A cultura do meloeiro é de grande importância econômica no Brasil, sendo a região Nordeste a principal produtora e exportadora do fruto. A baixa resistência aos principais patógenos é um dos fatores limitantes para a competitividade brasileira da cultura no mercado internacional. Uma das doenças mais importantes é o oídio, causado por Podosphaera xanthii. Geralmente, o controle de P. xanthii é obtido com o uso de fungicidas. Porém, é necessário empregar medidas mais econômicas e que não agridam o ambiente, como a utilização de cultivares resistentes. O objetivo deste trabalho foi identificar marcadores moleculares de vários tipos ligados ao gene Pm-1 de resistência à raça 1 de P. xanthii com o intuito de auxiliar programas de melhoramento genético. Para tanto foram utilizadas duas linhagens quase-isogênicas (LQI) de melão AF426pm1 (P1) e AF426Pm1 (P2), ambas pertencentes à variedade inodorus, que são contrastantes para ausência e presença, respectivamente, do gene Pm-1. Para a análise de co-segregação entre gene e marcadores candidatos, foi utilizada uma população de retrocruzamento RC1F1, fenotipada para resistência a raça 1 de oídio, obtida a partir do cruzamento entre linhagens. As técnicas de LM-PCR e AFLP resultaram em marcadores polimórficos, porém somente os encontrados com a técnica de AFLP puderam ser utilizados como marcadores no teste de co-segregação. Foram encontrados dois marcadores AFLP. No entanto, somente um, denominado HF155 mostrou-se ligado a uma distância de 4,9 cM do gene Pm-1. Devido à proximidade deste marcador ao gene este pode ser utilizado em programas de seleção assistida por marcadores, visando o melhoramento de linhagens com resistência ao P. xanthii. / Melon crop is of great economical importance in Brazil being the Northeast region the main producer and exporter of the fruit. The low levels of resistance to pathogens is one of the restricting factors for the Brazilian competitiveness worldwide. One of the most important diseases is powdery mildew caused by Podosphaera xanthii. Generally, the control of P. xanthii is achieved with the use of fungicides. However it is necessary to use less expensive control methods with a minimum environmental impact such as resistant cultivars. The objective of this work was to identify molecular markers linked to the Pm-1 gene which confers resistance to race 1 of P. xanthii with the purpose of assisting boding program. Two near isogenic lines (LQIs) of melon Agro AF426pm1 (P1) and AF426Pm1 (P2), both belonging to the inodorus variety, which are respectivally contrasting for abscence and presence of Pm-1 gene were analyzed. For the cosegregation analyses between gene and candidate markers, it was used the BC1F1 population was used screened for resistance to powdery mildew and obtained from a cross between the LQIs. The LM-PCR and AFLP techniques were efficient in the polymorphisms detection, however only those ones which were found using AFLP technique could be used as markers in the co-segregation test. It was found two markers with this technique but just the HF155 is located 4.9 cM of the Pm-1 gene. Due the proximity of the HF155 marker to the Pm-1 gene this one can be used in markerassisted selection aiming to develop melon cultivars resistant to P. xanthii.

O meloeiro é uma cultura de relevância econômica no Brasil, principalmente na região Nordeste, onde grande parte da produção é exportada. Entre os fatores limitantes da cultura está o oídio, que pode ser controlado através do uso de variedades resistentes. No entanto, para que isto seja possível, é necessário que se tenha conhecimento sobre qual é o agente causal em ocorrência na região de produção e quais são as fontes de inóculo desta doença. O presente trabalho teve como objetivos o levantamento das espécies e raças de oídio com ênfase na região Nordeste, a verificação da patogenicidade de isolados provenientes de outras cucurbitáceas em relação ao meloeiro, a avaliação de germoplasma de meloeiro para resistência a P. xanthii raça 2 (francesa), a proposição de um sistema de cultivares de meloeiro para caracterização de raças de P. xanthii e a otimização de um protocolo de extração de DNA de P. xanthii. No total de 65 isolados analisados, observou-se que Podosphaera xanthii foi a única espécie encontrada entre as amostras. Dentre estas, as raças 1 e 2 foram as prevalentes. Também foram observadas as raças 0, 3, 4 e 5 em menor freqüência. As culturas de abóbora, abobrinha, melancia e pepino apresentaram-se como fontes de inóculo de oídio para o meloeiro, uma vez que todos os isolados provenientes destas culturas colonizaram discos foliares de meloeiro. Em geral, os genótipos de melão cultivados comercialmente no Brasil apresentaram-se resistentes a raça 2 (francesa) de P. xanthii. A partir da observação das raças ocorrentes no país e de dados da literatura, propôs-se um sistema de cultivares de meloeiro diferenciadoras para raças de P. xanthii a ser empregado em monitoramentos do patógeno. Um protocolo para extração de DNA de P. xanthii também foi obtido com a finalidade de facilitar estudos genéticos sobre o patógeno com base em marcadores moleculares. / Melon is an economically important crop in Brazil, mainly in the north-east region, where most of the production is exported. Among the limiting factors of the crop is powdery mildew, which can be controled by using resistant varieties. However, to make it possible, it is necessary to know the causal agent in occurrence in the production region and what are the inoculum sources of this disease. The present work had as objectives the survey of the powdery mildew species and races with emphasis in the north-east region; the verification of the strains pathogenicity deriving from others cucurbits in melon; the evaluation of melon germplasm to Podosphaera xanthii race 2 (french) resistance; the proposition of a system of melon cultivars to characterize the P. xanthii races and the optimization of a protocol to P. xanthii DNA extraction. Over all of the 65 strains analysed, it was observed that P. xanthii was the unique specie encountered between the samples. Within that, the races 1 and 2 were prevalents. Also were observed the races 0, 3, 4 and 5 at low frequency. Squash, zucchini, watermelon and cucumber crops showed up like inoculum sources of powdery mildew to melon. Usually, the melon genotypes commercially cultivated in Brazil showed up resistant to P. xanthii race 2 (french). After the observation of races in occurrence and datas from literature, a system of melon cultivars was proposed to differentiate P. xanthii races to be used in surveys of the pathogen. A protocol to extraction P. xanthii DNA also was proposed with the purpose to facility genetics studies about the pathogen basis on molecular markers.

