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Made available in DSpace on 2017-07-10T17:37:37Z (GMT). No. of bitstreams: 1 Josiel_Pereira_Nascimento.pdf: 909698 bytes, checksum: 3cb3ef0ef8bdf27d1c63a9fb56af8240 (MD5) Previous issue date: 2009-08-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The non tillage planting and the cultivation of safrinha (short season) corn are factors that may provide alterations in the microclimate and in the biology of agro ecosystem, increasing the populations of fungus like Gibberella zeae, which is the etiologic agent of one of the maize ear rots. This work had the objective to identify inoculation methods that can be useful in the election and in the characterization of genotypes tolerant to G. zeae. The experiment was conducted in Toledo/PR during the 2008 safrinha season in a randomized complete block design with five replications, on a factorial scheme where hybrids were the first factor (2B710; 30F45; DKB 214; 30F98; DKB 390; GAC 9020; 30F80; 30R50 and 30K73) and the inoculation methods were the second factor. The hybrids had been grouped as susceptible (2B710; 30F45; DKB 214; 30K73), tolerant (DKB 390; GAC 9020; 30F80; 30R50) and moderately susceptible to giberela (30F98). The 12 evaluated methods consisted of simulating the fungus natural infection on the silk or on the damage caused by insects in the lateral of the ears, and also varying by the absence of covering on the ear (humid chamber), by covering the ear for 24 hours and by covering the ear for 48. The methods with inoculation of 2 mL in the lateral of the ear without covering (3), with covering for 24 hours (7) and with covering for 48 hours (11) had shown efficiency in the characterization of tolerant and susceptible hybrids and could be used on a large scale in the identification and selection of maize varieties with tolerance to ear rot caused by G. zeae. As a result, the seed industry can provide superior materials and the farmers will reduce their losses due to kernel quality. Moreover, the whole society will consume better maize products, with less mycotoxins / O plantio direto e o cultivo de milho safrinha são fatores que podem proporcionar alterações no microclima e na biologia do agroecossistema, aumentando as populações de fungos como Gibberella zeae, agente etiológico da doença denominada giberela ou podridão-vermelha-da-espiga. Esse trabalho teve como objetivo identificar métodos de inoculação que auxiliassem na seleção e na caracterização de genótipos tolerantes à G. zeae. O experimento foi conduzido no município de Toledo/PR durante a safrinha de 2008. O delineamento experimental foi em blocos ao acaso com cinco repetições, em esquema fatorial tendo como primeiro fator os híbridos (2B710; 30F45; DKB 214; 30F98; DKB 390; AG 9020; 30F80; 30R50 e 30K73) e como segundo fator os métodos de inoculação. Os híbridos foram agrupados como suscetíveis (2B710; 30F45; DKB 214; 30K73), tolerantes (DKB 390; AG 9020; 30F80; 30R50) e moderadamente suscetível a giberela (30F98). Os 12 métodos avaliados consistiam em simular a infecção natural do fungo pelo estigma ou por danos causados por insetos nas laterais das espigas, variando ainda em ausência de cobertura (câmara úmida), cobertura por 24 horas e por 48 horas. Os métodos com inoculação de 2 mL na lateral da espiga sem cobertura (3), com cobertura por 24 horas (7) e com cobertura por 48 horas (11) se mostraram eficientes na caracterização de híbridos tolerantes e suscetíveis e poderão ser empregados em larga escala na identificação e seleção de variedades de milho tolerantes à doença. Mais do que isso, a sociedade poderá consumir produtos de milho com qualidade superior, com menos micotoxinas

Made available in DSpace on 2016-12-06T17:42:07Z (GMT). No. of bitstreams: 1 PGPV14DA016.pdf: 1791236 bytes, checksum: 435d76b4d49498dd4bdc194423dac320 (MD5) Previous issue date: 2014-10-17 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The gibberella ear rot caused by fungus Fusarium graminearum, causes damage in productivity and quality of maize kernel being disease of difficult to control due to the unavailability of resistant hybrids and low efficacy of cultural practices. Three studies were conducted, the aim of the first was to compare four methods of inoculation (injection silkchannel, sprinkling on silk, kernel- wounding and deposition in the ear leaf sheath) and a control treatment in four hybrids (P30R50H, P30B30H, P3989, P32R48H) based on disease severity and kernel yield; the second study determined the reaction of 20 hybrids to gibberella ear rot by quantifying the incidence of the disease, rot kernels and F. graminearum in the kernels and their correlations; and the third study characterized the genetic resistance of hybrids to gibberella ear rot through preparation of a reaction scale involving classes and disease intensity and their correlation with kernel yield and inoculation methods. The experiments of first study were conducted in 