A maioria das plantas consegue defender-se contra infecções fúngicas por meios naturais, que podem ser induzidos por uma série de elicitores bióticos ou abióticos. O silício (Si) solúvel tem mostrado que é capaz de induzir resistência em várias espécies de plantas contra inúmeros patógenos. Neste trabalho foi proposta a avaliação dos compostos contento Si na redução dos sintomas da ferrugem causada pelo fungo biotrófico Hemileia vastatrix em plantas de cafeeiros suscetíveis bem como avaliar os parâmetros bioquímicos envolvidos nos processos de resistência. Primeiramente, foram estudadas duas fontes de Si (silicato de Ca/Mg e silicato de potássio) em cinco doses para padronizar uma dose/fonte para os experimentos futuros. Foi possível observar que as plantas de cafeeiros não tiveram diferença significativa nos parâmetros de desenvolvimento como altura das plantas, área foliar, número de folhas para nenhuma das fontes e doses analisadas. Porém quando se tratava da contagem do número de lesões por cm2, a fonte silicato de potássio na dose de 5mM conseguiu suprimir em até 60% o desenvolvimento das lesões causadas pelo fungo. Com a fonte e dose estabelecidas, o segundo passo do trabalho foi avaliar quais os processos bioquímicos envolvidos na resistência conferida pelo Si em plantas de cafeeiro. Foram analisadas as atividades das enzimas relacionadas ao estresse oxidativo (peroxidases guaiacol e ascorbato, catalases, e superóxido desmutases) e relacionadas à defesa (glucanase, quitinase e PAL). As folhas foram coletadas para a obtenção do extrato protéico em diferentes intervalos de tempo após a inoculação com o fungo: as 24, 48, 72 e 96 horas após a inoculação e como controle foi utilizado o tempo zero (sem inoculação). Nesta fase foi possível observar que a atividade das enzimas CAT, SOD, APX foi maior em plantas tratadas indicando que o Si parece estimular uma resposta mais rápida ao estresse oxidativo. O mesmo ocorreu com as enzimas relacionadas à defesa. A partir destes resultados podemos afirmar que o Si estimula uma resposta de defesa mais rápida em plantas de café suscetíveis à ferrugem quando inoculadas com o fungo patogênico / Most plants can defend themselves against fungal infections by natural means, which can be induced by a number of biotic and abiotic elicitors. Soluble silicon (Si) has been shown to induce resistance in a number of plant species against several pathogens. The objective of this work was to assess the influence of silicon application of symptoms caused by the biotrophic fungus Hemileia vastatrix in susceptible coffee plants and evaluate the biochemistry process involved in defense mechanism. First of all, were studied two Si sources (Ca/Mg silicate and potassium silicate) in five doses to standardize dose/source for the future experiments. The statistical analysis showed that no difference was observed in relation to plant height, leaf area and number of leaves. However, the number of lesions reduced 60% in the higher silicon dose when compared to the number of lesions in control plants. Infected plants were found to have a linear decrease of lesions with the increase of silicate concentration. The lowest number of lesions per leaf area was observed in plants that received 5 M of Si from potassium silicate. This result indicates the use of silicon as an alternative for an ecological management system for coffee disease protection. The second step of this work was to evaluate the biochemistry process involved in resistance induced by Si in susceptible coffee plants. The activity of the enzymes related to oxidative stress (peroxidases, catalase, and superoxide desmutase) and related to defense (glucanase, chitinase, PAL) was evaluated. The leaves were collected in different time intervals after the fungus inoculation (24, 48, 72 e 96 hours after the fungus inoculation) in order to obtain the protein extract. It was possible to observe that the activity of CAT, SOD and APX was higher in Si-treated coffee plants suggesting that Si could stimulate a faster response to oxidative stress. The same situation occurred to the defense related enzymes. By the results obtained here it is possible to verify that Si can stimulate a faster defense response in susceptible coffee plants when inoculated with coffee leaf rust fungus

