Crown rust, caused by the fungus Puccinia coronata, is a severe disease negatively impacting seed quality and yield in oat (Avena sativa). Host genetic resistance is the primary means for controlling this disease. The most extensive oat map contains nearly 2,500 genetic markers, many of which are restriction and amplified fragment length polymorphic (RFLP or AFLP) markers. However, the use of more abundant single nucleotide polymorphic (SNP) markers combined with diversity arrays technology (DArT) would be more advantageous for marker assisted breeding (MAB) and genome wide selection (GWS) applications due to the availability of high density genotyping technologies. The purpose of using this technology is to improve the competitiveness of oat by producing varieties with durable resistance to crown rust and desirable traits that will benefit oat growers in the U.S. Panels of winter and spring oat were evaluated for resistance to crown rust in four field environments in Texas, Louisiana, Minnesota, and North Dakota during a two-year study in 2010 and 2011. Plants representing 702 elite lines of oat were phenotyped for crown rust resistance and found to have highly diverse responses. The winter oat lines demonstrated the best crown rust resistance and are expected to yield the most QTL to be used in developing durable crown rust resistance. Heritability of crown rust resistance in this study ranged from 0.88 to 0.90 in spring and winter oats, respectively. Crown rust measurements were also found to be repeatable. Repeatability ranged from 0.56 to 0.88 at Castroville, TX in 2011 and 2010, respectively in spring oats and from 0.79 at St. Paul, MN in 2011 to 0.96 at Castroville, TX in 2010 in winter oats. Oat lines contributed by states along the Puccinia pathway in Texas, Louisiana, Minnesota, North Dakota, and Wisconsin on average exhibited the best crown rust resistance as compared to other areas in the country where spring and winter oat are grown. GGE biplot analysis indicated that Castroville, TX was the most representative and most ideal testing location. The above results are expected to increase knowledge of the genetic diversity of the oat germplasm, yield comprehensive genotyping and phenotyping information for North American oat breeding programs, and to promote further use of GWS and MAB for key traits regarding disease resistance in oat. Future work is to conclude the association mapping process by completing genotypic analysis.

The aim of this work is the accurate diagnosis of the Puccinia species of the selected turf grass species under field and laboratory conditions and evaluate influences on the intensity of occurrence. The laboratory tests will be recommended for the best stage of plants to artificial infection. The experiment was carried out in collaboration with SŠ Větrov. In the two-year follow-up (in 2011 and 2012) were at particular times of samples of infected plants, which were then evaluated microscopically. And at the same time experimental work was carried out in laboratory conditions (artificial infection) in plant material of the same origin and assessed degree assault with a precise diagnosis of the type of pathogen. Between 2011 and 2012, was higher occurence stem rust than crown rust.. From a total of 514 tests was stem rust in 379 cases, while crown rust in only 40 cases. In 95 samples was observed incidence of both simultaneously rust and mildew spores remaining plants were discovered. Greenhouse resistance tests were performed on younger plants (3 sheets) and older (6 sheets) in 16 variants, of which 4 were control. Infected leaves were spray uredispor with talc and injected with an aqueous suspension followed by looking drops and were then sealed under insulators at 23 ° C for 24 - 48 hours.

The aim of the thesis is evaluation of occurence of rusts on the selected types of grass, their specific diagnosis and valorization of their influence on the important agricultural qualities. The experiment was carried out in a cooperation with SS Větrov. In a two-year period (in 2009 and 2010) the samples of infested plants, which were subsequently evaluated microscopically, were taken. The ascertained results were processed through the Kruskal-Wallis test and the Independent Group T-Test. In 2009 and 2010 the occurrence of stem rust on the tested types of grass was significantly higher than occurence of crown rust. From the total number of 513 tests stem rust was found in 353 cases, whereas crown rust was found only in 103 cases. In 27 samples the occurrence of both types of rusts was found and there was no occurence found on the rest of the plants. The results show that in our climatic conditions stem rust becomes the most frequent pathogen which causes rustiness of grass. In 2010 the results of turf experiments proved that the selection of genotype of perenne reygrass with a lower infestation by stem rust had a positive impact on health conditions and better appearance of the offspring of the selected plans. Effectivenes of selection was confirmed. Among the tested materials there were chosen very predisposed genotypes which could be useful as the infectious material in the planned greenhouse tests for the resistance against rusts. Currently, we have also been abble to recognize the genotypes which are relatively resistant against particular rusts. If their resistance is confirmed in the following period, they might be used as donors of resistance in xenogamy and as immune standards in tests. The introduced results from the first stage of cooperation are only preliminary and it will be necessary to verify them at the following stages.

