No description available.

O meloeiro (Cucumis melo L.) é uma frutífera largamente cultivada no Brasil, principalmente no nordeste brasileiro, onde vem alcançando grande importância econômica, visto que grande parte da produção é voltada para a exportação. Plantas da família do meloeiro, como pepino e abóbora, são afetadas pelo oídio (Podosphaera xanthii) que causa uma das doenças foliares mais destrutivas destas espécies. Este fungo apresenta diversas raças fisiológicas e a correta identificação destas é de elevada importância para o manejo da doença, pois o melhoramento de meloeiro com o emprego de genes de resistência é o método mais eficiente para o seu controle. A identificação destas raças por meio da prática tradicional de inoculações em uma série diferenciadora de variedades de meloeiro é um método laborioso e passível de erros. Devido a isso, uma alternativa seria o uso de métodos moleculares para determinar a identidade da raça de forma rápida e econômica. O presente trabalho objetivou verificar a variabilidade de isolados de P. xanthii através da técnica de AFLP e de seqüenciamento da região ITS 5.8S do rDNA. A partir de AFLP obteve-se um dendrograma no qual não houve separação de raças, origem geográfica e nem hospedeiro. Com esta técnica verificou-se alta variabilidade entre isolados, com similaridade genética máxima de 69% e similaridade mínima de 23%. Ao contrário, os mesmos isolados apresentaram seqüências idênticas através do seqüenciamento da região ITS 5.8S do rDNA. As duas técnicas são distintas e o AFLP proporciona a obtenção de maior quantidade de fragmentos e com isso mais chances de polimorfismos. O AFLP indica que os isolados testados têm composição genética heterogênea embora isto não tenha sido evidenciado com o seqüenciamento da região ITS. / Melon (Cucumis melo L.) is a largely cultivated fruit in Brazil, especially in the northeastern region where it is achieving great economic importance since a great part of its production is destined to exportation. Plants of the melon crop family, such as cucumbers and pumpkins, are affected by powdery mildew (Podosphaera xanthii) that causes one of the most destructive foliar diseases of this species. This fungus presents various physiological races and their correct identification is of great importance to determine the control of the disease since breeding with the use of resistant genes is the most effective method for its control. Identification of these races by traditional practice of inoculation in a differential series of melon varieties is a laborious method and misleading. Due to this, an alternative would be the use of molecular methods to quickly and economically determine the race identity. The present research has the objective of verifying the variability of isolated P. xanthii through the AFLP technique and sequencing of the rDNA\'s ITS 5.8S region. From the AFLP a dendrogram was obtained where there was no separation of race, geographic region or host. With this technique, a high variability among isolated samples was verified, with 69% maximum genetic similarity and 23% minimum similarity. On the other hand, the same isolated samples presented identical sequence through the sequencing of the rDNA\'s ITS 5.8S region. The two techniques are distinct and the AFLP helps get highest amount of fragments and with this more incidence of polymorphisms. AFLP indicates that the tested isolated samples have an heterogeneous genetic composition although this was not evidenced with the sequencing of the ITS region.

No description available.