A viticultura no Brasil e no Estado de São Paulo encontra-se em expansão nos últimos 10 anos e a cv. Niagara Rosada (Vitis labrusca) se destaca como principal cultivar para produção de uvas para mesa. Essa cultivar é suscetível à várias doenças foliares, como a ferrugem (Phakopsora euvitis) e o míldio (Plasmopara viticola). Não existem estimativas quantitativas dos danos causados por essas doenças na cv. Niagara Rosada. Diante do exposto os objetivos desse trabalho foram quantificar os efeitos da ferrugem e do míldio nas trocas gasosas, nas limitações da fotossíntese, no acúmulo de biomassa, no acúmulo de carboidratos, e na produção da cv. Niagara Rosada e desenvolver um modelo de simulação para V. labrusca acoplado com os mecanismos de danos da ferrugem e míldio. Experimentos foram conduzidos, separadamente para cada doença, em condições controladas, em mudas inoculadas com diferentes concentrações de P. euvitis e P. viticola e em campo experimental. Medidas de trocas gasosas e curvas de resposta da taxa líquida de assimilação de CO2 ao aumento da concentração intercelular de CO2 (Ci) foram realizadas em mudas sadias e infectadas com P. euvitis e P. viticola. Teores de açúcares solúveis totais, sacarose e amido foram determinados em plantas sadias e inoculadas com os patógenos. P. euvitis e P. viticola reduziram a taxa fotossintética em plantas infectadas tanto na área da lesão como no tecido verde adjacente a lesão (lesão virtual). Valores do parâmetro β, indicativo da lesão virtual, foram de 5,7 e 2,9, respectivamente para P. euvitis e P. viticola. P. euvitis reduziu em 48%, 36% e 67% a atividade da Rubisco (Vcmax), a taxa máxima de transporte de elétrons usados para a regeneraração da RuBP (Jmax) e a condutância do mesofilo (gm), respectivamente, em folhas infectadas. A área foliar e biomassa de raízes em mudas inoculadas com P. euvitis foram reduzidas. Através de análises histopatológicas e da quantificação de amido nas folhas sadias e doentes foi possível observar o acúmulo de amido em regiões adjacentes às pústulas de P. euvitis. Mesmo em baixas severidades da ferrugem já se observam reduções no acúmulo de carboidratos em raízes. P. viticola reduziu Vcmax em 23,5 % em folhas infectadas comparadas às folhas sadias. P. viticola reduziu a biomassa de raízes e quantidade de carboidratos nas raízes de mudas doentes e de plantas no campo em ano de alta severidade da doença. Plantas no campo com sintomas míldio produziram em média 0,5 kg a menos do que plantas sadias e a principal causa da redução na produção foi a queda de bagas causada pela infecção de P. viticola nos cachos. Um modelo de simulação para Vitis labrusca foi desenvolvido, utilizando o software Stella®. A produção, partição e dinâmica da biomassa da videira foi simulada ao longo de 20 anos e os efeitos de P. euvitis e P. viticola na redução do tecido verde sadio da planta, na redução da eficiência fotossintética, no desvio de assimilados, na aceleração da senescência foliar e na queda de frutos foram inseridos no modelo. / Viticulture in Brazil and in the state of São Paulo has increased in the last 10 years and cv. Niagara Rosada (Vitis labrusca) is the main cultivar for the production of table grapes. This cultivar is susceptible to several foliar diseases, such as rust (Phakopsora euvitis) and downy mildew (Plasmopara viticola). There are no quantitative estimates of the damage caused by these pathogens in cv. Niagara Rosada. The objectives of this work were to quantify the effects of rust and downy mildew on gas exchange, photosynthesis limitations, biomass accumulation, carbohydrate accumulation and production of cv. Niagara Rosada and to develop a simulation model for V. labrusca coupled with damage mechanisms caused by rust and mildew. Experiments were conducted, independently for each disease, under controlled conditions in potted plants inoculated with different concentrations of P. euvitis and P. viticola and in experimental field, with natural occurrence of diseases. Measurements of gas exchange and response curves of photosynthetic rate to the increase of the intercellular CO2 concentration (Ci) were carried out in healthy plants and infected plants with P. euvitis and P. viticola. Total soluble sugars, sucrose and starch contents were determined in healthy plants and inoculated plants with the pathogens and kept under controlled conditions. P. euvitis and P. viticola reduced the photosynthetic rate in infected plants both in the area of the lesion and in the green tissue adjacent to the lesion (virtual lesion). Values of parameter β, indicative of the virtual lesion, were 5.8 and 2.9, respectively for P. euvitis and P. viticola. P. euvitis reduced the activity of Rubisco (Vcmax), rate of electrons transport contributing for the RuBP-regeneration (Jmax) and the conductance of mesophyll (gm), respectively, on infected leaves by 48%, 36% and 67%. The severity of P. euvitis reduced leaf area and biomass of plant roots. The histopathological analysis and starch quantification in the leaves allowed to observe starch accumulation in regions adjacent to the pustules of P. euvitis. Even in low disease severities, reductions in carbohydrates accumulation of in roots are already observed. P. viticola reduced Vcmax by 23.5% in infected leaves compared to healthy leaves. P. viticola reduced the root biomass and carbohydrate amounts in the roots of potted plants and plants in the field in the year of high disease severity. Field plants with mildew symptoms produced on average 0.5 kg less than healthy plantsand the main cause of reduction in production was the drop of berries caused by infection of P. viticola in the clusters. A simulation model for Vitis labrusca was developed using Stella ® software. The production, partitioning and dynamics of grapevine biomass was simulated over 20 years and the effects of P. euvitis and P. viticola on the reduction of healthy green plant tissue and photosynthetic efficiency, the assimilative sapper, leaf senescence acceleration and fruit drop were coupled in the model.