Three hundred ninety one (391) spring wheat introductions from Asian, Middle Eastern and Mediteranean areas were screened for resistance to races C17, C20, C25, C49, C53 and C57 of Puccinia qraminis tritici; races 1, 5, 9, 15 and bulks 1, 4 and l0 of Puccinia recondita; races T2, T10, T13 and T39 of Ustilago tritici and to 6 isolates of Drechslera tritici-repentis prevalent in western Canada. Of the 34 introductions resistant to P. graminis tritici, 15 were genetically studied using F2 segregation data derived from the progeny of the crosses involving resistant introductions, their corresponding near isogenic lines and stem rust universal suscepts. Eleven of these were found to carry single Sr genes for resistance, notably, Sr30, Sr13 and Sr15. Of the 70 introductions resistant to P. recondita, 28 were studied and 9 were found to carry known Lr genes for resistance, notably Lr10 and the genes present in RL6057 and RL6061. Twenty two introductions are believed to carry either 1 or 2 unidentified dominant, recessive, partially dominant and/or complementary genes for resistance to either stem or leaf rust. Five introductions were immune and 6 highly resistant to the 4 races of U. tritici. Sixty-nine introductions were resistant to D. tritici-repentis . These were arbitrarily classified into 10 'phenotypic classes' according to their reactions to the 6 isolates used.

Wheat is infected by leaf rust disease (Puccinia triticina Eriks.) almost everywhere it is cultivated. The dynamic nature of P. triticina populations affects the effective life span of genes conferring leaf rust resistance (Lr genes). Genetic diversity and combinations of Lr genes should be used to achieve durable resistance. Toropi, a Brazilian wheat cultivar, has maintained leaf rust resistance since its release in 1965, suggesting that it is a good candidate for durable resistance. Two recessive complementary adult plant genes were previously described in Toropi. The objective of this study was to characterize and map the sources of resistance present in Toropi. Double haploid (DH) populations developed by crossing the susceptible leaf rust cultivar Thatcher with Toropi were analysed in Canada (Glenlea – 2010 and Portage La Prairie – 2011), New Zealand (Lincoln – 2010 and 2011) and in Brazil (Parana – 2011), and in greenhouse. The leaf rust reactions indicated the presence of at least four leaf rust resistance genes in Toropi: one seedling gene and three adult plant genes. The seedling resistance gene Trp-Se, responsible for immune response in New Zealand, was mapped on chromosome 3D. QTL analyses identified a QTL associated with leaf rust resistance (QLr.crc-5AL.1) on chromosome 5AL, which overlapped with a QTL for stripe rust (QStr.crc-5AL.1) in the same population. This gene, designated Trp1, is believed to be one of the two adult plant complementary partial resistance genes. The position of the Trp-2 is not confirmed yet. One minor race specific adult plant gene, temporarily designated Trp-3, was mapped on 4BL chromosome. The Lr genes in Toropi confer minor effects against leaf rust, except for Trp-Se, which conditioned immunity in New Zealand. However, when the Toropi Lr genes were combined an almost immune response resulted. Toropi had a very good leaf rust resistance in South and North America, and in New Zealand. The molecular markers identified during this project could facilitate the incorporation of the Toropi genes in new cultivars, helping to achieve more diverse and durable wheat.

Three hundred ninety one (391) spring wheat introductions from Asian, Middle Eastern and Mediteranean areas were screened for resistance to races C17, C20, C25, C49, C53 and C57 of Puccinia qraminis tritici; races 1, 5, 9, 15 and bulks 1, 4 and l0 of Puccinia recondita; races T2, T10, T13 and T39 of Ustilago tritici and to 6 isolates of Drechslera tritici-repentis prevalent in western Canada. Of the 34 introductions resistant to P. graminis tritici, 15 were genetically studied using F2 segregation data derived from the progeny of the crosses involving resistant introductions, their corresponding near isogenic lines and stem rust universal suscepts. Eleven of these were found to carry single Sr genes for resistance, notably, Sr30, Sr13 and Sr15. Of the 70 introductions resistant to P. recondita, 28 were studied and 9 were found to carry known Lr genes for resistance, notably Lr10 and the genes present in RL6057 and RL6061. Twenty two introductions are believed to carry either 1 or 2 unidentified dominant, recessive, partially dominant and/or complementary genes for resistance to either stem or leaf rust. Five introductions were immune and 6 highly resistant to the 4 races of U. tritici. Sixty-nine introductions were resistant to D. tritici-repentis . These were arbitrarily classified into 10 'phenotypic classes' according to their reactions to the 6 isolates used.

Wheat is infected by leaf rust disease (Puccinia triticina Eriks.) almost everywhere it is cultivated. The dynamic nature of P. triticina populations affects the effective life span of genes conferring leaf rust resistance (Lr genes). Genetic diversity and combinations of Lr genes should be used to achieve durable resistance. Toropi, a Brazilian wheat cultivar, has maintained leaf rust resistance since its release in 1965, suggesting that it is a good candidate for durable resistance. Two recessive complementary adult plant genes were previously described in Toropi. The objective of this study was to characterize and map the sources of resistance present in Toropi. Double haploid (DH) populations developed by crossing the susceptible leaf rust cultivar Thatcher with Toropi were analysed in Canada (Glenlea – 2010 and Portage La Prairie – 2011), New Zealand (Lincoln – 2010 and 2011) and in Brazil (Parana – 2011), and in greenhouse. The leaf rust reactions indicated the presence of at least four leaf rust resistance genes in Toropi: one seedling gene and three adult plant genes. The seedling resistance gene Trp-Se, responsible for immune response in New Zealand, was mapped on chromosome 3D. QTL analyses identified a QTL associated with leaf rust resistance (QLr.crc-5AL.1) on chromosome 5AL, which overlapped with a QTL for stripe rust (QStr.crc-5AL.1) in the same population. This gene, designated Trp1, is believed to be one of the two adult plant complementary partial resistance genes. The position of the Trp-2 is not confirmed yet. One minor race specific adult plant gene, temporarily designated Trp-3, was mapped on 4BL chromosome. The Lr genes in Toropi confer minor effects against leaf rust, except for Trp-Se, which conditioned immunity in New Zealand. However, when the Toropi Lr genes were combined an almost immune response resulted. Toropi had a very good leaf rust resistance in South and North America, and in New Zealand. The molecular markers identified during this project could facilitate the incorporation of the Toropi genes in new cultivars, helping to achieve more diverse and durable wheat.