京都大学 / 新制・論文博士 / 博士(農学) / 乙第13412号 / 論農博第2895号 / 新制||農||1085(附属図書館) / 学位論文||R3||N5322(農学部図書室) / 京都大学大学院農学研究科農学専攻 / (主査)教授 冨永 達, 教授 土井 元章, 教授 那須田 周平 / 学位規則第4条第2項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Ten winter wheat (Triticum aestivum L.) lines selected from the 1982 International Winter Wheat Mildew and Rust Nurseries were studied to characterize gene number and mode of inheritance of powdery mildew resistance. Two experiments were conducted: 1) each of the lines was crossed to the susceptible cultivar Chancellor, which lacks any known’ gene _ for resistance, and seedlings of the parental lines, F₁, F₂, BC₁ (Chancellor X F₁), and F₃ populations were inoculated with isolate 127 of Blumeria graminis (DC.) E. 0. Speer f. sp. tritici Em. Marchal in the greenhouse and evaluated for powdery mildew reaction; 2) the ten lines were crossed with each other and to each of 13 host differential lines with known genes for powdery mildew resistance, and 300 to 800 F₂ seedlings from each cross were evaluated. All parents were resistant (Infection Type = 1-3), except for ST1-25, which had an intermediate (IT = 4-5) reaction type. Genetic analyses of crosses revealed that the resistance in C39 and SI5 is conferred by three dominant genes (Pm2, 4b, and 6), and resistance in A55-2, R107, and Bulk PV63-6 is governed by one partially dominant gene (Pm4b). Results from F₂, F₃, and BC₁ populations derived from crosses between ‘Armada’ and Chancellor, were inconsistent, but indicated that Armada has at least one dominant gene for resistance, which likely is Pm4b as suggested by others. The resistance gene in OK75R3645 most likely is an allele at the Pm3 locus, and it is probable that the resistance gene in GO4779 is Pm1. Single recessive genes were identified in VPM1 (Pm4b) and ST1-25 (Pm8). / Ph. D.

This study is concerned with the resistance of tobacco seedlings to a second attack of downy mildew caused by Peronospora tabacina Adam. Observations were made on the development and duration of resistance following recovery of tobacco seedlings from downy mildew. The duration of the period of resistance is variable and the observations indicate that plants in the greenhouse recovering during the summer remain resistant longer than do plants recovering from an initial attack during the winter. Samples of leaf tissue from recovered (resistant), infected and previously uninfected (susceptible) plants grown in soil in the greenhouse were analyzed for ammonia-nitrogen, nitrate-nitrogen, reducing sugars and total sugars. Similar analyses were made of samples of leaf tissue from tobacco plants grown in crocks of quartz sand and irrigated with nutrient solutions and from plants grown in crocks of sand-peat mixture to which various mixed fertilizers were added. There was no correlation between the ammonia-nitrogen content of tobacco plants, as determined in these experiments, and the response of the tobacco plant and its susceptibility to attack by P. tabacina, but there was a strong indication that such a relationship exists. Recovered plants contained a higher ratio of total sugar to nitrate-nitrogen than did comparable plants which had never been infected. Furthermore the ratio of total sugar to nitrate-nitrogen was lower in plants infected for a second time than it was in plants from the same lot shortly after they had recovered from the initial attack. It seems probable however that the changes in the nitrate-nitrogen and sugar content in recovered tobacco leaves are an indication of recovery and are not responsible for the resistance possessed by such plants. A series of experiments were performed to test the immediate effect of several nutrient salts and of sugar upon the sporulation of the fungus on the leaves of diseased tobacco plants. There was a very definite response in sporulation to some of these treatments, especially to potassium nitrate, calcium nitrate, and sucrose, but it is not known whether the response of sporulation of the fungus to such treatment is related to the type of resistance possessed by recovered plants. Sodium chloride and calcium chloride were dissolved in water and applied to the soil in which young tobacco plants were growing in the greenhouse. Although increased chloride in one of the sand cultural experiments seemed to protect the plants somewhat against downy mildew, application of solutions of these salts to the soil at the rates used (100 lbs. And 138 lbs. Per acre respectively) produced no noticeable effect on the severity of the disease or subsequent recovery of the plants. The leaves of recovered (resistant) tobacco plants