Made available in DSpace on 2015-03-26T13:37:55Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1312640 bytes, checksum: f59984d596fc23251bdcc4b712cc574a (MD5) Previous issue date: 2014-07-29 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Rust fungi are obligate parasites that secrete diverse proteins during their interaction with host plants. These proteins promote parasitism by manipulating the host metabolism and interfering with its defense responses. Effector proteins can be recognized by proteins coded by plant resistance genes, in which case they are called avirulence proteins. Six loci containing genes that confer varying degrees of resistance to P. pachyrhizi (Rpp) have been reported, and several genes encoding proteins secreted by the fungus have been identified. However, effector proteins (Avr proteins) recognized by soybean Rpp proteins have not yet been identified. Thus, the objective of this study was to identify proteins secreted by P. pachyrhizi (PPUFV02) able to activate or suppress defense responses in soybean, through the functional analysis of ten candidate effector genes using the TTSS of P. savastanoi pv. glycinea 1462 (EDV system). Among the ten effectors tested in 11 soybean genotypes resistant to the mono-pustule isolate PPUFV02 of P. pachyrhizi, source of candidate genes, PHPA_RSP_71 stood out for contributing to greater disease severity in genotype PI 594538- A and lower in genotype PI 594754. The PHPA_RSP_23 effector was also interesting for additional functional studies on soybean and tobacco, since the results indicated a possible activation of defense response in soybean, in general (all genotypes), and a possible suppression of cell death in tobacco. The effector activity of these candidate genes was not confirmed by analysis of bacterial growth in soybean and tobacco (non-host plant). On the other hand, it is possible that these effects are due to an interference of the effector with the function of the TTSS, making it necessary to conduct protein secretion assays to confirm or reject these hypotheses. Overall, the genotype Williams 82 showed resistance to Psg 1462 and can be used as negative control for disease in functional analysis in soybean. It was not possible to confirm N. benthamiana as an appropriate plant for functional studies of induction/suppression of HR when using Psg 1462, since it is possible that the bacterial isolate is pathogenic to this species. The pEDV6 vector in general was stable in the Psg1462(pEDV6::PHPA_RSP) clones and did not alter the ability of the bacteria to multiply in planta, confirming the EDV as a good system for functional analysis of P. pachyrhizi candidate effector genes in soybean using TTSS of Psg 1462. The inoculation method by dipping was not suitable for functional analysis in soybean because, in general, showed highly variable results. The inoculation method by vacuum was the best suited for this purpose, since it introduced greater uniformity in the severity of symptoms observed. / Os fungos causadores de ferrugens são parasitas obrigatórios que secretam diversas proteínas durante sua interação com plantas hospedeiras. Essas proteínas promovem o parasitismo por meio da manipulação do metabolismo do hospedeiro e interferência em suas respostas de defesa. As proteínas efetoras podem ser reconhecidas pelas proteínas codificadas pelos genes de resistência da planta, sendo, neste caso, denominadas proteínas de avirulência. Seis locos contendo genes que proporcionam diferentes graus de resistência à P. pachyrhizi (Rpp) já foram relatados e vários genes que codificam proteínas secretadas por esse fungo já foram identificados. No entanto, ainda não foram identificadas as proteínas efetoras (proteínas Avr) reconhecidas pelas proteínas Rpp de soja. Desse modo o objetivo deste trabalho foi identificar proteínas secretadas por P. pachyrhizi (PPUFV02), capazes de ativar ou suprimir respostas de defesa em soja, através de uma análise funcional de 10 genes candidatos a efetores, utilizando o SST3 de P. savastanoi pv. glycinea 1462 (sistema EDV). Dos dez efetores testados nos 11 genótipos de soja resistentes ao isolado monopostular de P. pachyrhizi PPUFV02, fonte dos genes candidatos, PHPA_RSP_71 se destacou por contribuir para uma maior severidade da doença no genótipo PI 594538-A e menor no genótipo PI 594754. O efetor PHPA_RSP_23 também se mostrou interessante para novos estudos funcionais em soja e tabaco, uma vez que os resultados indicaram possível ativação de resposta de defesa em soja de forma geral (em todos os genótipos) e possível supressão de morte celular em tabaco. A atividade efetora desses genes candidatos não foi confirmada pela análise de crescimento bacteriano em soja e em tabaco (planta não hospedeira). Por outro lado, há uma possibilidade de esses efeitos serem devido a uma interferência do efetor com a funcionalidade do SST3, necessitando-se de estudos de secreção da proteína para confirmar ou refutar estas hipóteses. De forma geral o genótipo Williams 82 se mostrou resistente à Psg 1462, podendo ser utilizado como controle negativo para doença nas análises funcionais. Não foi possível confirmar N. benthamiana como uma planta adequada para estudos funcionais de indução/supressão de HR ao se utilizar Psg 1462, uma vez que é possível que este isolado bacteriano seja patogênico a esta espécie. O vetor pEDV6 de forma geral apresentou estabilidade nos clones Psg1462(pEDV6::PHPA_RSP) e não alterou a capacidade de multiplicação da bactéria in planta, confirmando o EDV como um bom sistema para análise funcional de genes de P. pachyrhizi candidatos a efetores em soja, utilizando o SST3 de Psg 1462. O método de inoculação por mergulho não se mostrou adequado para as análises funcionais em soja, uma vez que apresentou de forma geral alta variabilidade nos resultados, sendo o método de inoculação a vácuo mais indicado para este fim por ter apresentado maior uniformidade na severidade dos sintomas observados.