2011/12 harvest in Cruz Alta, RS and Lages. The experiments of the second and third studies were conducted in two environments in the season 2012/13 in Cruz Alta. The design of the experiments was a randomized complete blocks design with treatments arranged in a factorial with four replications. In the first study there was no difference in disease severity between hybrids and inoculation methods. The hybrid P32R48H presented higher disease severity, distinguishing from the others. Inoculations in silk-channel and wounding the kernels provided higher disease severity and lower kernel yield. In the second study, the method of the wounding the kernels provided increases above 40% in the incidence of gibberella ear rot, rot kernels and occurrence of F. graminearum in the grain when compared to the injection silk-channel method. There was an effect of hybrids and the values were 37.2% and 95.6% for the incident of gibberella ear rot, 2.1% to 17.0% for rot kernels and 4.0% to 29.3% for incidence of F. graminearum in the kernels. Significant and positive correlations (r = 0.70) were found between incidence of F. graminearum in the kernels and rots kernels. In the third study it was watched that the kernel yield was affected by hybrids, regardless of the inoculation method. The P30F35YH and BG7049H hybrids obtained better yields, regardless of the environment and inoculation method, being classified as moderately susceptible to gibberella ear rot. The AG8025PRO, DKB250PRO2, AS1555PRO, Status, 30K64HNSR and P4285 were classified as moderately resistant considering the intensity of gibberella ear rot and the involvement of resistance to colonization of the stigma (tip of the ear) and kernel resistance. There were significant and positive relationship between productivity with injection in the silk-channel (r = 0.93) and wounding the kernels with productivity (r = 0.94) in environment 1 and 2. It was found that although there are no resistant hybrids there is genetic variability and the involvement resistance of stigma mechanisms and the kernels, and the giberella ear rot affected more kernel quality than quantity / A podridão de giberela causada pelo fungo Fusarium graminearum, provoca danos na produtividade e qualidade de grãos de milho sendo doença de difícil controle devido à indisponibilidade de híbridos resistentes e baixa eficácia de práticas culturais. Foram realizados três estudos, o objetivo do primeiro foi comparar quatro métodos de inoculação (injeção no canal do estilo-estigma, aspersão nos estigmas, ferimento dos grãos no centro da espiga e deposição na bainha da folha da espiga) e um tratamento controle em quatro híbridos (P30R50H, P30B30H, P3989, P32R48H) com base na severidade da doença e produtividade de grãos; o segundo estudo determinou à reação de 20 híbridos a podridão de giberela pela quantificação da incidência da doença, grãos ardidos e F. graminearum nos grãos e suas correlações; e o terceiro estudo caracterizou a resistência genética de híbridos à podridão de giberela pela elaboração de uma escala de reação envolvendo classes e intensidade de doença e suas correlações com produtividade de grãos e métodos de inoculação. Os experimentos do primeiro estudo foram conduzidos na safra 2011/12 em Cruz Alta, RS e Lages, SC. Os experimentos do segundo e terceiro estudos foram realizados em dois ambientes na safra agrícola 2012/13, em Cruz Alta. O delineamento dos experimentos foi em blocos casualizados com tratamentos arranjados num fatorial com quatro repetições. No primeiro estudo houve diferença na severidade da doença entre híbridos e métodos de inoculação. O híbrido P32R48H apresentou maior severidade da doença, diferindo dos demais. Inoculações no canal do estilo-estigma e ferimento dos grãos proporcionaram maior severidade da doença e menores produtividades de grãos. No segundo estudo o método de ferimento dos grãos proporcionou acréscimos superiores a 40% na incidência de espigas giberelas, incidência de grãos ardidos e incidência de F. graminearum nos grãos quando comparado ao método de injeção no canal do estilo estigma. Houve efeito de híbridos e os valores foram de 37,2% a 95,6% para incidência de espigas giberelas, 2,1% a 17,0% para grãos ardidos e 4,0% a 29,3% para incidência de F. graminearum nos grãos. Correlações significativas e positivas (r = 0,70) foram obtidas entre incidência de F. graminearum nos grãos e grãos ardidos. No terceiro estudo foi observado que a produtividade de grãos foi afetada pelos híbridos, independentemente do método de inoculação. Os híbridos P30F35YH e BG7049H obtiveram melhores produtividades, independente do ambiente e método de inoculação, sendo classificados como moderadamente suscetível à podridão de giberela. Os híbridos AG8025PRO, DKB250PRO2, AS1555PRO, Status, 30K64HNSR e P4285 foram classificados como moderadamente resistente, considerando a intensidade de giberela e o envolvimento da resistência à colonização do estigma (ponta da espiga) e resistência do grão. Houve relações significativas e positivas entre produtividade com injeção no canal do estilo-estigma (r = 0,93) e produtividade com ferimento dos grãos (r = 0,94) nos ambientes 1 e 2. Foi constatado que apesar de não haver hibridos resistentes existe variabilidade genética e o envolvimento de mecanismos de resistência do estigma e dos grãos, e que as podridões de giberela afetaram mais a qualidade dos grãos do que a quantidade

Gibberella zeae (anamorph Fusarium graminearum) is a homothallic ascomycete pathogen that is responsible for causing Fusarium head blight (FHB) of wheat and small grains. In addition to causing a reduction in yield, harvested grain is frequently contaminated with trichothecene mycotoxins that are harmful for human and animal health. Use of wheat varieties with resistance to FHB is an important strategy to lower its impact. In order to produce varieties with durable resistance, we must understand the origin and degree of genetic diversity present in the pathogen population. In my research, I focused my efforts on an investigation of the role of mating and sexual development in the generation of genotypic and phenotypic variability in G. zeae. The goal of one part of my work was to develop new genetic markers that can be used to monitor out-crossing and genetic diversity in the population. I also optimized gene deletion protocols for G. zeae so that I could produce mutant and control strains to address my research hypothesis that MAT genes play a direct role in pathogenicity. Application of novel repetitive RFLP probes to a group of G. zeae isolates originating from and near Kentucky revealed a surprisingly high degree of diversity in these local populations. Diversity between locations was greater than that within locations, suggesting the relative importance of local inoculum sources. The probes were also useful as genetic markers for segregation analysis. I crossed two genetically closely related, and commonly used, laboratory strains of G. zeae and found that this resulted in transgressive segregation for both aggressiveness and toxigenicity. I showed that the very high and very low levels of aggressiveness and toxigenicity in transgressive segregants are heritable. I also showed that selfing produced a higher degree of diversity in these traits among the progeny than was observed among conidial progeny. This suggests the presence of epigenetic factors that impact pathogenicity. Sexual behavior in G. zeae is under the control of MATing type genes. I deleted the complete MAT1 locus, and the MAT1-1-1, and MAT1-2-1 genes separately. Deletion of each of the targeted sequences produced the expected shifts in fertility phenotype. The mat1KO strains became asexual, while mat1-1-1KO and mat1-2-1KO strains shifted to obligate heterothallism. Deletion of the MAT1-1-1 and MAT1-2-1 genes had a negative effect on aggressiveness and mycotoxin production in planta, but deletion of the complete MAT1 locus had no effect. The set of mutant and ectopic control strains that I generated will be a useful asset that will be made available to the research community.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2016-08-22T11:29:51Z No. of bitstreams: 1 texto completo.pdf: 956629 bytes, checksum: 1096a842439a74fe8b122975f58e42bd (MD5) / Made available in DSpace on 2016-08-22T11:29:51Z (GMT). No. of bitstreams: 1 texto completo.pdf: 956629 bytes, checksum: 1096a842439a74fe8b122975f58e42bd (MD5) Previous issue date: 2016-02-23 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Os fungicidas são normalmente aplicados para o manejo da giberela com o objetivo de garantir a produtividade e reduzir níveis de micotoxinas associadas à doença. Vários ingredientes ativos estão disponíveis no Brasil, sendo que os mais comumente recomendados pertencem ao grupo dos inibidores da demetilação (DMI) e benzimidazóis (MBC), ou usados em mistura com fungicidas do grupo das estrobilurinas (QoI). Dada a importância crescente de epidemias de giberela no Brasil, o uso de fungicidas vêm aumentando consideravelmente e, geralmente, duas aplicações têm sido recomendadas. Parcelas de campo em 16 municípios na região de Guarapuava, Sul do Paraná foram monitoradas durante quatro anos (2011 a 2014) e um total de 227 isolados semelhantes a espécies do complexo de espécies de Fusarium graminearum foram obtidos a partir de espigas de trigo sintomáticas. Uma amostra de 173 isolados foi identificada quanto ao genótipo tricoteceno com base em PCR. Entre eles, 67% foram identificados como 15- (A)cetil-desoxinivalenol(DON), 29% do nivalenol (NIV) e 4% como genótipo 3-ADON. Uma amostra de 35 isolados foi selecionada para determinar a concentração dos fungicidas tebuconazol e carbendazim que reduz em 50% do crescimento de micelial (EC 50 ), usando as seguintes doses (0; 0,5; 1,0; 2,0 e 4,0 μg / ml). As medias estimadas de EC 50 , independente do genótipo, foram 0,39 μg/ml (0,0004 a 3,0) e 1,25 μg/ml (0,91 a 2,65) para tebuconazol e carbendazim, respectivamente. As medias de EC 50 foram 1,32 μg/ml e 1,21 μg/ml para carbendazim e 0,58 μg/ml e 0,05 μg/ml para tebuconazol, para os isolados 15-ADON e NIV ou 3-ADON, respectivamente. O teste não paramétrico Kolmogorov-Smirnov mostrou diferença entre os isolados 15-ADON e os NIV+3-ADON quanto ao tebuconazol (P = 0,002), mas não para o carbendazim (P = 0,514). As doses discriminatórias de 1,4 μg/ml e 2,0 μg/ml foram usadas para identificar isolados menos sensíveis à carbendazim e tebuconazol, respectivamente, em toda coleção. Baseado nessas doses, dois isolados menos sensíveis ao tebuconazol e seis isolados menos sensíveis ao carbendazim foram encontrados, sendo todos originários do munícipio de Guarapuava ao longo de três safras e possuem o genótipo 15-ADON. Houve correlação significativa entre os valores de EC 50 dos dois fungicidas (r = 0,45; P = 0,007). Um único isolado apresentou os maiores valores de EC 50 para ambos fungicidas e apresentou uma mutação (R31K) no gene cyp51A. A presença da reduzida sensibilidade e um isolado resistente, reportado pela primeira vez no Brasil, sugerem que a seleção pode conduzir mudanças nas populações com consequências no manejo da doença. Estudos futuros devem se concentrar em atributos de fitness, competitividade e experimentos in vivo para verificar se a eficácia do fungicida é prejudicada na presença de isolados menos sensíveis. Em estudos de controle químico da giberela, resultados sobre a eficácia têm sido inconsistentes e variáveis para um mesmo fungicida. Uma revisão sistemática de estudos de eficácia dos fungicidas pode ser útil para apontar para os produtos mais eficazes e cenários onde a eficácia pode ser melhorada. Os objetivos deste trabalho foram a) revisar sistematicamente estudos de controle químico para o manejo da giberela realizados no Brasil durante os últimos 15 anos (2000-2015), publicados em revistas com e sem corpo editorial e b) conduzir uma metanálise da eficácia de controle e retorno em produtividade de fungicidas selecionados. Uma busca na literatura identificou 18 estudos que relataram a severidade da giberela (o mesmo que índice giberela) para pelo menos um tratamento com fungicida em comparação com um tratamento controle sem o fungicida, e também uma medida da variância amostral do ensaio. Apenas três e dois estudos relataram valores médios de grãos giberelados e deoxinivalenol (DON), respectivamente. Os estudos foram analisados e oito tratamentos com fungicidas, com pelo menos dez entradas foram selecionados para o estudo de metanálise. Foram incluídos os tratamentos com os fungicidas dos grupos: DMI (tebuconazol, metconazol e propiconazol) e MBC (carbendazim) formulados individualmente e uma pré-mistura de DMI + QoI (azoxistrobina + tebuconazol, piraclostrobina + metconazol, trifloxistrobina + prothioconazol e trifloxistrobina + tebuconazol). Foram testadas uma aplicação (×1) e duas aplicações (×2), sendo a primeira na floração plena e a segunda dez dias após. Um modelo metanalítico multivariado foi ajustado para o log das médias para cada fungicida e o tratamento controle de cada ensaio. A diferença nas estimativas médias do log da severidade entre um tratamento com fungicida e o controle foram usadas para calcular a razão de resposta média (R) e então transformadas para porcentagem de controle (C) e porcentagem de retorno em produtividade (P). A média de C e P variou de 48,94 a 70,39% e de 7,41 a 21,29%, respectivamente. Metconazol (×2) teve o melhor desempenho para reduzir a severidade da doença (C = 70,39%) e tebuconazol (×1) em retorno em produtividade (P = 21,29%). Propiconazol foi o menos eficaz entre os DMIs (C = 48,94%), seguido de carbendazim (C = 53,90%). Em geral, os fungicidas mais eficazes resultaram em maior retorno de rendimento. Quando os grupos de fungicidas foram comparados, em geral, os DMIs e as misturas tiveram o melhor desempenho, especialmente em ensaios conduzidos sob elevada pressão da doença (> 7% no tratamento controle). Os resultados mostraram que a eficácia de controle média em estudos brasileiros quando se utiliza duas aplicações são mais elevadas do que quando se utiliza apenas uma aplicação, como relatado em outro estudo de metanálise na América do Norte. Estudos futuros deverão centrar-se no efeito sobre os níveis de DON, especialmente para as misturas, que estão estrategicamente utilizadas para o controle de doenças foliares. / Fungicides are commonly applied for managing Fusarium head blight (FHB) with the aim of protecting yield and improving grain quality, but especially targeting mycotoxin reduction. Several active ingredients are available in Brazil and the most common belong to the demethylation-inhibitor (DMI) and benzimidazoles (MBC) groups, with the former commonly used in mixture with fungicides of the quinone-outside inhibitor (QoI) group. Given the increasing importance of FHB epidemics in Brazil, the use of fungicides is increasing considerably and usually two sprays are recommended. Experimental trials where fungicides have been applied for FHB control were monitored in 16 municipalities at the Guarapuava region, Southern Paraná, during four years (2011 to 2014). A total of 227 isolates resembling those of the Fusarium graminearum species complex were obtained from wheat heads with FHB symptoms. Of those, 173 isolates were assigned to trichothecene genotypes based on PCR assays. A random sample of 35 isolates from 2011 to 2013 was selected to determine the concentration of tebuconazole and carbendazim fungicides that reduces 50% of the mycelial growth (EC 50 ). The following doses were used: 0, 0.5, 1.0, 2.0 and 4.0 μg/ml. In the PCR assays, most isolates (67%) were of the 15-(A)cetyl-deoxynivalenol (DON), 29% of the nivalenol (NIV) and 4% of the 3-ADON genotype. The estimated means of the EC 50 , irrespectively of the genotype, were 0.39 μg/ml (0.0004 to 3.0) and 1.25 μg/ml (0.91 to 2.65) for tebuconazole and carbendazim, respectively. The mean EC 50 were 1.32 μg/ml and 1.21 μg/ml for carbendazim and 0.58 μg/ml and 0.05 μg/ml for tebuconazole, for isolates of the 15-ADON and NIV + 3-ADON isolates, respectively. A Kolmogorov-Smirnov non-parametric test suggested that the sensitivity to tebuconazole of the 15-ADON and NIV+3-ADON isolates were sampled from populations with different distributions (P = 0.002), but not for carbendazim (P = 0,514). Discriminatory doses of the 1.4 μg/ml and 2.0 μg/ml were used to screen for less- sensitive isolates in the whole sample. Based on those doses, two tebuconazole-less sensitive and six carbendazim-less sensitive isolates were found, all were originated from Guarapuava municipality across three growing seasons and possessed the 15-ADON genotype. There was as significant correlation between the EC 50 values of tebuconazole and carbendazim (r = 0.45; P = 0,007). One isolate exhibited the highest EC 50 values for both fungicides and showed a mutation (R31K) in the cyp51A gene. The presence of less sensitive and a resistant isolate reported for the first time in Brazil suggest that selection may drive changes in the populations with impact in disease management. Future studies should focus on the attributes of fitness, competitiveness and in vivo assays to check whether fungicide efficacy is hindered in the presence of resistant isolates. Furthermore, results on fungicide efficacy have been inconsistent and variable for a same fungicide. Quantitative summary of fungicide data available in the country is lacking and could be useful to point to the most efficacious products and scenarios where efficacy can be improved. The objectives of this work were to a) systematically review the peer- and non- peer reviewed literature on chemical control studies for FHB management conducted in Brazil during the last 15 years and b) conduct a meta-analysis of the control efficacy and yield return of selected fungicides. A search in the literature identified 18 studies that reported FHB severity (same as FHB index) for least one fungicide treatment compared to a non-treated check, and also a measure of sampling variance in the trial. Only three and two studies reported mean values of Fusarium-damaged kernels (FDK) and deoxyninvalenol (DON), respectively. Studies were scrutinized and eight fungicide treatments with at least 10 entries were selected for the meta-analysis study. They included DMI (tebuconazole, metconazole and propiconazole) and MBC (carbendazim) fungicide treatments formulated solo and a pre-mix of DMI+QoI (azoxystrobin + tebuconazole, pyraclostrobin + metconazole, trifloxystrobin + prothioconazole and trifloxystrobin + tebuconazole). One (×1) and two applications (×2) were tested, the first at full flowering and the second 10 days later. A multivariate meta-analytic model was fitted to the log of the means for each fungicide and check treatment of the trial. The difference in the mean estimates of the log of severity between a fungicide treatment and the check were used to calculate the mean response ratio represented by percentage control efficacy (C) and percentage yield return (P). Mean C and P ranged from 48.94 to 70.39% and from 7.41 to 21.29%, respectively. Metconazole (×2) performed best to reduce disease severity (C = 70.39%) and tebuconazole (×1) to increase yield (P = 21.29%). Propiconazole was the least efficacious among the DMIs (48.94%), followed by carbendazim (53.9%). In general, the most efficacious fungicides resulted in higher yield return. When fungicide groups were compared, the DMIs and the mixtures performed best in general, especially in trials conducted under high disease pressure (>7% in the check treatment). Results showed that mean control efficacy in Brazilian studies when using two sprays are higher one spray, as reported in other meta-analysis study in North America. Future studies should focus on the effect on DON levels, especially for the mixtures, which are strategically used for the control foliar diseases.