are usually reduced in number, the lower ones having been killed by the initial attack of the fungus. Removal of the leaves from healthy tobacco plants in imitation of this condition did not produce resistance to an attack by P. tabacina. The artificial production of necrotic areas on healthy tobacco leaves in simulation of those usually following sporulation of P. tabacina on diseased leaves had no effect either on the susceptibility of the plants to infection or upon sporulation of the fungus on leaves already infected at the time the necrotic areas were induced. Results of attempts to confer immunity from downy mildew upon an entire plant by infection of a single leaf of the plant were inconclusive, since under the conditions of the experiment it was impossible to be sure that the single leaf was infected. However, the indication is that infection of a single leaf will not render the entire plant immune from a second attack by the fungus. / Master of Science

In the United States, hops (Humulus lupulus L.) are grown mainly in the Pacific Northwest (PNW). For this reason, most cultural information is based on the growing conditions of the PNW. Growing conditions in Virginia differ drastically and present unique disease and production challenges. Three studies were conducted with the intent of increasing hop cultivation knowledge for Virginia growers. For the first study, 13 cultivars of hops grown at the Virginia Tech hop yard were compared for growth, yield, and quality. Mean cone fresh weight per plant ranged from 12.00 g for Mt. Hood to 1002.87 g for Crystal in 2016 and from 97.98 g for Mt. Hood to 900.33 g for Cascade in 2017. In 2016, only Alpharoma, Cascade, Mt. Rainier, and Southern Cross had alpha acid levels, an indicator of cone quality, within the accepted range. In 2017, Alpharoma, Centennial, Mt. Rainier, and Nugget had alpha acid levels within the expected range. Three cultivars (Cascade, Crystal, and Ultra) were above the expected alpha acid range, which indicates more bittering potential for beer brewers. In a nutrient deficiency study, hop plants were grown in hydroponic solutions, and deficiencies were induced for nitrogen (N), phosphorus (P), and potassium (K). After visual deficiency symptoms had been induced, leaf tissue samples were taken and analyzed for nutrient content. Images were taken at each deficiency stage. For N, incipient deficiency symptoms were observed at a mean of 3.18% dry weight in leaf tissue samples. Visual symptoms included a chlorotic appearance, undersized leaves, and red petioles. Incipient symptoms for P were observed at a mean of 0.307% dry weight in leaf tissue samples. Necrotic spots, leaf cupping, and undersized leaves were apparent with this deficiency. Incipient symptoms for K were observed at a mean of 1.21% dry weight in leaf tissue samples. Symptoms included rounded leaf tips, blue veins, and marginal scorch. In the third study, a whole leaf powdery mildew (Podosphaera macularis) assay was developed and tested using five hop cultivars, Alpharoma, Cascade, Comet, Sorachi Ace, and Tahoma. Leaves were inoculated with powdery mildew (PM) using a settling tower. This method was used to rapidly assess the resistance of cultivars. Leaves were successfully inoculated and PM colonies were allowed to grow for two weeks. Images of the PM colony development on inoculated leaves were compared using ImageJ to determine percentage of coverage. Tahoma was the only cultivar found to produce a significantly different mean percent coverage (19.5%) compared with the resistant cultivars Cascade and Comet (<1%). / Master of Science in Life Sciences / Hops (Humulus lupulus) were first grown in the United States in Massachusetts in the early 1600s. Production of this crop eventually spread throughout the Northeastern US. By the mid-1800s, commercial production spread to southern states such as Virginia. Infestation of pests, e.g., hop aphid and diseases such as downy mildew in eastern states, and prohibition on the production of alcohol caused a significant shift in hop production, which favored the Pacific Northwest (PNW). As a result, in Virginia specific knowledge of hop cultivation is now lacking. Three studies were conducted to increase our knowledge of hop cultivation for the region. For the first study, 13 cultivars grown at the Virginia Tech hop yard were compared for growth, yield, and quality. The cultivars Cascade and Alpharoma had alpha acid levels equal to or higher than expected for the 2016 and 2017 seasons. The alpha acid levels are an indicator of the hop quality from the Cascade and Alpharoma