A maioria das plantas consegue defender-se contra infecções fúngicas por meios naturais, que podem ser induzidos por uma série de elicitores bióticos ou abióticos. O silício (Si) solúvel tem mostrado que é capaz de induzir resistência em várias espécies de plantas contra inúmeros patógenos. Neste trabalho foi proposta a avaliação dos compostos contento Si na redução dos sintomas da ferrugem causada pelo fungo biotrófico Hemileia vastatrix em plantas de cafeeiros suscetíveis bem como avaliar os parâmetros bioquímicos envolvidos nos processos de resistência. Primeiramente, foram estudadas duas fontes de Si (silicato de Ca/Mg e silicato de potássio) em cinco doses para padronizar uma dose/fonte para os experimentos futuros. Foi possível observar que as plantas de cafeeiros não tiveram diferença significativa nos parâmetros de desenvolvimento como altura das plantas, área foliar, número de folhas para nenhuma das fontes e doses analisadas. Porém quando se tratava da contagem do número de lesões por cm2, a fonte silicato de potássio na dose de 5mM conseguiu suprimir em até 60% o desenvolvimento das lesões causadas pelo fungo. Com a fonte e dose estabelecidas, o segundo passo do trabalho foi avaliar quais os processos bioquímicos envolvidos na resistência conferida pelo Si em plantas de cafeeiro. Foram analisadas as atividades das enzimas relacionadas ao estresse oxidativo (peroxidases guaiacol e ascorbato, catalases, e superóxido desmutases) e relacionadas à defesa (glucanase, quitinase e PAL). As folhas foram coletadas para a obtenção do extrato protéico em diferentes intervalos de tempo após a inoculação com o fungo: as 24, 48, 72 e 96 horas após a inoculação e como controle foi utilizado o tempo zero (sem inoculação). Nesta fase foi possível observar que a atividade das enzimas CAT, SOD, APX foi maior em plantas tratadas indicando que o Si parece estimular uma resposta mais rápida ao estresse oxidativo. O mesmo ocorreu com as enzimas relacionadas à defesa. A partir destes resultados podemos afirmar que o Si estimula uma resposta de defesa mais rápida em plantas de café suscetíveis à ferrugem quando inoculadas com o fungo patogênico / Most plants can defend themselves against fungal infections by natural means, which can be induced by a number of biotic and abiotic elicitors. Soluble silicon (Si) has been shown to induce resistance in a number of plant species against several pathogens. The objective of this work was to assess the influence of silicon application of symptoms caused by the biotrophic fungus Hemileia vastatrix in susceptible coffee plants and evaluate the biochemistry process involved in defense mechanism. First of all, were studied two Si sources (Ca/Mg silicate and potassium silicate) in five doses to standardize dose/source for the future experiments. The statistical analysis showed that no difference was observed in relation to plant height, leaf area and number of leaves. However, the number of lesions reduced 60% in the higher silicon dose when compared to the number of lesions in control plants. Infected plants were found to have a linear decrease of lesions with the increase of silicate concentration. The lowest number of lesions per leaf area was observed in plants that received 5 M of Si from potassium silicate. This result indicates the use of silicon as an alternative for an ecological management system for coffee disease protection. The second step of this work was to evaluate the biochemistry process involved in resistance induced by Si in susceptible coffee plants. The activity of the enzymes related to oxidative stress (peroxidases, catalase, and superoxide desmutase) and related to defense (glucanase, chitinase, PAL) was evaluated. The leaves were collected in different time intervals after the fungus inoculation (24, 48, 72 e 96 hours after the fungus inoculation) in order to obtain the protein extract. It was possible to observe that the activity of CAT, SOD and APX was higher in Si-treated coffee plants suggesting that Si could stimulate a faster response to oxidative stress. The same situation occurred to the defense related enzymes. By the results obtained here it is possible to verify that Si can stimulate a faster defense response in susceptible coffee plants when inoculated with coffee leaf rust fungus

Master of Science / Department of Agronomy / John P. Fellers / Allan K. Fritz / Water availability is commonly the most limiting factor to crop production, especially in drought prone areas like the Midwest. This study was conducted to map quantitative trait loci (QTL) involved in drought tolerance in wheat (Triticum aestivum L.) to enable their use for marker assisted selection (MAS) in breeding. A population of 122 F[subscript]7 derived recombinant inbred lines from a cross between Dharwar Dry and Sitta, spring wheat lines with contrasting drought tolerances, was analyzed using the amplified fragment length polymorphism (AFLP) technique and Diversity Array Technology (DArT) markers to create a QTL map. Of the 256 AFLP primer combinations evaluated, 151 were found to be polymorphic between the parents and were used to screen the population. A linkage map of 48 groups was created from the combined DArT markers, AFLP data, and SSR markers. This was used to create a QTL map which identified QTL in 24 of these groups. Using these markers for MAS in a breeding program could overcome the difficulties of selecting for drought tolerance. Another serious limitation to wheat production is leaf rust caused by the pathogen Puccinia triticina. Leaf rust causes between 1% and 20% yield loss on average and tends to be the worst in years with high yield potential. PI 289824 contains a single, dominant gene for seedling resistance mapping to chromosome 5BS and thought to be different from Lr52. An F[subscript]2 mapping population from a cross between PI 289824 and Jagger was used to try to identify markers very closely linked to the gene and therefore useful for MAS. The population presented some mapping challenges, but with the use of SSR and EST-STS markers, the gene was flanked. However, the markers were at too a great distance to be useful for mapping.