Doctor of Philosophy / Department of Plant Pathology / Robert L. Bowden / John F. Leslie / Gibberella zeae is an ascomycete filamentous fungus and the major cause of Fusarium head blight, also called scab, in small grains. This dissertation contains three related studies of G. zeae. In the first, the genetic map was aligned with the first assembly of the genomic sequence released by The Broad Institute (Cambridge, MA). Approximately 99% of the sequence was anchored to the genetic map, indicating the high quality of the sequence assembly and validity of the genetic map. The alignments grouped the linkage groups and supercontigs into four sets, which is consistent with the hypothesis that there are four chromosomes in this fungus. In the second, the sex pheromone precursor genes (ppg1 and ppg2) and the pheromone receptor genes (pre1 and pre2) were identified and characterized. Deletion of ppg1 or pre2 ([Delta]ppg1 or [Delta]pre2 strains) reduced the number of perithecia produced by self-fertilization, but did not completely block perithecial formation. The proportion of crosses resulting from outcrossing increased when the [Delta]ppg1 strains were used as the female in crosses with male strains containing an intact ppg1 gene. [Delta]ppg2 and [Delta]pre1 mutants had no discernable effect on morphological phenotype or self-fertilization. Thus, one of the pheromone/receptor pairs (ppg1/pre2) found in many Ascomycetes has a role in, but is not essential for, selfing or outcrossing in G. zeae, whereas the other pheromone/receptor pair (ppg2/pre1) no longer has a detectable function in sexual reproduction. In the third study, spore germination of G. zeae was tested in the presence of α- factor-like pheromone peptides of G. zeae or N. crassa. The pheromone peptide of N. crassa more efficiently inhibited spore germination than did the peptide from G. zeae. Arginine and lysine residues were the most important determinants in blocking spore germination. In conclusion, this research has validated the genetic map and the genomic assembly of G. zeae, characterized sex pheromone functions and characterized pheromone peptide ability to inhibit spore germination. The pheromone peptides of G. zeae and N. crassa may be useful as control agents for G. zeae and pheromone peptide efficacy might be further enhanced by judicious substitutions for some of the amino acids.

Fusarium head blight (FHB) is one of the most economically important diseases of small grains and continues to impact crops when environmental conditions are favorable to Gibberella zeae (Fusarium graminearum), the causal agent of the disease. Corn residues are considered to be primary sources of inoculum for epidemics of FHB. Therefore, knowledge of the movement of Gibberella zeae from a local source of infested corn residue is critical to the management of FHB in wheat and barley. Previous research made significant progress in defining the spatial dissemination of inoculum sources of G. zeae within agricultural fields, but was unable to clearly distinguish between within-field and background sources. Using amplified fragment length polymorphism, released clones of G. zeae were tracked within wheat and barley fields. This strategy allowed the distinction between the contributions of released clones to FHB, compared to that of background inocula. Corn residue infested with clones of G. zeae was placed into small replicated plots in winter wheat fields in New York and Virginia in 2007 and 2008 and wheat spikes were collected at 0, 3, 6, and ≥24 m from the inoculum sources. Recovery of released clones decreased an average of 90% between 3 and 6 m from inoculum sources. Various amounts of corn residue infested with a single clone of G. zeae were placed into small replicated plots in winter wheat and barley fields in Virginia from 2008 to 2010. The use of minimal or conventional tillage and a moderately resistant cultivar of wheat or barley may reduce the contribution of within-field inocula to FHB; however, environmental conditions play an important role in the effectiveness of these management strategies. With the increase of corn production due to incentives for ethanol-based fuel, overwintering sites for G. zeae on corn residue are likely to increase. Our work contributes to an increased understanding of the influence of overwintered corn residue to FHB which will also direct future research on how to reduce the inoculum potential from within-field sources. / Ph. D.

Submitted by Gabriela Lopes (gmachadolopesufpel@gmail.com) on 2016-10-03T17:35:05Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Taiane_Viana.pdf: 904492 bytes, checksum: 30037558592bcb3fdf60ab01d643c574 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2016-10-06T20:46:17Z (GMT) No. of bitstreams: 2 Dissertacao_Taiane_Viana.pdf: 904492 bytes, checksum: 30037558592bcb3fdf60ab01d643c574 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-10-06T20:46:17Z (GMT). No. of bitstreams: 2 Dissertacao_Taiane_Viana.pdf: 904492 bytes, checksum: 30037558592bcb3fdf60ab01d643c574 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2013-12-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / O cultivo do trigo no Brasil é marcado pelo elevado número de doenças capazes de causar danos à produção. Entre as principais doenças que afetam o trigo está a giberela ou fusariose da espiga, causada pelo agente etiológico Gibberella zeae (Schw.) Petch, (anamorfo Fusarium graminearum Schwabe) que causa impacto negativos não somente à produtividade mas também a qualidade dos grãos produzidos. As condições ambientais para que a infecção ocorra carac terizam-se com temperaturas de 20-25°C e duração do molhamento foliar de, no mínimo, 48 horas consecutivas. O controle químico é realizado via aplicação de fungicidas a partir do inicio da floração estendendo-se até o final do florescimento, sendo que este tipo de controle caracteriza-se pela sua menor eficiência quando comparado com o controle de doenças foliares. Caso as condições climáticas impeçam a realização das aplicações de fungicidas no período indicado, o controle fica muito dificultado, por outro lado, se não ocorrer clima favorável à infecção, não se justifica o tratamento. A ineficiência dos tratos culturais no controle de giberela, associado à natureza quantitativa da característica de resistência, e o fato de sofrer alta influência do ambiente resultam na necessidade de identificar fontes efetivas de resistência que possam ser utilizadas em programas de melhoramento de trigo. De fato, existem culturas onde o controle das doenças mais importantes ocorre, quase que exclusivamente, por meio da resistência sendo que a identificação de genotipos que contenham resistência é um dos primeiros passos para selicioná-los. Desta forma, é fundamental o conhecimento de cultivares que apresentem níveis de resistência a giberela e métodos que possam identificar estas cultivares. Frente ao exposto o objetivo deste trabalho foi: i) separar genótipos suscetíveis e que contenham diferentes níveis de resistência utilizando avaliações morfológicas e moleculares; ii) verificar a resistência à giberela nas cultivares Frontana e Pampeano. Para isso foram utilizadas avaliações de características morfológicas citadas na literatura e associadas a resistência à giberela, assim como, o meprego de primers descritos na literatura significativamente associados à QTL de resistência à giberela. Como resultado do estudo espera-se auxiliar no desenvolvimento de ferramentas que possam ser utilizadas em programas de melhoramento de trigo visando facilitar a identificação de fontes de resistência. / The cultivation of wheat in Brazil is marked by the large number of diseases that can damage the production. Among the major diseases affecting wheat is wheat scab or Fusarium head blight (FHB), caused by the agent Gibberella zeae (Schw.) Petch (anamorph Fusarium graminearum Schwabe ) that causes negative impact not only on productivity but also the quality of the grain produced. The environmental conditions for infection are temperatures between 20-25°C and 48 consecutive hours of leaf wetness. Chemical control is performed via application of fungicides from the start extending until the end of flowering, and this type of control is characterized by its low efficiency when compared to the disease control. If the weather conditions prevent the aplication of fungicide applications within this period, there is no possibility control, on the other hand, if it does not favorable environment for infec tion treatment is not justified. The inefficiency of cultural control of FHB, associated with the quantitative trait (QTL- Quantitative traits loci)of resistance, and the fact of suffering high influence of the environment results in the need to identify effective sources of resistance that can be used in wheat breeding programs. In fact, there are cultures where control of major diseases occurs almost exclusively through resistance where the identification of genotypes containing resist ance is one of the first steps in their selection. Thus it is crucial to understand cultivar present levels of resistance to FHB and methods that can identify these cultivars.The objective of this study was: i) separate susceptible and containing different levels of resistance using molecular and morphological evaluations; ii) determine FHB resistance using cultivars Frontana and Pampeano as a source of resistance. Using morphological assessments cited in the literature and associated with resistance to FHB, as, primers as described in the literature as significantly associated with QTL for resistance to FHB. As a result of the study one expects to develop tools that can be used to facilitate the identification of sources of resistance.

Master of Science / Department of Plant Pathology / Robert L. Bowden / John F. Leslie / Fusarium graminearum Schwabe sensu lato (teleomorph: Gibberella zeae (Schwein.) Petch), a homothallic ascomycete fungus, is the causal agent of Fusarium head blight (FHB) of wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), and other small grains. FHB occurs worldwide and serious outbreaks have been reported in North America, South America, Asia, and Europe. According to the phylogenetic species concept (PSC), F. graminearum is composed of at least 15 phylogenetic lineages known as the Fusarium graminearum species complex. Although F. graminearum is homothallic, some members of different phylogenetic lineages are known to intercross in the laboratory. It has been suggested that F. graminearum sensu lato fits the biological species concept (BSC). According to the BSC, “species are groups of actually or potentially interbreeding natural populations, which are reproductively isolated from other such groups”. Previous reports of intercrossing were qualitative, so the degree of reproductive isolation, if any, is not clear. Since intrinsic reproductive isolation is the key criterion to identify species by the BSC, more detailed quantitative information is needed. Chromosome rearrangements between fungal strains may reduce fertility in sexual crosses through the production of genetically inviable recombinant progeny. As such, rearrangements can be important postzygotic reproductive barriers between species. Following methods used in Neurospora crassa, ascospore tetrads were analyzed for patterns of ascospore viability. Crosses were made with three lineage 7 (F. graminearum sensu stricto according to PSC) strains as female. Each female was a MAT1-2 knockout mutant that rendered it obligately heterothallic. Males were several members of lineages 6 (F. asiaticum according to PSC) and lineage 7. Crosses with lineage 7 males formed complete asci with 8 ascospores indicating that their genomes are isosequential with the testers. Crosses with one strain from lineage 6 with two known inversions produced asci containing 8, 6, and 4 ascospores, consistent with it not being isosequential. However, three other strains of lineage 6 appeared to be isosequential with the testers. Therefore, chromosome rearrangements did not appear to be common to strains of lineage 6 and probably do not contribute significantly to reproductive isolation of lineage 6 and lineage 7. Interlineage fertility studies with the three lineage 7 tester strains were performed to quantify interlineage fertility parameters including the total number of ascospores produced, perithecial density, and perithecium internal development scores. All lineage 7 female testers successfully crossed to all 23 male strains from lineages 1 to 9. For total ascospore production, one female tester crossed equally well with all lineages and the other two testers showed statistically significant differences for a few lineages. For perithecial density, there was a significantly lower density with all three testers when crossed with lineage 6, but the other lineages were not statistically different from lineage 7. For perithecial development, there was large variation for every lineage. Therefore, in the crosses with reduced fertility, the reduction can be attributed to a postzygotic effect since mature perithecia and asci developed. All of the tested lineages of the Fusarium graminearum species complex can produce viable progeny with F. graminearum lineage 7, which was the taxonomic type of the original species before it was split into phylogenetic species. There are a few examples of reduced fertility with two lineage 7 testers, the remaining tester crossed equally well with all lineages. Therefore members of lineages 1-9 all should be considered members of Fusarium graminearum according to the BSC. The existing female testers could be used to identify members of the F. graminearum clade by performing test crosses in the laboratory. The PSC and BSC species concepts do not agree for this group of fungi. This disagreement indicates that the F. graminearum species complex is in the early stages of speciation. The lack of intrinsic reproductive barriers supports the hypothesis that these lineages have developed in geographic isolation. As the lineages have apparently been brought together through global trade, interlineage hybrids have been reported in the field. The discrepancy between PSC and BSC will eventually be resolved by whether the lineages fuse or remain separate in nature. Even if the lineages remain separate, this study demonstrates the potential for gene flow between lineage 7 and lineages 1 through 9.

Gibberella zeae (anamorph: Fusarium graminearum) is the causal agent of fusarium head blight (FHB) and maize ear rot, two major diseases of wheat and corn in Eastern Canada. / In Quebec, Microsphaeropsis sp., an antagonist of Venturia inaequalis, the causal agent of apple scab, was isolated from the apple leaf litter. This fungus, well adapted to Quebec climate, can reduce the initial inoculum of V. inaequalis. FHB and Gibberella ear rot are similar to apple scab because the major inoculum source comes from melanized structures produced on crop residues. Consequently, we evaluated the potential of Microsphaeropsis sp. (isolate P130A) to inhibit ascospore production of G. zeae when applied to crop residues as post harvest or pre-planting applications. Under in vitro conditions, the antagonist significantly reduced ascospore production on wheat and corn residues, when applied prior to (82% and 92% respectively) or at the same time as the pathogen (36% and 58% respectively). Under field conditions, the antagonist had no effect on the pattern of perithecia maturation but significantly reduced the number of ascospores produced on two sampling dates, May 1998 and July 1999. (Abstract shortened by UMI.)