Les infections multiples sont courantes dans la nature et sont considérées comme très importantes dans l'évolution des caractéristiques biologiques des parasites. Théoriquement, les infections multiples devraient entraîner une évolution de la virulence à la fois comme stratégie adaptative et comme stratégie plastique. Dans cette thèse, j'utilise Hyaloperonospora arabidopsidis, un parasite naturel d'Arabidopsis thaliana, qui s'est avéré pratique pour des études en écologie évolutive, pour étudier: i) les infections multiples consécutives à la co-inoculation et à l'inoculation séquentielle; succès de l'infection et succès de transmission d’une souche individuelle (génotypage par PCR) et des phénotypes d'infection, y compris virulence entre inoculation unique et mixte, iii) effet du délai d'inoculation et d'ordre des souches inoculées sur les phénotypes d'infection et le succès de l'infection individuelle. Ici, j'ai trouvé une fréquence plus élevée de co-infection à la suite de l'inoculation séquentielle que à la suite de la co-inoculation des mêmes combinaisons de souches. L'inoculation mixte de certaines combinaisons de souches a entraîné une modification des phénotypes d'infection, souvent avec un succès d'infection plus faible chez certaines souches à la suite des inoculations en mélanges qu’en inoculation simple. Ce résultat implique une interférence entre les souches dans l'inoculum mixte. La virulence globale de l'infection après l'inoculation mixte n'était pas toujours supérieure à celle de l'infection à souche simple. De plus, les souches uniques utilisées dans ces expériences ne différaient pas toujours les unes des autres en termes de virulence. Le seul test d'un mélange de génotypes à trois souches a provoqué une virulence globale plus élevée que les trois infections à souche unique respectives. Une plus grande virulence globale dans ce cas pourrait être due à la plasticité des souches parasitaires inoculées, à la réponse à la présence d'autres souches dans l'inoculum mixte ou à l'effet de multiples souches supprimant le système de défense de l'hôte. Lorsque les souches ont été inoculées de manière séquentielle et non ensemble, le succès de l'infection de souches individuelles différait entre les différents ordres d'inoculation, ce qui pourrait être dû à des effets indirects via le système de défense de l'hôte. En résumé, l'inoculation séquentielle a semblé réduire l'interférence entre les souches parasitaires, avec un effet de décalage temporel et d'ordre de la souche inoculée sur le succès de l'infection de souches individuelles. Une interférence dans un inoculum mixte peut générer différents succès d'infection et phénotypes d'infection à partir des inoculations individuelles respectives. J'ai trouvé un cas évident de virulence globale plus élevée dans les infections causées par des inoculations mixtes. Par conséquent, une virulence globale plus élevée peut se produire malgré le fait que nous ne trouvions pas de meilleures performances de génotypes plus virulents dans des infections à la suite d'inoculations mixtes. Ainsi, ces résultats ne permettent pas de prédire l’évolution de la virulence supérieure parmi ces combinaisons de souches testées. Cependant, la plasticité des phénotypes des souches inoculées dans l'inoculum mixte a généré une virulence globale de l'infection plus élevée. Ces résultats peuvent aider à comprendre comment les génotypes de parasites répondent aux infections mixtes. / Multiple infections are common in nature, and are considered very important in the evolution of parasite life-history traits. Theoretically, multiple infections should lead to evolution of higher levels of virulence both as an adaptive and as a plastic strategy. In this thesis I use Hyaloperonospora arabidopsidis, a natural parasite of Arabidopsis thaliana, which has proven a useful tool for unlocking some evolutionary ecology questions, to investigate: i) multiple infections following co-inoculation and sequential inoculation, ii) number of infected plants, infection success and transmission success of individual strain (genotyping via PCR), and infection phenotypes including virulence between after single- and mixed inoculation, iii) effect of time lag of inoculation and order of inoculated strain on infection phenotypes and individual strain infection success. Here I found that sequential inoculation contributed higher frequency of co-infection than co-inoculation of the same strain combinations. Mixed inoculum of some strain combinations led to modification of overall infection phenotypes, often with poorer infection success of individual strains compared with that of the more infectious strains. This result implies interference between strains in mixed inoculum. Overall virulence of infection after mixed inoculation was not always higher than that of single strain infection. Furthermore the single strains used in these experiments did not always differ from each other in virulence. The one test of a three-strain mixture of genotypes caused higher overall