolvimento da planta cítrica. Para o controle dessas doenças são utilizadas medidas preventivas associadas com produtos químicos, porém, os problemas advindos do uso intensivo de fungicidas têm aumentado o interesse em desenvolver técnicas alternativas, visando a sustentabilidade agrícola e, dentre essas o uso de biofertilizante. Diante do exposto, o presente trabalho teve por objetivos: (i) avaliar o efeito da microbiota e dos metabólitos presentes em biofertilizantes, produzidos pela digestão aeróbica (BSA) e anaeróbica (BSAN) de esterco suíno, sobre o desenvolvimento micelial de P. nicotianae; (ii) verificar o efeito dos compostos sobre propágulos de P. nicotinae no solo e nas raízes de citros e, finalmente, (iii) observar o efeito na germinação de sementes de porta-enxertos de citros em substrato inoculado. Os resultados obtidos nesta pesquisa mostraram que os microrganismos e os metabólitos existentes nos biofertilizantes afetaram o desenvolvimento da colônia de P. nicotianae; o biofertilizante produzido pela digestão aeróbica de esterco suíno (BSA) foi o que apresentou melhor eficiência quanto à supressão de inóculo de P. nicotianae; BSA nas concentrações de 05 e 10% e BSAN a 10% favoreceram a germinação das sementes de tangerina Sunki, em substrato infestado pelo fitopatógeno; enquanto que, para o limão cravo, apenas BSA a 5% teve um efeito promissor na germinação das sementes nas mesmas condições.

2 p. / Alfalfa varieties differ in fall dormancy, defined as growth during the fall. Nondormant alfalfa varieties are usually planted in mild winter areas for their ability to grow in the fall. However, fall growth of nondormant alfalfa may be undesirable in areas subject to repeated frosts or freezes. Nondormant, very nondormant, and extremely nondormant alfalfa varieties (fall dormancy class 8, 9, and 10) are adapted to elevations below 4000 feet in Arizona. Other dormancy classes not included in this publication are moderately nondormant varieties (fall dormancy class 7) which may be grown from 3000 to 5000 feet, and semi-dormant and dormant varieties (fall dormancy 6 and below) which are adapted to colder winter areas above 4000 feet.

M.Sc. / in the current study the effect of the addition of methyl Jasmonate (MeJA), chitosan, a cell wall elicitor prepared from Phytophthora nicotlanae to tobacco cells and the subsequent defense responses elicited in these cells were Investigated. The defense responses investigated can be divided into three categories according to the time scale whereby resistance responses in plant cells are induced: early events which included the analysis of lipid peroxidation, the induction of lipoxygenase (L0)0 enzyme activity as well as the changes in phosphoprotein profiles; intermediate to later responses which included investigations of peroxidase (POD) activity, lignin content, phytoalexin content and phenolic content and also late responses which included studies of pathogenesis-related proteins (PR) and 13-1,3-giucanase activity. An approach also followed in this study was the addition of MeJA to tobacco cells for 24 h followed by the addition of either the cell wall elicitor or chitosan as a secondary elicitors, to investigate possible preconditioning or sensitisation by MeJA. Results obtained in this study revealed the time and concentration dependent accumulation of phytoalexins (secondary metabolites) when MeJA was added to tobacco cells and an optimal concentration of MeJA to use in further studies was determined as 1 mM. MeJA was the most effective inducer of lipid peroxidation (22 fold induction), a response observable after 2 h of exposure to MeJA. Conditioning with MeJA, followed by both chitosan (19 fold induction) and elicitor (25 fold induction) led to an earlier accumulation as well as significant increases in the levels of malondialdehyde, the product of lipid peroxidation. LOX enzyme activity was significantly increased by the addition of MeJA (6 fold Induction), chitosan (4 fold induction) and elicitor (3.8 fold induction). Conditioning with .MeJA, followed by both chitosan (3.3 fold induction) and elicitor (3.9 fold Induction) also led to noteworthy increases in enzyme activity. Analysis of the phosphoprotein profiles do not reveal the accumulation of phosphorylated proteins when MeJA was added to cells and very little accumulation of such proteins when chitosan was added. Phosphorylated proteins could be observed in cells treated with elicitor and In the cases where conditioning with MeJA, followed by secondary elicitation with either chitosan or elicitor, was studied, the differential induction of phosphorylated cellular proteins could also be observed. No significant induction of POD activity could be observed under any of the conditions, except for a possible slight increase in POD activity starting at 16 - 24 h after the elicitor had been added and a more definite increase after 24 h which was sustained up to 48 h after the addition of MeJA. PAGE of peroxidase, followed by activity staining revealed the presence of a slow migrating anionic peroxidase as well as a fast migrating peroxidase. Conditioning with MeJA, followed by secondary elicitation with both chitosan and elicitor revealed enhanced POD activity as well increased induction of a fast migrating anionic peroxidase on PAGE gels. MeJA was a more effective inducer of elevated levels of lignin content than the elicitor or chitosan and the addition of MeJA to tobacco cells led to a 2.2 fold increase in the lignin content, a response observed after 24 h and sustained up to 48 h. Chitosan as secondary elicitor did not lead to any increase in lignin content, but the cell wall elicitor as secondary agent significantly increased the lignin content after 40 - 48 h. Analysis of phenolic content did not show any significant increases In the total soluble phenolics when the agents were used on their own and only the phenolic content of the MeJA-conditioned cells, followed by the addition of chitosan showed a slight increase. In this case, the HPLC analysis of the phenolics also revealed a shift In the profiles for phenolics. SDS-PAGE of PR proteins revealed the induction of constitutive as well as new proteins when MeJA and elicitor, but not chitosan were used as elicitation agents. However, In the MeJA-pretreated cells addition of both chitosan and elicitor led to increased accumulation of PR proteins with molecular masses ranging from 6 - 70 kDa. Results from the i3-1,3-glucanase activity assay indicate a strong induction (4-5 fold) when MeJA and elicitor (4 fold), but not when chitosan was added to cells. Conditioning effects were revealed when both chitosan (3 fold induction) and elicitor (2.5 fold induction) were used as secondary elicitors. The increases in intensities of bands with molecular masses ranging from 31- 35 kDa observed on SOS-PAGE gels where chitosan and elicitor were added as secondary agents corresponded in a time dependent manner with the increased levels obtained in thep-1,3-glucanase activity assay.

Les oomycètes représentent un groupe de microorganismes eucaryotes filamenteux distincts phylogénétiquement des champignons incluant de nombreuses espèces phytopathogènes. CBEL est une glycoprotéine pariétale de Phytophthora parasitica constituée d'une répétition de deux régions séparées par un linker. Chaque région protéique est constituée d'un domaine protéique de liaison à la cellulose (CBM1) et un motif PAN /Apple impliqué dans des interactions protéines-protéines ou protéines-polysaccharides. Cette étude doctorale porte sur la caractérisation de la protéine CBEL et plus particulièrement de ses CBM1s ainsi que sur l'évaluation et optimisation du potentiel de cette protéine à : (i) stimuler les défenses naturelles des plantes (ii) augmenter l'activité de glycosides hydrolases. Dans la première partie de ce travail doctoral différents tests visant à reproduire un traitement éliciteur externe sur plante entière ont pour cela été développés. Ces tests ont permis de mettre en évidence que formulée en présence de surfactants CBEL est capable d'induire diverses réponses de défense chez A. thaliana. Une production en masse de cette protéine a été réalisée dans la levure Pichia pastoris et la bactérie Escherichia coli dans l'optique d'une future application agronomique. Les protéines recombinantes CBELcol et CBELpic produite dans ces différents systèmes d'expression présentent des profils de glycosylation différents de celui de la protéine native CBELnat. Alors que ces protéines semblent se lier de manière identique à la cellulose les différents tests d'élicitation développés au cours de ce travail mettent en évidence des variations dans leur activité élicitrice suggérant que la nature des résidus glucidiques présents sur cette glycoprotéine peut avoir un impact sur sa capacité induire des réponses de défenses en application externe. Lors de la deuxième partie de ce travail de thèse la capacité de CBEL à interagir avec différents substrats cellulosiques a été caractérisée. Les résultats obtenus ont permis de montrer que CBEL se lie avec une haute affinité à la cellulose cristalline avicel et que la présence de CBM1 fonctionnels est nécessaire à cette interaction. De manière intéressante, le CBM1-1 et CBM1-2 ne semblent pas contribuer de manière égale à cette interaction. Par ailleurs la laison de CBEL à la cellulose induit des perturbations structurales sur le substrat et permet d'améliorer l'activité de la xylanase XynB de Talaromyces versatilis sur paille de blé. En outre une xylanase chimère possédant dans sa séquence le CBM1-1 de CBEL possède également une activité augmentée sur paille blé. L'ensemble de ces résultats met en évidence le potentiel de CBEL et de son CBM1-1 pour l'amélioration de l'activité de glycoside hydrolases utilisables par exemple en bioraffinerie. En dernier lieu un travail de caractérisation structurale de la protéine CBEL a également été entamé au cours de cette étude. L'enveloppe de la protéine CBEL en solution à notamment été déterminée par SAXS (Small Angle X-ray Scattering) et un modèle 3D de cette protéine a été obtenu. / Oomycetes are fungal like microorganisms evolutionary distinct from true fungi that include pathogens of plants. CBEL is a cell wall glycoprotein isolated from the oomycete Phytophthora parasitica that is composed of two distinct regions linked by a threonine/proline rich linker. Each region owns a cellulose binding module (CBM1) and a PAN-Apple domain involved in protein-protein or proteins-polysaccharides interactions. Since CBEL is able to induce defense responses in numerous plant species, its use for the development of products able to protect crops has been envisaged. For this purpose we analysed the effect of an external CBEL treatment on plants. We found that in the presence of surfactants CBEL is able to induce cytosolic calcium changes, defense gene expression, and cell death on A. thaliana. CBEL application for crop protection requires the development of economically reliable production processes. In the case of proteinaceous elicitors, an attractive strategy to obtain large amount of elicitors is to express them in heterologous hosts such as bacteria or yeasts. CBELcol and CBELpic were produced respectively in E. coli and in P. pastoris. CBELcol is unglycosylated whereas CBELpic displays a glycosylation profile distinct from the native protein (CBELnat). We found that all these proteins are able to bind crystalline cellulose. On the other side we found that the elicitor activity of CBELpic is distinct from CBELnat and CBELcol suggesting that the glycosylation on CBEL can have an impact on its ability to induce plant defense responses after external treatment on A. thaliana. In the second part of this work the two CBMs (1-1 and 1-2) that form part of CBEL have been submitted to detailed characterization, first to better quantify their interaction with cellulose and second to determine whether these CBMs can be useful for biotechnological applications, such as biomass hydrolysis. A variety of biophysical techniques were used to study the interaction of the CBMs with various substrates and the data obtained clearly indicate that CBEL's CBM1-1 exhibits much greater cellulose binding ability than CBM1-2. Engineering of the family 11 xylanase from Talaromyces versatilis (TvXynB), an enzyme that naturally bears a family 1 CBM, has produced two variants. The first one lacks a CBM, whereas the second contains the CBEL CBM1-1 in the place of the natural CBM1. The study of these enzymes has revealed that wild type TvXynB binds to cellulose, probably via its CBM1, and that the substitution of its CBM by oomycetal CBM1-1 does not affect its activity on this substrate. Moreover, the presence of CBEL during the hydrolysis of wheat straw actually potentiates the action of TvXynB, a result that is consistent with the hypothesis that CBM1-1 can alter cellulose surface fibres rather like some other members of CBM family 1.

During the initiation and execution of a rootstock breeding programme to overcome the financially crippling disease, Phytophthora root rot of avocado, various constraints have been identified for both the breeding as well as the screening aspect of the programme. A review of the literature revealed a complex host-pathogen interaction that should be taken into account in the recombination and screening of genetic material. With the detection of beneficial genotypes being the crux of a breeding programme, this dissertation was focused on the screening of rootstock material for tolerance to Phytophthora cinnamomi. Screening should be scientific but at the same time also be time and cost effective. Specific attention was given to (i) the correct medium for screening mass numbers of seedlings, (ii) fast and effective cloning of single selections, and (iii) evaluation of clonal material for tolerance to P. cinnamomi. Soil as a screening medium was compared with three inert hydroponic media as well as one aeroponic system. Only soil was found to be ineffective due to its properties. The other media tested, namely, sand, vermiculite, water and the aeroponic system were equal in performance. The medium to be used will depend on the preference of the breeder as each medium has its own pro's and con's. It was, however, found that the evaluation criterion to be applied depends on the medium that is used. With regard to cloning of single selections, a definite difference with regard to the cloning ability of the different selections was found. An inability to be etiolated was displayed by some of the selections and these could thus not be vegetatively propagated and were not further tested. One of the tolerance mechanisms in the standard cultivar Duke 7, is root regeneration. It was thus expected that this characteristic cloning would give an indication of the rootstock's ability to tolerate P. cinnamomi. This could not be confirmed, but most of the selections did, however, perform better than Duke 7. Comparison of feeder root percentage in non-inoculated and inoculated treatments was not sufficient for facilitating the final selection of candidate rootstocks from a large number of potential clonal selections. Four selections were made, based on the hypothesis that a larger root system will be a better forager and thus enhance the horticultural aspects of the rootstock-scion combination. Valuable information was obtained with regard to various mediums and criteria to be used during mass screening and final screening of clonal selections. This knowledge must be taken into account in the planning of future breeding projects. During this project a total of 38 984 seedlings were screened and four selections were made. For both the nursery and the producer, knowledge of the clonal ability of a potential new rootstock is important from a financial point of view. / Dissertation (MSc (Horticultural Science))--University of Pretoria, 2006. / Plant Production and Soil Science / unrestricted

As the causative agent of black shank, Phytophthora nicotianae is a serious threat to tobacco cultivation in South Africa. Research presented in this dissertation describes pathogenicity studies and control measures for P. nicotianae on tobacco. Special attention is given to the population structure of P. nicotianae in South Africa. The implications of these genetic studies in breeding and selection programs against P. nicotianae were also evaluated. The first chapter of this dissertation represents a literature review on black shank and available control measures for P. nicotianae on tobacco. The mechanisms of pathogenicity and the life cycle of P. nicotianae are also treated in detail. Special reference is made to the maintenance of genetic diversity in Phytophthora species and particularly P. nicotianae. This literature review also highlights the fact that very few studies have been conducted to determine the genetic structure of P. nicotianae populations. The success of South African breeding programs for tobacco cultivars with P. nicotianae resistance is to some degree dependent on the selection of isolates with high levels of aggressiveness. The research presented in chapter two provides information on cultivar resistance and selection of P. nicotianae isolates for future breeding programs. Significant differences in levels of aggressiveness were found between P. nicotianae isolates. Furthermore, race 0 and 1 of P. nicotianae occurred in most of the tobacco growing regions in South Africa. Selected Race 0 and 1 isolates were thus used to evaluate black shank resistance of 11 commercially planted tobacco cultivars. Commercially planted cultivars differed significantly in their resistance to race 0 and 1. Cultivars LK33/60 and OD1 were highly resistant to race 0 but susceptible to race 1 while cultivars Vuma/3/46 and LK3/46 were highly resistant to both race 0 and 1. Chapter three reports on the use of metalaxyl treatments combined with resistance in tobacco cultivars for control of P. nicotianae. One hundred and thirty two isolates of P. nicotianae were screened for sensitivity to metalaxyl. P. nicotianae isolates from most tobacco farms were metalaxyl sensitive. The results further indicated that the use of metalaxyl in combination with moderately resistant cultivars effectively reduced black shank in the field. The outcome of this study provided useful information for the implementation of an economically viable combination of disease resistance and metalaxyl as the basis for a P. nicotianae management program in South Africa. Chapter four of this dissertation deals with the development of a rapid seedling-' based screening technique to assay tobacco for resistance to P. nicotianae. This technique was validated by comparing it to a stem inoculation technique commonly used on adult plants. A strong positive correlation was found between results of the seedling assay and adult plant trials for all isolates and cultivars tested. P. nicotianae isolates could also be characterized as race 0 or I using both stem inoculation and the rapid seedling assay. The ability to screen large numbers of tobacco plants rapidly at the seedling stage allows for the testing of large germplasm resources in a systematic manner and under standard conditions. This may help in the timely development and release of more black shank resistant cultivars. In chapter five, a population study on P. nicotianae in South Africa is presented. One hundred and five P. nicotianae isolates were collected from the Northern Highveld and Lowveld regions, as well as from both citrus and tobacco hosts in South Africa. Levels of phenotypic diversity were determined in populations of P. nicotianae using RAPD markers. Among the 105 P. nicotianae isolates analysed 79 different RAPD phenotypes were found, where 35 of the isolates were found to be clonal. The high number of RAPD phenotypes (79) in relation to the sample size (105), the presence of both the Al and A2 mating type and high levels of phenotypic diversity in the P. nicotianae population indicate a sexually outcrossing P. nicotianae population in South Africa. This sexual outcrossing may mean that P. nicotianae is likely to remain a constant threat to tobacco and citrus cultivation, since new genotypes with the potential to overcome resistance genes in commercial cultivars are likely to emerge. All chapters of this dissertation deal with some aspects of black shank control and breeding for resistance to P. nicotianae. This dissertation provides new knowledge on variation in levels of aggressiveness, race distribution and the development of metalaxyl resistance in the South African P. nicotianae populations. This also represents the first study on the genetic diversity of P. nicotianae populations in South Africa. The results presented here not only show the possible occurrence of sexual reproduction, but also indicate the presence of clones and discreet phenotypic groups of P. nicotianae. This information will be applied in future tobacco breeding programs to select breeding lines with resistance against a number of specific P. nicotianae races and phenotypic groups. / Thesis (DPhil)--University of Pretoria, 2006. / Microbiology and Plant Pathology / Unrestricted

USAID / Se identifica la expresión diferencial de genes efectores tipo RXLR en dos cepas aisladas del centro de los andes peruanos de P. infestans EC-1 mediante secuenciamiento del transcriptoma de la interacción papa-P.infestans de los primeros días después de la infección, siendo confirmada por qRT-PCR. Los genes efectores fueron silenciados en una cepa para Avr-vnt1 en POX109 y para el homólogo Avh9.1 en POX067, pero expresados en la otra. Además, los resultados de transcriptoma fueron comparados con tres cepas adicionales del linaje EC-1. En el análisis de SNPs de Avr-blb1, Avr-blb2 y Avr-vnt1, la variabilidad alélica no tuvo predominancia frente a la variabilidad de expresión de genes. Asimismo, debido al silenciamiento génico de Avr-vnt1 se evaluó la expresión de estos en plantas transgénicas [Rpi-vnt1.1] a fin de encontrar si la resistencia transgénica era funcional. Encontrando que en ambas cepas en todos los eventos y en el control susceptible Yungay se expresan, a diferencia del resultado anterior. El descubrimiento de efectores silenciados en las poblaciones del patógeno pueden guiar al uso de genes R específicos en los programas de mejoramiento genético. Pudiendo el gen Rpi-vnt1 no ser recomendado. / Tesis