Wheat is the most widely grown crop in the world and as such, is an essential source of energy and nutrition. The challenges that breeders presently face is to increase production to feed the rising population of the world, while also accounting for climate change, pollution, water and environmental stresses. As genetic uniformity of modern cultivars has increased vulnerability to pests and diseases, the wild relatives of wheat offer a rich source of genetic diversity and stress tolerance traits, that can be harnessed and transferred in to modern wheat. In this study, we used array-based genotyping to explore genetic diversity in 385 domesticated and non-domesticated lines of wheat and their wild relatives. Genetic characterization using the wheat 90K array, and subsequent filtering and validation mapped 9,570 single nucleotide polymorphic markers onto the wheat reference genome. Phylogenetic analyses illustrated four major clades, clearly separating the wild species from the domesticated, and the ancient Triticum turgidum species from modern T. turgidum cultivars. Using this diverse germplasm, a genome-wide association study (GWAS) was performed for leaf rust, the most widespread rust disease of wheat. Identification of novel sources of resistance is necessary to maintain disease resistance and stay ahead in the plant-pathogen evolutionary arms race. GWAS was conducted using eight statistical models for infection types against six leaf rust isolates and leaf rust severity rated in field trials for 3-4 years at 2-3 locations in Canada. Functional annotation of genes containing significant quantitative trait nucleotides (QTNs) identified 96 disease-related nucleotide associated with leaf rust resistance. A total of 21 QTNs were in haplotype blocks or within flanking markers of at least 16 known leaf rust (Lr) resistance genes. The remaining significant QTNs were considered loci that putatively harbor new Lr resistance genes. Future efforts to validate these loci will help understand their role in disease resistance and promote their utility for marker-assisted selection in pre-breeding.

Doctor of Philosophy / Department of Agronomy / Allan K. Fritz / In this research, a recombinant inbred line (RIL) population derived from cross between a leaf rust- and stripe rust-susceptible spring wheat ‘Avocet S’ and a slow leaf- and stripe-rusting resistant spring wheat ‘Amadina’ was used to postulate and map leaf rust seedling resistance genes, identify quantitative trait loci (QTL) for slow-rusting resistance against leaf and stripe rust, and study slow leaf-rusting components, latent period and infection frequency. Two known Lr genes (Lr23, and Lr26) were identified to be present in ‘Amadina’ through gene postulation, pedigree, cytogenetic, and polymerase chain reaction analyses. One unknown gene associated with seedling resistance was also mapped on chromosome 1BL. In greenhouse experiment, it was estimated that at least five genes conditioning final disease severity (FS) and latent period (LP), and four genes conditioning infection frequency (IF), segregated in the population. Correlations between LP and FS, and LP and IF were moderately negative, and that between IF and FS was moderately positive, indicating inter-dependence of the traits. Two QTL on chromosomes 1BL and 6BL were associated with LP and FS, and three QTL on chromosomes 1BL, 6BL and 2DS were associated with IF. Segregation of the RIL population in field experiment indicated that there were at least four and three adult plant resistance (APR) genes involved in resistance for leaf and stripe rust. Six QTL on chromosomes 3AL, 4AL, 1BL, 5BL, and 7BL were associated with APR for leaf rust, and seven QTL on chromosome 4AL, 5AL, 1BL, 2BL, 4BL, 5BL, 2DL, and 4D were associated with APR for stripe rust. Our results indicated that the major portion of genetic variability for slow-rusting resistance was additive gene action, and, to some extent, epistasis. In this research, we also explored the utility of remote sensing and geographic information systems (GIS) and analytical operations to discriminate leaf rust pustules from other parts of leaf and to accurately determine pustule size in ‘Amadina’ and ‘Avocet S’.

Call number: LD2668 .T4 1979 C66 / Master of Science