virulence than the three respective single strain infections. Higher overall virulence in this case might be caused by plasticity of inoculated parasite strains reponse to the presence of other strains in mixed inoculum or an effect of multiple strains suppressing the host defence system. When strains were inoculated sequentially instead of together, infection success of individual strains differed between different orders of inoculation, which could be due to indirect effects via the host defence system. In summary, sequential inoculation seemed to reduce interference between parasite strains, with effect of time lag and order of inoculated strain on infection success of individual strains. Interference in mixed inoculum can generate different infection successs and infection phenotypes from the respective single inoculations. I found one clear case of higher overall virulence in infections caused by mixed inoculations. Thus higher overall virulence can occur despite our not finding higher performance of more virulent genotypes from infections following mixed inoculations. Thus these finding do not predict the evolution of higher virulence among these strain combinations tested. However, plasticity of phenotypes of inoculated strains in mixed inoculum did generate higher overall virulence of infection. These findings can help to understand how the parasite genotypes respond to in mixed infections.

Powdery mildew, caused by the pathogen Blumeria graminis (DC) Speer (Syn. Erysiphe graminis DC) f. sp. tritici, is a major disease of wheat (Triticum aestivum L.). Race-specific resistance is easily identified in the field due to its qualitative phenotype and it is easy to incorporate because it is inherited as a single gene. Unfortunately, this type of resistance is easily overcome by the pathogen. Traits associated with quantitative trait loci (QTL) such as adult-plant resistance (APR), have become popular with plant breeders because of their durability over a wide geographic range and time. Due to the quantitative nature of these genes, they are difficult to study requiring multiple assessments of disease development under natural conditions in more than one location over a period of several weeks. Numerous QTL for APR to powdery mildew have been mapped in independent studies in different wheat backgrounds. The wheat cultivar Massey has been the subject of several studies due to its APR to powdery mildew that has remained effective for several decades. However, it has been difficult to identity simple sequence repeat (SSR) markers that are tightly linked to the QTL for APR in Massey. Such markers give breeders an advantage by allowing them to quickly identify and select for traits that would be difficult to distinguish in the field among breeding progeny from several backgrounds. Therefore, identification of tightly linked markers associated with APR to powdery mildew is necessary so that these traits can be selected for reliably in progeny. / Master of Science

Downy mildew has been a potentially devastating constraint to crop production within the Brassicaceae family worldwide (CABI, 2022) and on the Central Coast of California (Smukler, et al, 2008; Koike, et al. 2007). Downy mildew is a foliar disease, caused by the pathogens in the Hyaloperonospora genus on plants within the Brassicaceae family (Goker, et al, 2009). Downy mildew is a disease that is common in region of the world with Coastal climates that provide humidity and moderate temperatures (Sarahan, et al, 2017). Crops within the Brassicaceae family maintain a predominant role in agricultural systems worldwide (Cartea, et al, 2011) and on the Central Coast of California, in Monterey, San Luis Obispo, Santa Cruz, San Benito, and Ventura Counties (Hidalgo, et al, 2022; Settevendemie, et al, 2022; Sanford, et al, 2022; Griffin, et al, 2022; Chang, et al, 2022; Williams, et al, 2022). The crop production within the Brassicaceae family includes broccoli, cabbage, and cauliflower, and the spring mix production of wild arugula, cultivated arugula, and baby kale. In the first study, an in-lab screening of wild arugula (Diplotaxis tenuifolia) for downy mildew resistance was conducted to identify accessions that contain potential resources of resistance to Hyaloperonospora diplotaxidis. The germplasm of 199 wild arugula accessions was grown out under greenhouse conditions and inoculated with a v single isolate of H. diplotaxidis. A subset of 40 accessions of wild arugula were further inoculated with an additional three isolates of H. diplotaxidis and the ten most resistant accessions were selected for further inoculations. The inoculations of the final subset of ten accessions with the same three isolates were replicated an additional two times. When a factorial ANOVA was conducted to test the interaction between the isolate and accession used, it was found that the impact of the accession on disease incidence was independent from the isolate (P=0.993). Three accessions, 185, 79, and 17 were identified as a potential source of resistance to the isolates utilized in this study, for maintaining a disease incidence below 1%. To compare the results found in the in-lab resistance screening, a field trial was conducted with a subset of wild arugula accessions with diverse reactions to downy mildew. When a regression was conducted comparing the disease incidence under field and lab conditions, the R2 value was found to be 0.37. A second study was conducted to characterize the genetic, morphological, and host range diversity of downy mildew causing pathogens in the Hyaloperonospora genus of hosts in the Brassicaceae family. An isolate collection of approximately 40 isolates of Hyaloperonospora spp. was established and the phylogenetic relationships of these isolates were analyzed by creating two phylogenetic trees using their cox2 mtDNA and ITS rDNA sequences. The H. brassicae and H. diplotaxidis isolates from this study formed separate monophyletic groups in both the cox2 mtDNA and ITS rDNA phylogenetic analyses. When the host range of H. brassicae and H. diplotaxidis were characterized by inoculating a genetically and morphologically diverse set of four host plant species, the H. brassicae and H. diplotaxidis were found to have distinct host ranges. The H. brassicae isolate was found to be parasitic of all B. oleracea morphotypes including broccoli, cabbage, kale, kohlrabi, and ornamental cabbage. The H. diplotaxidis isolate was found to only be parasitic on wild arugula, not cultivated arugula, wild mustard, or any of the B. oleracea morphotypes. The difference between sporangia of H. brassicae and H. diplotaxidis were found to be statistically significant, where the sporangia of H. brassicae were nearly circular and the sporangia of H. diplotaxidis were closer to an oval.

The use of biological control agents (BCAs) in the past has shown limited success as its application has often been done incorrectly, and in addition, management practices are rarely altered to incorporate BCAs. Criteria for the correct application of BCAs have been devised as part of the research, and companies selling these products may use the said criteria. Such application will ensure the correct BCAs are used and, more specifically, used under the correct conditions. The powdery mildew (PM) fungus is often seen to develop resistance to synthetic fungicides and, therefore, alternative control measures are required. BCAs as an alternative pose less risk to the environment, workers and the consumer. A pre-trial has been conducted with a range of BCAs to see if they can control powdery mildew (PM) in a greenhouse environment on hydroponically grown cucumber (Cucumis sativus L.) plants using the variety Baccara that has only a moderate tolerance to PM. The BCAs have been compared to the control (synthetic fungicide: Bravo). Comparative work includes Coyier's model, which has been modified and adapted for these trials to determine the percentage of leaf area covered by the PM infection. Furthermore, the number of fruit harvested per treatment, kilogram yield, total mass of yield and average fruit mass is also used to determine the efficacy of the BCAs as these factors have economic significance to commercial growers. The pre-trial showed promise until the fertigation computer failed, resulting in a nutrient shortage and imbalance, confirming that BCAs alone cannot control PM. Synthetic fungicides were applied until control of PM and plant nutrition was regained. BCAs were re-introduced and used until the end of crop production. The confirmation from the pre-trial has led to the inclusion of silicon in conjunction with the BCAs in the two subsequent trials (Trials 1 & 2). Silicon was applied with the BCAs as a foliar spray on a weekly basis. In trials 1 and 2, the cucumber variety, Palladium, with a high genetic tolerance to PM is used, as this variety is suited to form part of the holistic approach used for trials 1 and 2. Trial 1 showed that treatment A, containing Streptomyces griseovirdis and Streptomyces aureofaciens, had the highest yield. Both of these are bacterial BCAs and demonstrated their adaptability to varied climatic conditions, notably when low humidity was experienced. In treatment B, Trichoderma harzianum strains, Rifai and Uppington, show the slowest rate of PM development. In trials 1 and 2, the best actual PM control was obtained by two fungal based BCAs (Trial 1, treatment C was Ampelomyces quisqualis) and (Trial 2, treatment B was Trichoderma harzianum strains, Rifai and Uppington), showing that fungal BCAs have a place for this application, but the growth-enhancing properties of bacterial based BCAs make economic sense and would make them attractive to growers. Treatment A (Streptomyces spp.) had the most number of fruit for the entire growing period and the best overall yield (kg yield) again. Two of the BCA / silicon treatments have marginally better PM control compared to that of the control (E) treatment, although not statistically significant. Treatment E (control) has the highest average fruit mass in this instance but does not have the highest yield (kg yield) when compared to treatments A and B, possibly due to the growth-enhancing properties of most of these BCAs. Therefore, most of these BCA treatments give fairly inconsistent results that vary possibly according to season, humidity and temperature, making it difficult to predict their efficacy. Using combinations or weekly alternations of these BCAs with extremes of climatic adaptation will probably be the most reliable method of obtaining consistent results. Bacterial BCAs are shown to have lower humidity requirements and produce the most consistent results in terms of fruit number, yield and fruit mass and a combination of bacterial and fungal based BCAs would possibly be the best as this would control PM and yet still have the growth enhancing properties from the bacterial based BCAs. From the research, it can be said that some BCAs in trials 1 and 2 produce results similar to that of the control in terms of percentage leaf area covered by PM and some are shown to have improved yields. Results produced from certain BCA treatments are thus equal to the control; yet provide an environmentally friendly alternative to synthetic fungicides. Silicon is listed as a beneficial element rather than an essential element; however, literature claims it to be highly effective in treating PM in cucurbits. Results from trials 1 and 2 show that control of PM is possible in most cases, when a holistic approach is used. This approach includes a cucumber variety with a high PM tolerance, optimum nutrition, cultural practices and silicon in combination with the BCAs. A complete change of management practices is necessary to implement such a BCA program. / Agriculture, Animal Health & Human Ecology / M. Tech. (Nature Conservation)

The use of biological control agents (BCAs) in the past has shown limited success as its application has often been done incorrectly, and in addition, management practices are rarely altered to incorporate BCAs. Criteria for the correct application of BCAs have been devised as part of the research, and companies selling these products may use the said criteria. Such application will ensure the correct BCAs are used and, more specifically, used under the correct conditions. The powdery mildew (PM) fungus is often seen to develop resistance to synthetic fungicides and, therefore, alternative control measures are required. BCAs as an alternative pose less risk to the environment, workers and the consumer. A pre-trial has been conducted with a range of BCAs to see if they can control powdery mildew (PM) in a greenhouse environment on hydroponically grown cucumber (Cucumis sativus L.) plants using the variety Baccara that has only a moderate tolerance to PM. The BCAs have been compared to the control (synthetic fungicide: Bravo). Comparative work includes Coyier's model, which has been modified and adapted for these trials to determine the percentage of leaf area covered by the PM infection. Furthermore, the number of fruit harvested per treatment, kilogram yield, total mass of yield and average fruit mass is also used to determine the efficacy of the BCAs as these factors have economic significance to commercial growers. The pre-trial showed promise until the fertigation computer failed, resulting in a nutrient shortage and imbalance, confirming that BCAs alone cannot control PM. Synthetic fungicides were applied until control of PM and plant nutrition was regained. BCAs were re-introduced and used until the end of crop production. The confirmation from the pre-trial has led to the inclusion of silicon in conjunction with the BCAs in the two subsequent trials (Trials 1 & 2). Silicon was applied with the BCAs as a foliar spray on a weekly basis. In trials 1 and 2, the cucumber variety, Palladium, with a high genetic tolerance to PM is used, as this variety is suited to form part of the holistic approach used for trials 1 and 2. Trial 1 showed that treatment A, containing Streptomyces griseovirdis and Streptomyces aureofaciens, had the highest yield. Both of these are bacterial BCAs and demonstrated their adaptability to varied climatic conditions, notably when low humidity was experienced. In treatment B, Trichoderma harzianum strains, Rifai and Uppington, show the slowest rate of PM development. In trials 1 and 2, the best actual PM control was obtained by two fungal based BCAs (Trial 1, treatment C was Ampelomyces quisqualis) and (Trial 2, treatment B was Trichoderma harzianum strains, Rifai and Uppington), showing that fungal BCAs have a place for this application, but the growth-enhancing properties of bacterial based BCAs make economic sense and would make them attractive to growers. Treatment A (Streptomyces spp.) had the most number of fruit for the entire growing period and the best overall yield (kg yield) again. Two of the BCA / silicon treatments have marginally better PM control compared to that of the control (E) treatment, although not statistically significant. Treatment E (control) has the highest average fruit mass in this instance but does not have the highest yield (kg yield) when compared to treatments A and B, possibly due to the growth-enhancing properties of most of these BCAs. Therefore, most of these BCA treatments give fairly inconsistent results that vary possibly according to season, humidity and temperature, making it difficult to predict their efficacy. Using combinations or weekly alternations of these BCAs with extremes of climatic adaptation will probably be the most reliable method of obtaining consistent results. Bacterial BCAs are shown to have lower humidity requirements and produce the most consistent results in terms of fruit number, yield and fruit mass and a combination of bacterial and fungal based BCAs would possibly be the best as this would control PM and yet still have the growth enhancing properties from the bacterial based BCAs. From the research, it can be said that some BCAs in trials 1 and 2 produce results similar to that of the control in terms of percentage leaf area covered by PM and some are shown to have improved yields. Results produced from certain BCA treatments are thus equal to the control; yet provide an environmentally friendly alternative to synthetic fungicides. Silicon is listed as a beneficial element rather than an essential element; however, literature claims it to be highly effective in treating PM in cucurbits. Results from trials 1 and 2 show that control of PM is possible in most cases, when a holistic approach is used. This approach includes a cucumber variety with a high PM tolerance, optimum nutrition, cultural practices and silicon in combination with the BCAs. A complete change of management practices is necessary to implement such a BCA program. / Agriculture, Animal Health and Human Ecology / M. Tech. (Nature Conservation)

L’oïdium est une des maladies les plus communes sur chêne en Europe. Cette maladie a été signalée à partir du début du 20ème siècle. Toutefois, peu de données sont disponibles sur l’identité de l’agent causal et de son interaction avec l’hôte. L’objectif de cette étude est d’améliorer les connaissances sur ce pathosystème : (1) caractériser son agent causal avec un marqueur moléculaire (ADN ribosomal) et étudier la distribution spatiale des différentes lignées ; (2) étudier l’adaptation locale du parasite à son hôte ; (3) explorer le déterminisme génétique de la résistance de l’hôte. L’étude de l’ADN ribosomal (ITS=Internal transcribed region et IGS= Intergenic spacer) a mis en évidence l’existence de quatre haplotypes, dont trois seulement avaient été associés à des agents d’oïdium sur chêne en Europe. Erysiphe alphitoïdes, classiquement considéré comme responsable de l’épidémie du XXème siècle est bien l’espèce prépondérante (environ 80% des détections), E. hypophylla et Phyllactinia sp étant détectés à fréquence beaucoup plus faible. Un résultat inattendu est la détection d’une quatrième séquence, présentant 100% d’homologie avec celle de plusieurs agents d’oïdium d’arbres tropicaux et avec Erysiphe quercicola, très récemment décrit sur chêne en Asie et Australie mais jusqu’alors inconnu en Europe. L’ITS de E. quercicola est détecté dans toutes les régions de France, à une fréquence non négligeable (de l’ordre de 15% en moyenne). On le trouve dans les mêmes parcelles et parfois sur les mêmes arbres, voire dans les mêmes lésions que E. alphitoides. La mise au point d’une méthode d’inoculation artificielle a permis d’étudier de façon quantitative l’interaction chêne-oïdium. Une expérimentation d’inoculations réciproques montre que les souches d’oïdium sont plus performantes sur les descendants des arbres dont elles ont été isolées que sur ceux d’autres arbres, suggèrant une adaptation trans- générationnelle. Des inoculations artificielles, complétées par des observations en conditions naturelles, ont également permis de démontrer un déterminisme génétique de la résistance des chênes, avec l’identification de plusieurs QTL. Certains de ces QTL co-localisent avec des QTL de phénologie, en accord avec l’importance de la résistance ontogénique des chênes à l’oïdium. L’ensemble de l’étude amène à reconsidérer l’oïdium du chêne comme un probable complexe d’espèces cryptiques, dont l’histoire de l’invasion et la co-existence en Europe restent à préciser. Les résultats acquis sur l’héritabilité de la résistance et l’adaptation locale du parasite constituent une première étape dans la compréhension des interactions démo-génétiques entre hôte et parasite dans ce pathosystème. / Powdery mildew is the most common disease on oaks in Europe where it was first recorded at the beginning of the 20th century. Yet, little is known about the identity of the causal agent and his interaction with its host. The objective of this study was: (1) to characterize the species responsible of oak powdery mildew with a molecular marker (ribosomal DNA); to study the spatial distribution of these different lineages and (2) to study local adaptation of the parasite to its host (3) to explore the genetic determinism of host resistance. The study of the ITS (internal transcribed region) and IGS (intergenic spacer) diversity revealed the existence of four haplotypes, only three had been already associated with oak powdery mildew in Europe. Erysiphe alphitoïdes, for a long time considered as sole responsible for the epidemic of the XXth century, was the predominant species (~ 80% of detections), E. hypophylla and Phyllactinia sp. were detected at lower frequencies. An unexpected result is the detection of a fourth sequence which show 100% homology with ITS sequences of several powdery mildew agents of tropical trees and Erysiphe quercicola, recently described on oak in Asia and Australia but previously unknown in Europe. E. quercicola ITS was detected in all French regions, at a significant frequency (~ 15%). The study showed that E. alphitoides was often found in association with different ITS types in the same region, the same tree, and even in the same lesion. The development of an artificial inoculation method allowed the quantitative evaluation of the oak-powdery mildew interaction. A reciprocal inoculation experiment showed that powdery mildew strains were more efficient on their mother tress and their descendants than on the other trees, suggesting a trans-generational adaptation. Artificial inoculations, supplemented by observations in natural conditions, have also demonstrated a genetic determination of resistance of oak trees, with the identification of several QTL. Some of these QTL co-localize with QTL controlling phenology, in agreement with the importance of oaks ontogenic resistance to powdery mildew. The entire study leads to reconsider oak powdery mildew as a probable complex of cryptic species; the invasion history and the co-existence in Europe are still to be determined. The results achieved on the heritability of resistance and localization of the parasite are a first step in understanding the demo-genetic interactions between host and parasite in this pathosystem.

A região noroeste do estado de São Paulo é um importante pólo produtor de uvas de mesa, porém possui condições ambientais muito propícias à ocorrência de doenças fúngicas durante todo o ciclo da videira. Alternativas como o uso de coberturas plásticas e de sistemas de alerta fitossanitário têm se mostrado bastante vantajosas para tal, porém, ainda com poucos estudos sobre isso na região. Deste modo, objetivou-se com este estudo avaliar a eficácia de sistemas de alerta fitossanitário no controle do míldio (Plasmopara viticola) em videiras cultivadas sob coberturas plásticas, e, consequentemente, na produtividade e na qualidade das uvas, no Noroeste Paulista. O experimento foi realizado na Estação Experimental de Viticultura Tropical (EVT) da Embrapa Uva e Vinho, localizada no município de Jales, SP, durante os anos de 2012 e 2013. Foram conduzidas três ruas de 120 m de videiras, cultivar apirênica \'BRS Morena\', em espaçamento de 3,0 m entre plantas. Metade do vinhedo foi coberto com filme plástico de polipropileno trançado sobre estrutura metálica em forma de arco (PPT) e a outra metade com tela preta, com 18% de sombreamento (TP18%). O delineamento experimental foi o de blocos casualizados compostos por cinco tratamentos, com seis repetições por ambiente coberto. Os tratamentos foram determinados a partir de diferentes manejos de controle do míldio da videira: TE - Testemunha (sem controle fitossanitário para o míldio); CA - Controle convencional (calendário); BA - Alerta fitossanitário denominado \'Regra 3-10\' (BALDACCI et al., 1947); MA25 - Alerta fitossanitário com eficiência de infecção baixa - i0 > 25% (MADDEN et al., 2000); e MA75 - Alerta fitossanitário com eficiência de infecção alta - i0 >75% (MADDEN et al., 2000). De acordo com os resultados, sob o PPT a transmissividade média da radiação solar global foi de 82,4% em 2012 e 67,3% em 2013 e sob a TP18%, da ordem de 90% nos dois anos estudados. Os ambientes sob as coberturas apresentaram temperaturas máximas do ar superiores aos valores observados a céu aberto, sendo as diferenças da ordem de 0,7 °C sob ao PPT e de 1,0 °C sob a TP18%. Sob o PPT, a duração do período de molhamento foliar foi 34% superior do que sob TP18%. Os tratamentos baseados nos sistemas de alertas fitossanitários (BA, MA25 e MA75) revelaram níveis baixos de severidade do míldio da videira sob PPT, semelhantes aos verificados no tratamento com base no calendário (CA). Os tratamentos BA, MA25 e MA75 sob TP18% indicaram um número de pulverizações similar aos realizados sob o PPT, porém foram menos eficientes em relação a CA. Houve uma significativa redução no número de pulverizações entre o tratamento CA e os tratamentos BA, MA25 e MA75, da ordem de 70%. As videiras, sob o PPT, pulverizadas com base em BA, MA25 e MA75 apresentaram características produtivas e qualitativas semelhantes às das videiras pulverizadas de acordo com CA. Conclui-se que o cultivo de videiras sob cobertura plástica de polipropileno, aliado à adoção de sistemas de alertas fitossanitários, resultou em excelentes níveis de controle do míldio da videira no Noroeste Paulista. / The Northwest region of the state of São Paulo is one of the main producers of table grapes in Brazil, however has a very favorable environmental conditions to fungal diseases during the growing season. The use of disease warning systems and plastic covers are promising alternatives for disease control, but there are not many researches about that in this region of the state. Thus, the objective of this study was to evaluate the efficacy of warning systems for managing downy mildew (Plasmopara viticola) in vineyards cultivated under plastic coverings, and, consequently, their impact on vine productivity and quality, in the northwest region of São Paulo State, Brazil. The experiment was carried out at the EMBRAPA - Tropical Viticulture Experimental Station (EVT/Embrapa Uva e Vinho), located in Jales, SP, Brazil. Three rows of 120 m of the seedless grape cultivar \'BRS Morena\', spaced with 3.0 m between plants were conducted during 2012 and 2013 growing seasons. Half of the vineyard was covered with braided polypropylene plastic film installed over a metallic arc-shaped structure (PPT) and the other half with black screen, with 18% of shading (TP18%). The experimental design was randomized blocks composed of five treatments, with six repetitions per covered environment. The treatments were defined by the different grapevine downy mildew management : (TE) Control (no sprays against downy mildew); (CA) Conventional control (calendar); (BA) Warning system \'Rule 3-10\' (BALDACCI et al., 1947); (MA25) Warning system with low-infection efficiency - i0 > 25% (MADDEN et al., 2000); and (MA75) Warning system with high infection efficiency - i0 > 75% (MADDEN et al., 2000). According to the results, under the PPT the average global solar radiation transmissivity was 82.4% in 2012 and 67.3% in 2013 and under TP18%, around 90% along the two growing seasons. The microclimate under the plastic covers showed maximum air temperatures higher than the values observed in the external environment, and the differences was around 0.7 °C under the PPT and 1.0 °C under TP18%. In the PPT, leaf wetness duration (LWD) was about 34% higher in relation to the TP18%. The treatments based on warning systems (BA, MA25 and MA75) revealed low levels of severity of grapevine downy mildew under PPT, similar to those observed in the treatment based on a calendar spray (CA). The treatments BA, MA25 and MA75 under TP18% indicated a number of sprays similar to those obtained under the PPT, but less efficient in relation to the CA. There was a significant reduction in the number of sprays between the CA and BA, MA25 and MA75 treatments, about 70%. The vines under the PPT and sprayed based on BA, MA25 and MA75 had productive and quality characteristics similar to those sprayed according to the CA. It is concluded that the association of cultivation under polypropylene plastic cover and the use of disease warning systems resulted in excellent levels of downy mildew control in vineyards in the Northwest region of São Paulo state.