cultivars. Cascade was also a top producer of hop cones with a mean fresh weight of 989.67 g and 900.33 g in 2016 and 2017, respectively. In a nutrient deficiency study, plants were grown in Hoagland hydroponic solutions, which contain all essential nutrients needed for plant growth. This was done to provide photographic documentation to assist local growers with deficiency identification. Deficiencies were induced for nitrogen (N), phosphorus (P), and potassium (K) by removing each of the specific nutrient of interest from each treatment solution. After deficiency symptoms were induced, leaf tissue samples were taken and analyzed for nutrient content. Nutrient deficiencies were documented using photographs. For N, visual symptoms included a chlorotic appearance, undersized leaves, and red petioles. Incipient symptoms for P included necrotic spots, leaf cupping, and undersized leaves. For K, leaf cupping, blue green veins, marginal scorch, and rounded leaf tips. Disease resistance of cultivars is important information for growers and can heavily influence hop yard planning. As such, in another study, an assay for powdery mildew (Podosphaera macularis) was developed that allows for rapid low cost testing of hop cultivars. The assay was tested on the following cultivars: Alpharoma, Cascade, Comet, Sorachi Ace, and Tahoma. Leaves were inoculated with powdery mildew (PM) using a settling tower. Powdery mildew colonies were allowed to grow for two weeks and were then analyzed using ImageJ software to determine percent coverage. At the end of the experiment, Tahoma had significantly greater PM coverage compared to the other cultivars, indicating that Tahoma is less resistant to the specific PM strain. / Hops (Humulus lupulus) were first grown in the United States in Massachusetts in the early 1600s. Production of this crop eventually spread throughout the Northeastern US. By the mid-1800s, commercial production spread to southern states such as Virginia. Infestation of pests, e.g., hop aphid and diseases such as downy mildew in eastern states, and prohibition on the production of alcohol caused a significant shift in hop production, which favored the Pacific Northwest (PNW). As a result, in Virginia specific knowledge of hop cultivation is now lacking. Three studies were conducted to increase our knowledge of hop cultivation for the region. For the first study, 13 cultivars grown at the Virginia Tech hop yard were compared for growth, yield, and quality. The cultivars Cascade and Alpharoma had alpha acid levels equal to or higher than expected for the 2016 and 2017 seasons. The alpha acid levels are an indicator of the hop quality from the Cascade and Alpharoma cultivars. Cascade was also a top producer of hop cones with a mean fresh weight of 989.67 g and 900.33 g in 2016 and 2017, respectively. In a nutrient deficiency study, plants were grown in Hoagland hydroponic solutions, which contain all essential nutrients needed for plant growth. This was done to provide photographic documentation to assist local growers with deficiency identification. Deficiencies were induced for nitrogen (N), phosphorus (P), and potassium (K) by removing each of the specific nutrient of interest from each treatment solution. After deficiency symptoms were induced, leaf tissue samples were taken and analyzed for nutrient content. Nutrient deficiencies were documented using photographs. For N, visual symptoms included a chlorotic appearance, undersized leaves, and red petioles. Incipient symptoms for P included necrotic spots, leaf cupping, and undersized leaves. For K, leaf cupping, blue green veins, marginal scorch, and rounded leaf tips. Disease resistance of cultivars is important information for growers and can heavily influence hop yard planning. As such, in another study, an assay for powdery mildew (Podosphaera macularis) was developed that allows for rapid low cost testing of hop cultivars. The assay was tested on the following cultivars: Alpharoma, Cascade, Comet, Sorachi Ace, and Tahoma. Leaves were inoculated with powdery mildew (PM) using a settling tower. Powdery mildew colonies were allowed to grow for two weeks and were then analyzed using ImageJ software to determine percent coverage. At the end of the experiment, Tahoma had significantly greater PM coverage compared to the other cultivars, indicating that Tahoma is less resistant to the specific PM strain.

L'utilisation des champignons mycorhiziens à arbuscules (CMA) pourrait constituer une alternative potentielle aux traitements fongicides conventionnels pour lutter contre les maladies cryptogamiques des plantes. