Practical resistance of Erysiphe necator to quinone outside inhibitors (QoIs) is now widespread, and resistance to demethylation inhibitors (DMIs) has also developed.  The goal of this research was to characterize fungicide resistance by elucidating resistance mechanisms and determining its stability.  QoI resistance persisted for several years in a field population after QoI application ended.  Resistant isolates were highly competitive in mixed populations in competition assays under laboratory conditions, indicating a lack of fitness cost.  In one competition trial under field conditions, resistance frequency declined, possibly due to spore migration and influx of background inoculum, but in a second trial, it did not decline.  Double resistance to QoI and DMI was detected and DMI application may have been partially responsible for maintaining QoI resistance in the field.  One isolate with QoI resistance but an undetectable level of the major QoI mutation was shown to be heteroplasmic -- resistant strains could be selected from this isolate. DMI resistance mechanisms in E. necator included the Y136F mutation in CYP51 and cyp51 over-expression.  The first mechanism was present in almost all isolates with substantial levels of resistance, and cyp51 expression level was correlated with resistance level.  Three cyp51 genotypes were detected.  Wildtype isolates with the TAT genotype were sensitive to DMIs, while isolates with increased resistance had either a TTT or TWT genotype; TWT indicated the presence of both wildtype and mutant alleles.  Cyp51 was expressed 1.4 to 19 times more in mutants than in wildtype.  It is not known whether the significant differences in cyp51 expression level among isolates and among genotype groups are due to gene copy number variation.  DMI resistance was found to decline after years of subculturing, and the decline appeared to occur after a few culture transfers of field samples on fungicide-free host leaves.  The observed decline, together with the finding that isolates could be "trained" to increase resistance, and may be slightly induced in cyp51 expression when successively challenged to grow in increasing fungicide concentration, indicate instability of DMI resistance. / Ph. D.

The honey bee is a widely managed crop pollinator that provides the agricultural industry with the sustainability and economic viability needed to satisfy the food and fiber needs of our society. Excessive use of agrochemicals such as the acaricides coumaphos and tau-fluvalinate, and the fungicide, chlorothalonil is implicated in the reduced number of managed bee colonies available for crop pollination services. Here, I report the effects of pesticide exposures on the nutritional and immune health of the honey bee. Total protein concentration was significantly reduced in the coumaphos- and chlorothalonil-treated individuals compared to the pesticide-untreated bees. Total carbohydrate concentration was significantly reduced in the tau-fluvalinate-, coumaphos-, and chlorothalonil-treated individuals compared to the pesticide-untreated bees. Total lipid concentration was significantly decreased in the chlorothalonil-treated individuals compared to the pesticide-untreated bees. Body weight was significantly reduced for the tau-fluvalinate-, coumaphos-, and chlorothalonil-treated individuals, respectively, compared to the pesticide-untreated bees. Head width was significantly reduced for the chlorothalonil-treated individuals whereas the wing length was significantly reduced for the coumaphos and chlorothalonil-treated individuals, respectively, compared to the pesticide-untreated bees. Phenoloxidase activity was significantly increased in the coumaphos-treated individuals compared to the pesticide-untreated bees. Glucose oxidase activity was significantly increased in the chlorothalonil-treated individuals compared to the pesticide-untreated bees. While more research is needed to verify the observed effects of the pesticides on the nutritional and immunity health of the honey bee, it is important for beekeepers to consider alternative methods for control of varroa mites and the use of fungicides near their colonies. / Master of Science in Life Sciences

The restoration of large-scale heterogeneous landscapes is typically approached using seed-based restoration as a cost-effective means to achieve project goals and objectives. However, seeding is often met with low survival rates primarily driven by low seedling emergence and establishment. This dissertation investigates opportunities to enhance restoration focusing on 1) seed treatments, 2) plant materials, and 3) a synthesis of seeding strategies. • Fungal pathogenesis can cause high seedling mortality. Fungicide seed coatings used in the agriculture industry are novel to wildland restoration. We assess the effect a fungicide seed coating has on the recruitment of Pseudoroegneria spicata seedlings across a broad range of sagebrush-steppe sites. • Sourcing appropriate plant material for restoration seeding is fundamental to long-term restoration success. The Association of Seed Certification Agencies offers a series of certification schemes to facilitate the rapid production, procurement, and distribution of native seeds for restoration. Results from a multi-year study evaluate the recruitment success of three P. spicata germplasms. • The complexities of seed-based restoration are reviewed, exploring the interplay between native plant materials, seed enhancement technologies, and restoration outcomes. This synthesis provides a comprehensive overview of the current state of knowledge, research gaps, and avenues for future restoration research.

A videira é uma das plantas mais antigas cultivadas pela humanidade, sendo que no Brasil a uva é a terceira fruta com maior volume de produção, atrás apenas do cultivo das bananas e das laranjas. Apesar da produção rentável, principalmente aos pequenos produtores, o parreiral é susceptível a várias doenças cujo manejo compromete até 59% dos gastos do produtor. No estado de São Paulo, dentre as doenças, três têm destaque: a antracnose (causada pelo Sphaceloma ampelinum), o míldio da videira (causado pelo Plasmopara viticola) e a ferrugem (causada pelo Phakopsora euvitis). Os produtores utilizam controle químico de forma intensa e preventiva, chegando a 100 aplicações de fungicidas em um ciclo de até 120 dias. Os principais fungicidas utilizados são os inibidores da quinona oxidase (QoI), que agem impedindo o transporte de elétrons do citocromo b ao citocromo c1 na cadeia respiratória da mitocôndria. Porém, existem relatos de resistência ao fungicida aplicado no campo em diversos países. As substituições G143A, G137R e F129L na sequência da proteína citocromo b impedem que o fungicida se ligue ao seu sítio alvo. As mutações que levam às substituições estão localizadas em uma das regiões chamada hotspot do gene citocromo b (cytB). Visto que, pela carência de estudos, a resistência genética a esses fungicidas nunca foi relatada no Brasil, o objetivo principal desse trabalho foi sequenciar e caracterizar a região hotspot em isolados de míldio, ferrugem e antracnose provenientes de parreirais do estado de São Paulo. Foram selecionados 35 isolados de 11 locais diferentes; desses, 11 isolados de míldio foram considerados geneticamente resistentes, pois apresentam a mutação para o resíduo alanina na posição 143, e 4 isolados foram considerados geneticamente sensíveis. Os dois isolados de ferrugem selecionados também foram considerados geneticamente sensíveis. Pela estratégia de Genome Walking foi possível sequenciar 65% do gene cytB de um dos isolados brasileiros de P. viticola; foram encontrados poucos polimorfismos e nenhum íntron na sequência analisada. Os resultados obtidos com esse estudo podem servir de suporte para a tomada de decisões de manejo mais adequadas para a realidade da viticultura brasileira; além disso, são importantes para futuros estudos sobre a evolução do patógeno com a pressão seletiva exercida pelos fungicidas. / Grapevine is one of the most ancient cultivated plants and its fruit, grape, is notably important in Brazil, since it is the third most produced, only behind banana and citrus. Although it is rentable especially to smallholders, the vineyard is often attacked by several pathogens and the damages induced by them can compromise up to 59% of the producers\' expenses in order to keep the diseases under control. In Sao Paulo state there are three important diseases that attack vineyards: anthracnose (caused by Sphaceloma ampelinum), downy mildew (caused by Plasmopara viticola) and rust (caused by Phakopsora euvitis). Pest management practices used by the producers relies on intensive and preventive use of fungicides, in which the culture is sprayed 100 times per vineyard\'s growth cycle (that last approximately 120 days). One of the most used fungicides are the quinone oxidase inhibitors (QoI), that act by blocking the electron transport chain at the mitochondria binding at the Qo site of the cytochrome b (cytB) complex. However, there are several reports of the presence of resistant strains in different countries. Resistance is caused by the aminoacids substitutions F129L, G137R and G143A in the cytochrome b protein sequence, that prevent the fungicide molecule binding to its target site. The mutations in the cytB gene that lead to these substitutions are harbored in a region called hotspot for fungicide resistance. Since this type of study was never reported in Brazil, the main purpose of this work was to sequence and characterize the hotspot region of different isolates from anthracnose, downy mildew and rust. Thirty five isolates from eleven different locations were choosen for the study. Eleven of them harbored the mutation that lead to the substitution G143A; these were then considered genetically resistant to the QoI fungicides. On the contrary, four downy mildew and the two rust isolates were considered sensitive to the QoI fungicides, since none of the aminoacids substitutions were observed. Also, by using a technique named Genome Walking it was possible to sequence 65% of cytB gene from a Brazilian downy mildew isolate. In this sequence were found few polymorphisms and none intron. These study findings are unique for Brazilian isolates and might be useful to provide reliable support for the pest management decisions regarding the reality that is found at the vineyards in Brazil. Furthermore, the results presented here are important to the comprehension of pathogen\'s evolution when suffering from a selective pressure caused by the intensive use of fungicides.

A videira é uma das plantas mais antigas cultivadas pela humanidade, sendo que no Brasil a uva é a terceira fruta com maior volume de produção, atrás apenas do cultivo das bananas e das laranjas. Apesar da produção rentável, principalmente aos pequenos produtores, o parreiral é susceptível a várias doenças cujo manejo compromete até 59% dos gastos do produtor. No estado de São Paulo, dentre as doenças, três têm destaque: a antracnose (causada pelo Sphaceloma ampelinum), o míldio da videira (causado pelo Plasmopara viticola) e a ferrugem (causada pelo Phakopsora euvitis). Os produtores utilizam controle químico de forma intensa e preventiva, chegando a 100 aplicações de fungicidas em um ciclo de até 120 dias. Os principais fungicidas utilizados são os inibidores da quinona oxidase (QoI), que agem impedindo o transporte de elétrons do citocromo b ao citocromo c1 na cadeia respiratória da mitocôndria. Porém, existem relatos de resistência ao fungicida aplicado no campo em diversos países. As substituições G143A, G137R e F129L na sequência da proteína citocromo b impedem que o fungicida se ligue ao seu sítio alvo. As mutações que levam às substituições estão localizadas em uma das regiões chamada hotspot do gene citocromo b (cytB). Visto que, pela carência de estudos, a resistência genética a esses fungicidas nunca foi relatada no Brasil, o objetivo principal desse trabalho foi sequenciar e caracterizar a região hotspot em isolados de míldio, ferrugem e antracnose provenientes de parreirais do estado de São Paulo. Foram selecionados 35 isolados de 11 locais diferentes; desses, 11 isolados de míldio foram considerados geneticamente resistentes, pois apresentam a mutação para o resíduo alanina na posição 143, e 4 isolados foram considerados geneticamente sensíveis. Os dois isolados de ferrugem selecionados também foram considerados geneticamente sensíveis. Pela estratégia de Genome Walking foi possível sequenciar 65% do gene cytB de um dos isolados brasileiros de P. viticola; foram encontrados poucos polimorfismos e nenhum íntron na sequência analisada. Os resultados obtidos com esse estudo podem servir de suporte para a tomada de decisões de manejo mais adequadas para a realidade da viticultura brasileira; além disso, são importantes para futuros estudos sobre a evolução do patógeno com a pressão seletiva exercida pelos fungicidas. / Grapevine is one of the most ancient cultivated plants and its fruit, grape, is notably important in Brazil, since it is the third most produced, only behind banana and citrus. Although it is rentable especially to smallholders, the vineyard is often attacked by several pathogens and the damages induced by them can compromise up to 59% of the producers\' expenses in order to keep the diseases under control. In Sao Paulo state there are three important diseases that attack vineyards: anthracnose (caused by Sphaceloma ampelinum), downy mildew (caused by Plasmopara viticola) and rust (caused by Phakopsora euvitis). Pest management practices used by the producers relies on intensive and preventive use of fungicides, in which the culture is sprayed 100 times per vineyard\'s growth cycle (that last approximately 120 days). One of the most used fungicides are the quinone oxidase inhibitors (QoI), that act by blocking the electron transport chain at the mitochondria binding at the Qo site of the cytochrome b (cytB) complex. However, there are several reports of the presence of resistant strains in different countries. Resistance is caused by the aminoacids substitutions F129L, G137R and G143A in the cytochrome b protein sequence, that prevent the fungicide molecule binding to its target site. The mutations in the cytB gene that lead to these substitutions are harbored in a region called hotspot for fungicide resistance. Since this type of study was never reported in Brazil, the main purpose of this work was to sequence and characterize the hotspot region of different isolates from anthracnose, downy mildew and rust. Thirty five isolates from eleven different locations were choosen for the study. Eleven of them harbored the mutation that lead to the substitution G143A; these were then considered genetically resistant to the QoI fungicides. On the contrary, four downy mildew and the two rust isolates were considered sensitive to the QoI fungicides, since none of the aminoacids substitutions were observed. Also, by using a technique named Genome Walking it was possible to sequence 65% of cytB gene from a Brazilian downy mildew isolate. In this sequence were found few polymorphisms and none intron. These study findings are unique for Brazilian isolates and might be useful to provide reliable support for the pest management decisions regarding the reality that is found at the vineyards in Brazil. Furthermore, the results presented here are important to the comprehension of pathogen\'s evolution when suffering from a selective pressure caused by the intensive use of fungicides.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Aiming to evaluate the response of ten soybean cultivars subjected to three arrangements of plants and eight fungicide programs in terms of expression and the development of Asian rust, penetration of light on the canopy, productivity and yield components of soybean experiment was conducted during the 2008/2009 season, in Itaara / RS. In the field, cultivars FCep 55, BMX Apollo, BMX Titan, BRS 243, A 8000, CD 214, NK 70, BRS Pampa, FCep 59 and Monasca were subjected to densities off 160, 300 and 440 thousand plants ha-1 and eight programs fungicide management: P1- treatment control, P2 - epoxiconazole + pyraclostrobin with application to closing the row space and 21 days after the first application; P3 - epoxiconazole + pyraclostrobin application to closing the row space and 28 days; P4 - epoxiconazole + pyraclostrobin with application in R1 and 21 days; P5 - epoxiconazole + pyraclostrobin with applications closing the row space, 21 days after the first application and the third application 14 days after the second; P6 - tebuconazole + carbendazin with application in closing the row space and 21 days after the first application; P7 - tebuconazole + carbendazin application to close row space and 28 days after and P8 - tebuconazole + carbendazin with application in R1 and 21 days. The variables were: percentage of light penetration within the canopy, Area under the progress disease curve (AUPDC), grain yield and yield components. The experimental design was a randomized block, with 4 replications and three-way design (10 cultivars x 3 planting densities x 8 fungicide management programs). For percentage of light penetration in the canopy, BMX Apollo cultivar performed better with values close to 1% when subjected to higher density. For the variables AUPDC and productivity cultivars might be grouped as responsive, not responsive and unresponsive to reduce the plant population. The responsive cultivars for AUPDC were FCep55, BMX Apollo, BRS 243, CD 214, has had this behavior to yield the cultivars FCep 55, A 8000, BMX Titan, BRS 243 and NK 70. Management programs had the same trend for these two variables where the program with three applications of epoxiconazole + pyraclostrobin showed lower AUPDC value and higher yield. The number of branches per plant showed that all cultivars reduce the production of branches when exposed to high density, which resulted in condensing the grain on the main stem, especially the cultivar BMX Apollo that concentrated 99.84% of the grain on the main stem and the higher density was only 0.02 branches for plant. For the yield components number of pods for m2 and number of grains for pod was significant difference only for the cultivars and plant density. For densities, the higher density of plants resulted in reducing the number of pods for area, grains per pod, among the cultivars NK 70 showed the highest number of pods for m2 and the cultivar BMX Apollo largest number of grains for pod. The mass of 1000 grains was the variable with little variation among worked. There interaction between programs and cultivars, where the maturity cultivars achieved the largest masses. For the participation of thirds in the number of pods, there was only difference among cultivars where the variability of response between the cultivars shows that this variable has genetic domain. / Com o objetivo de avaliar a resposta de dez cultivares de soja submetidas a três arranjos de plantas e à aplicação de oito programas de manejo fungicida no que tange à expressão e ao desenvolvimento da ferrugem asiática, penetração de luminosidade no dossel vegetativo, produtividade e componentes da produtividade da cultura da soja foi realizado experimento durante a safra agrícola 2008/2009, em Itaara/RS. Em campo, as cultivares FCep 55, BMX Apolo, BMX Titan, BRS 243, A 8000,CD 214,NK 70, BRS Pampa, FCep 59 e Monasca foram submetidas às densidades 160, 300 e 440 mil plantas ha-1 e a oito programas de manejo fungicida: P1- testemunha, P2 epoxiconazol +piraclostrobina com aplicação no fechamento da entre linha e 21 dias após a primeira aplicação; P3 epoxiconazol + piraclostrobina aplicado no fechamento da entre linha e 28 dias após; P4 epoxiconazol + piraclostrobina com aplicação em R1 e 21 dias após; P5 epoxiconazol + piraclostrobina com aplicações em fechamento das entre linhas, 21 dias após a primeira aplicação e com terceira aplicação 14 dias após a segunda; P6 tebuconazol + carbendazin com aplicação no fechamento da entre linha e 21 dias após a primeira aplicação; P7 tebuconazol + carbendazin aplicado no fechamento da entre linha e 28 dias após e P8 tebuconazol + carbendazin com aplicação em R1 e 21 dias após. As variáveis avaliadas foram: percentual de penetração de luz no dossel vegetativo, área abaixo da curva de progresso da doença (AACPD), rendimento de grãos e componentes do rendimento. O delineamento experimental utilizado foi o de blocos ao acaso, com 4 repetições, no esquema trifatorial (10 cultivares x 3 densidades de plantas x 8 programas de manejo fungicida). Para a variável percentual de penetração de luz no dossel vegetativo, a cultivar BMX Apolo obteve melhor desempenho com valores próximos a 1% quando submetida à maior densidade. Para as variáveis AACPD e produtividade, as cultivares puderam ser agrupadas como responsivas, não- responsivas e indiferentes à redução da população de plantas. As cultivares responsivas para AACPD foram FCep 55, BMX Apolo, BRS 243, CD 214, já para produtividade tiveram esse comportamento as cultivares FCep 55, A 8000, BMX Titan, BRS 243 e NK 70. Os programas de manejo tiveram a mesma tendência para essas duas variáveis, onde o programa com três aplicações de epoxiconazol + piraclostrobina apresentou menor valor de AACPD e maior rendimento. O número de ramos por planta mostrou que todas as cultivares reduzem a produção de ramos quando expostos a condições adensamento, o que refletiu em concentração da massa de grãos na haste principal, com destaque para a cultivar BMX Apolo que concentrou 99,84% da massa de grãos na haste principal e na maior densidade teve apenas 0,02 ramos por planta. Para os componentes do rendimento número de legumes por m2 e número de grãos por legume, houve diferença significativa somente para os fatores cultivares e densidade de plantas isoladamente. Para as densidades, o adensamento de plantas resultou na redução do número de legumes por unidade de área e de grãos por legume, já entre as cultivares, a cultivar NK 70 apresentou o maior número de legumes por m2 e a cultivar BMX Apolo, o maior número de grãos por legume. A massa de 1000 grãos foi a variável com menor variação dentre as trabalhadas, havendo interação entre os programas e as cultivares, onde as cultivares de ciclo precoce obtiveram as maiores massas. Para a participação dos terços no número de legumes, houve diferença somente entre as cultivares onde a variabilidade de resposta entre os materiais aponta para que essa variável tenha domínio genético.

<p>  </p> <p>Protecting crops from disease requires continuous research because plant pathogen incidence, geographical range, and pathogenicity, are constantly shifting variables as agronomic practices and climate continue to evolve. The objectives of this research are to i.) evaluate field-scale fungicide timing programs for corn (<em>Zea mays L.</em>) diseases at multiple locations in Indiana; ii.) evaluate field-scale fungicide timing programs for soybean (<em>Glycine Max</em> (L.) Merr.) diseases at multiple locations in Indiana; iii.) continue to identify, document, and confirm the distribution of populations of the soybean frogeye leaf spot pathogen (<em>Cercospora sojina)</em> that contain the G143A mutation conferring resistance to quinone outside inhibitor (QoI) fungicides in Indiana; and iv.) assess the incidence, severity, and prevalence of tar spot (<em>Phyllachora maydis</em>) in Indiana. For the first and second objectives, field scale trials were established at three locations in Indiana from 2019 to 2022. No application timings at any location provided significant yield protection for corn or soybeans. To achieve the third objective, 165 isolates of <em>C. Sojina </em>were tested. In total, 24 out of the 32 counties sampled in 2021 and 2022 were documented with QoI-resistance. The fourth objective was accomplished by surveying Indiana counties for incidence and severity of tar spot. As of 2022, 86 out of 92 Indiana counties have been confirmed for the presence of tar spot.</p>

No description available.

Dissertation (PhD(Agric)) -- University of Stellenbosch, 2001. / ENGLISH ABSTRACT: Net blotch of barley, caused by Pyrenophora teres, is one of the most important diseases of this cereal in the south Western Cape Province of South Africa. This fungus exists as two different types (forms), namely a nettype and a spot-type that are distinguished by differential symptom expression on barley leaves. Based on this specific plant pathological difference a series of studies of agricultural importance were executed to investigate the effects of sexual recombination between these two types. In addition, studies were done to determine the difference between local net- and spot-type populations with regards to population structure and fungicide sensitivity. This dissertation therefore, consists of a collection of separate publications and as a result a certain degree of redundancy has been unavoidable. Recombination is one of the most important evolutionary forces involved with sexual reproduction. In plant-fungal agricultural ecosystems this may result in pathogenic fungal populations adapting more rapidly to control programs such as fungicide applications. The first section of the review in part 1 of this dissertation covers different aspects of sexual reproduction in ascomycetes, specifically focussing on mating-type genes, vegetative incompatibility and recombination. The major part of the review is then dedicated to various plant pathological aspects of P.teres, specifically addressing the differences between the two types, and in various cases highlighting the significance of sexual recombination within and between the net- and spot-type. Using morphological criteria for identification purposes there have been many conflicting reports concerning the identity of leaf spot isolates in the Western Cape Province of South Africa. In part 2, the correct identity was eventually achieved employing mating studies and molecular markers .: This was accomplished after single ascospores were obtained from pseudothecia after in vitro mating had occurred between a verified P. teres net-blotch isolate from Denmark and a representative Pyrenophora leaf spot isolate from South Africa. Using amplified fragment length polymorphism (AFLP) and RAPD markers, recombination was demonstrated in the progeny that had DNA banding patterns different from the two parental isolates. Pathogenicity trials also confirmed that recombination had taken place during mating. Inoculations were conducted on the differential cultivars susceptible to the net-blotch and leaf spot forms. The two parents induced typical net-blotch or leaf spot symptoms whereas the progeny mostly induced a jagged spot symptom on each cultivar. Fungicide sensitivity tests using the ergosterol biosynthesis inhibitors showed that, due to recombination, some progeny could have increased resistance to these fungicides. Due to mating and subsequent recombination between a net blotch isolate of P. teres and a representative leaf spot isolate, it was concluded that the latter was P. teres f. maculata. Fifteen of the net-spot hybrid progeny (F1) produced from the mating study in Part 2 were screened in Part 3 to assess their viability and genetic stability. Hybrid progeny (F1) inoculated onto barley seedlings consisting of the cultivars Stirling (differentially susceptible to net-type isolates), B87/14 and Clipper (both differentially susceptible to spot-type isolates) produced intermediate symptoms on all cultivars. Axenic cultures (F1-1) isolated from foliar lesions, followed by repeated inoculation and isolation (F1-2) onto a healthy set of seedlings produced similar intermediate symptoms. RAPDs conducted with two 1Q-mer primers on all isolates of F1-1and F1-2progeny revealed profiles similar to those obtained for F1 isolates. RAPD molecular data, therefore, indicated that hybrid progeny of this net x spot mating were genetically stable after having been subjected to two repetitive inoculation and reisolation cycles. Phylogenetic analysis of DNA sequences of the internal transcribed spacers (ITS1 and ITS2) flanking the 5.8S nuclear ribosomal RNA gene and the 5' end partial histone-3 gene confirmed the genetic stability of the hybrid progeny. These results also indicated that the hybrid progeny produced consistent symptoms throughout the series of experiments, and maintained their virulence to the differential cultivars screened. Both types of P. teres are prevalent in the south Western Cape Province of South Africa, found on susceptible cultivars often grown within close proximity of each other. In Part 4, a net- and spot-type population were characterised in terms of their population structure using RAPD markers. Samples were collected from infected barley leaves from two separate quadrants in each field, the two quadrants positioned in corners of the fields, diagonal to one another. A total of 65 loci were produced of which 54 were polymorphic. Total gene diversities determined for all loci resulted in mean indices of 0.063 and 0.082 being obtained respectively for the net- and spottype populations. A coefficient of genetic differentiation (Gs) of 0.0149 was obtained between sites within populations while a coefficient (GT) of 0.63 was obtained between the two populations. Genotypic variation revealed 13 distinct multilocus genotypes (haplotypes) in the net-type population while there were 12 in the spot-type population. UPGMA cluster analysis done on the two populations together with six progeny from the mating between a netand spot-type isolate resulted in three main clusters being produced, one for each population and one for the progeny. One isolate collected from the nettype population also contained a unique spot-type RAPD fragment. This suggested that sexual recombination may be taking place between isolates of the net- and spot-type under field conditions. Fungicide application is the most important method used in the control of net blotch in South Africa. In Part 5 the fungicide sensitivities (ICsD values) of 89 monoconidial isolates (46 net-type and 43 spot-type) of P. teres to sterol demethylation inhibiting fungicides were determined, based on the inhibitory effect on radial mycelial growth. The fungicides evaluated were triadimenol, bromuconazole, flusilazole, propiconazole and tebuconazole. Both net- and spot-type isolates revealed strong resistance to triadimenol while flusilazole was shown to be the strongest inhibitor of fungal growth. Spot-type isolates showed a higher resistance than net-type isolates to all five fungicides screened. The ICsD values indicated significant differences between four of the fungicides (triadimenol, tebuconazole, flusilazole and propiconazole). The ICsD values between propiconazole and bromuconazole were not significant. This study suggested that spot-type isolates showed a higher degree of resistance to commercially used fungicides than net-type isolates. The overall conclusion of this study is that the spot-type of P. teres is the pathogen associated with leaf spots of barley in the south western Cape province of South Africa and not P. japonica as earlier reported. Together with the net-type, both types exist as genetically variable populations in this barley production region. Mating between the two types results in sexual progeny that are genetically stable. This implies that barley fields adjacent to one another in which either net- or spot-type susceptible cultivars are being cultivated may lead to sexual progeny being produced. This in turn may lead to an increased rate at which fungal populations may become resistant to commercially used fungicides. It is furthermore suggested that an alternative fungicide seed treatment is used instead of triadimenol due to high resistance of P. teres to this fungicide. / AFRIKAANSE OPSOMMING: Netvlek op gars is een van die belangrikste siektes van hierdie graansoort in die suidelike deel van die Westelike Kaapprovinsie. Dié siekte word veroorsaak deur die swam Pyrenophora teres. Hierdie swam kom voor as twee verskillende tipes, naamlik 'n net-tipe en 'n kol-tipe wat onderskei word op grand van die voorkoms van hulle simptome op garsblare. Hierdie planpatologiese verskil in ag genome, is 'n reeks studies van landboukundige waarde uitgevoer om die effek van geslagtelike rekombinasie tussen die twee tipes te ondersoek. Daarbenewens is ook studies uitgevoer om om die verskil te bepaal tussen plaaslike net- en koltipe populasies ten opsigte van populasiestruktuur en fungisiedsensitiwiteit. Hierdie verhandeling bestaan dus uit 'n versameling afsonderlike publikasies en as gevolg daarvan is daar onvermydelik'n mate van oorvleueling. Rekombinasie is een van die belangrikste evolusionêre kragte betrokke by geslagtelike voortplanting. In plant-swam landboukundige ekostelsels kan dit veroorsaak dat patogene swampopulasies vinniger aanpas by beheerpragramme soos fungisiedtoediening. Die eerste gedeelte in deel 1 van hierdie verhandeling dek die verskillende aspekte van geslagtelike voortplanting van ascomycetes, met spesifieke verwysing na paringstipe gene, vegetatiewe onverenigbaarheid en rekombinasie. Die grootste gedeelte van die oorsig word gewyaan verskeie plantpatologiese aspekte van P. teres,en wys veralop die verskille tussen die twee tipes. In verskeie gevalle word die betekenis van geslagsrekombinasie binne en tussen die net- en koltipe uitgelig. Deur morfologiese kenmerke vir identifikasiedoeleindes te gebruik, is daar baie teenstrydige verslae rakende die identifikasie van blaarvlekisolate in die Westlike Kaapprovinsie van Suid-Afrika. In deel 2 is die korrekte identifikasie eventueel verkry deur gebruik te maak van paringstudies en molekulêre merkers. Dit is bereik nadat enkel ascospore verkry is uit pseudothecia gevorm na in vitro paring plaasgevind het tussen 'n bevestigde P. teres netvlek isolaat uit Denemarke en 'n verteenwoordigende Pyrenophora blaarvlekisolaat van Suid- Afrika. Deur gebruik te maak van versterkte fragmentlengte polimorfisme [AFLP] en RAPD merkers, is rekombinasie gedemonstreer in die nasate wat DNA bandpatrone gehad het wat verskil het van dié van die "ouer" isolate. Patogenisiteitstoetse het ook bevestig dat rekombinasie tydens paring plaasgevind het. Inokulasies is uitgevoer op die verskillende cultivars wat vatbaar is vir die netvlek en blaarvlek vorme. Die twee ouers het tipiese netvlek of blaarvlek simptome veroorsaak, terwyl die nasate hoekige vlekke veroorsaak het op elke cultivar. Toetse vir fungisiedsensitiwiteit deur gebruik van die ergosterol biosintese inhibeerders het gewys dat a.g.v. rekombinasie sekere nasate verhoogde weerstand teen hierdie fungisiedes het. As gevolg van paring en daaropvolgende rekombinasie tussen 'n netvlek isolaat van P. teres en 'n verteenwoordigende blaarvlek isolaat is afgelei dat laasgenoemde P. teres f. maculata is. Vyftien van die netvlek hibried nakomelinge (F1) verkry van die paringstudie in deel 2 is ondersoek in deel 3 om hul lewensvatbaarheid en genetiese stabiliteit te bepaal. Hibried nasate (F1) geïnokuleer op garssaailinge bestaande uit die volgende cultivars: Stirling (soms vatbaar vir net-tipe isolate) , B87/14 en Clipper (albei soms vatbaar vir kol-tipe isolate) het intermediêre simptome op al die cultivars veroorsaak. Akseniese kulture (F1-1) geïsoleer uit blaarletsels gevolg deur herhaalde inokulasie en isolasie (F1-2) op 'n gesonde stel saailinge het dieselfde intermediêre simptome veroorsaak. RAPDs uitgevoer met twee 10-mer inleiers op al die isolate van F1-1 en F1-2 nasate het profiele opgelewer soortgelyk aan dié wat vir F1 isolate verkry is. RAPD molekulêre data het dus gewys dat die hibried nasate van hierdie net x kol paring geneties stabiel was nadat dit onderwerp is aan twee inokulasie en reïsolasie siklusse. Genetiese stabiliteit van die hibried nageslag is bevestig deur filogenetiese analise van die DNA volgorde van die interne getranskribeerde spasieerders (ITS1 en ITS2) reg langs die 5.8S nukluêre ribosomale RNA geen en die 5' end gedeeltelike histoon-3 geen. Hierdie resultate het ook gewys dat die hibried nasate konstante simptome getoon het tydens die hele reeks eksperimente en hulle virulensie behou het vir die kultivars wat getoets is. Beide tipes van P. teres kom algemeen voor in die suidelike deel van die Westelike Kaapprovinsie en word gevind op vatbare cultivars wat dikwels naby mekaar groei. In deel 4 is 'n net- en kol-tipe populasie gekarakteriseer in terme van hulle populasiestruktuur deur gebruik van RAPD merkers. Monsters is versamel van geïnfekteerde garsblare van twee aparte kwadrante in elke saailand. Die twee kwadrante is geplaas in die hoeke van die saailand, diagonaal tot mekaar. 'n Totaal van 65 lokusse is gevorm, waarvan 54 polimorfies was. Die algehele genetiese verskeidenheid bepaal vir alle lokusse, het gelei tot gemiddelde indekse van 0.063 en 0.082 soos gevind vir die net- en kol-tipe populasies. 'n Koëffisiënt van genetiese differensiasie (Gs ) van 0.0149 is gevind tussen gebiede tussen populasies, terwyl 'n koëffisiënt (GT) van 0.63 gevind is tussen die twee populasies. Genotipiese variasie het 13 duidelike multilokus genotipes (haplotipes) getoon in die net-tipe populasie, terwyl daar twaalf was in die kol-tipe populasie. UPGMA groeperingsanalises wat gedoen is op die twee populasies tesame met ses nasate van die paring van 'n net- en koltipe isolaat het tot gevolg gehad dat drie hoof groepe gevorm is, een vir elke populasie en een vir die nasate. Een isolaat wat versamel is, van die net-tipe populasie het 'n unieke kol-tipe RAPD fragment bevat. Dit wys daarop dat geslagtelike rekombinasie in veldomstandighede mag voorkom tussen isolate van die net- en kol-tipe. Fungisiedtoediening is die belangrikste metode wat gebruik word om netvlek in Suid-Afrika te beheer. In deel 5 is die fungisiedsensitiwteit (Ieso waardes) van 89 enkelkonidiale isolate (46 net-tipe en 43 kol-tipe) van P. teres teen sterol demetielasie inhiberende fungisiedes bepaal, op die basis van die onderdrukkende effek op die radiale groei van die miselium. Die volgende fungisiedes is geëvalueer: triadimenol, bromuconazole, flusilazole, propiconazole en tebuconazole. Beide net- en kol-tipe isolate het 'n sterk weerstand teen triadimenol openbaar, terwyl flusilazole gevind is as die sterkste onderdrukker van swamgroei. Kol-tipe isolate het 'n hoër weerstand as die net-tipe isolate teen al vyf fungisiedes wat getoets is, gehad. Die lesowaardes het aangedui dat daar beduidende verskille tussen vier van die fungisiedes IS (triadimenol, tebuconazole, flusilazole en propiconazole). Die leso waardes tussen propiconazole en bromuconazole was nie beduidend nie. Die gevolgtrekking van hierdie studie is dus dat die kol-tipe isolate 'n hoër graad van weerstand teen kommersiëel gebruikte fungisiedes as die net-tipe isolate gehad het. Die algehele gevolgtrekking van hierdie studie is dat die kol-tipe van P. teres, die patogeen is wat geassosieer word met blaarvlekke op gars in die suidwestelike Kaapprovinsie van Suid-Afrika, en nie P. japonica soos voorheen gerapporteer nie. Tesame met die net-tipe, kom altwee tipes voor as geneties veranderlike populasies in hierdie gars verbouingstreek. Paring tussen die twee tipes lei tot geslagtelike nasate wat geneties stabiel is. Dit impliseer dat aangrensende garsvelde waarop net- óf kol-tipe vatbare kultivars verbou word, mag lei tot die produksie van geslagtelike nasate. Dit kan weer lei tot 'n verhoogde tempo waarteen swampopulasies weerstandbiedend teenoor kommersiële fungisiedes raak. Daar word verder ook voorgestel dat alternatiewe fungisied saadbehandelings gebruik word in plaas van triadimenol as gevolg van verhoogde weerstand van P. teres teenoor laasgenoemde.

Target spot is a foliar disease of soybean that can produce yield losses. The disease has recently become a concern due to increasing intensity of its epidemics in the main soybean growing countries (USA, Brazil and Argentina). The goals of this study were to i) estimate the target spot control efficiency and yield response of labeled fungicides for the main soybean growing region of Brazil and identify factors affecting their performance; ii) characterize the relationship between target spot severity and soybean yield using meta-analytic techniques, and to identify patterns which allow understanding the heterogeneity in the relationship; iii) to explore the variability of genetic resistance of cultivars and verify the less time-consuming methodology for doing it; iv) explore the pathogen-host interaction effects on the epidemic development; v) compare the sensitiveness of the canopy strata position injured by C. cassiicola to reduce grain yield. Fungicides containing fluxapyroxad + pyraclostrobin were the most efficient ones to control target spot, with control levels of 75% and their yield response depended on the disease pressure (DP, DPLow < 35% target spot severity at untreated checks <= DPHigh). At DPLow was unprofitable the use of fungicides and at DPHigh the latter fungicides had the best performances increasing yield relative to the untreated check in > 469 kg ha-1 (+19.1%). Potential yield of soybean in absence of target spot was estimated in 3507 kg ha-1 for 41 trials in Brazil and the percentual reduction for each target spot severity point was calculated in 0.48%, what would represent a reduction of 24% in a hypothetical target spot severity of 50% (ranging from 8% to 42%). The soybean cultivar had a significant effect to explain this wide range of responses: potential losses of 11%, 18.5% and 42% was calculated for cultivar BMX Potência RR, TMG803 and M9144RR respectively at target spot severity = 50%. We observed high variability on the genetic resistance in the tested germplasm: cultivars with very low target spot intensity (BRS360) to highly susceptible cultivars. A single-point disease severity assessment was not as reliably as an integrative three-point assessment, which had no difference with a less time-consuming two-point disease assessment. In trials where 3 C.cassicola isolates from different regions of Brazil were inoculated individually on 3 soybean cultivars contrasting in their resistance level we observed that the incubation period, disease severity, lesion density and lesion size at 14 days after inoculation, were influenced by the cultivar and not by isolate geographical origin. Using disease severity coupled with defoliation assessments throughout the reproductive growth stages, we performed correlations between grain yield and the leaf area injury at different plant positions - growth stages. The best correlation was observed with the assessments at R5.5 for injuries at middle canopy section. / A mancha alvo é uma doença foliar que pode ocasionar perdas de rendimento na cultura da soja. A doença tornou-se recentemente uma preocupação nos principais países produtores de soja (EUA, Brasil e Argentina), devido à crescente intensidade de suas epidemias. Os objetivos deste estudo foram: i) estimar a eficiência de controle da mancha alvo por fungicidas comerciais registrados no Brasil e a resposta em rendimento da soja, identificando fatores que afetam o desempenho dos fungicidas; ii) caracterizar a relação entre a severidade da mancha alvo e o rendimento da soja, utilizando técnicas meta-analíticas e identificar padrões que permitam compreender a heterogeneidade existente nessa relação; iii) explorar a variabilidade da resistência genética de cultivares de soja e verificar a metodologia mais eficiente para discriminar cultivares suscetíveis de resistentes; iv) explorar os efeitos da interação entre a origem do isolado de C. cassiicola e a cultivar de soja no desenvolvimento da epidemia; v) comparar a sensibilidade da posição do estrato do dossel afetado por C. cassiicola na redução do rendimento de grãos. Fungicidas compostos pelos ingredientes ativos fluxapyroxad + piraclostrobina foram os mais eficientes para controlar a mancha alvo, com níveis de controle de até 75%, e sua resposta ao rendimento dependeu da pressão da doença (PD, PDbaixa < 35% de severidade da mancha alvo na testemunha não tratada <= PDalta). Na PDbaixa, a aplicação de fungicidas não foi lucrativa, e em PDalta os fungicidas compostos por fluxapyroxad + piraclostrobina apresentaram os melhores desempenhos, superando o rendimento em relação à testemunha em 469 kg ha-1 (+ 19,1%). O rendimento potencial geral da soja, na ausência da mancha alvo, foi estimado em 3507 kg ha-1 para 41 ensaios distribuídos no Brasil. A redução no rendimento para cada ponto percentual de incremento na severidade da mancha alvo foi calculada em 0,48%. Com isso, níveis (hipotéticos) de severidade de mancha alvo de 50% ocasionariam uma redução de rendimento de 24% (variando entre 8% a 42%). A cultivar de soja teve um efeito significativo para explicar esta grande amplitude de respostas: reduções potenciais (com 50% de severidade) de 11%, 18,5% e 42% foram calculadas, respectivamente, para as cultivares BMX Potência RR, TMG803 e M9144RR. Foi constatada alta variabilidade na resistência genética no germoplasma testado, desde cultivares com intensidade de mancha alvo muito baixa (BRS360) até cultivares altamente suscetíveis. Avaliações da severidade da doença em uma única data não foi tão confiável quanto a avaliação integrada de duas ou três datas. Em experimentos onde 3 isolados de diferentes regiões do Brasil foram inoculados isoladamente em 3 cultivares contrastantes nos níveis de resistência, observou-se que o período de incubação, a severidade da doença, a densidade de lesões e o tamanho das lesões aos 14 dias após a inoculação foram influenciados pela cultivar. Porém, não foi observado efeito da origem geográfica do isolado de C. cassiicola nos componentes monocíclicos avaliados. A severidade da doença em diferentes posições da planta, juntamente com as avaliações de desfolha ao longo dos estádios de crescimento reprodutivo da soja, foi correlacionada ao rendimento de grãos. A melhor correlação entre o rendimento de grãos e a intensidade da doença (severidade + desfolha) foi observada nas avaliações em R5.5 na seção média do dossel.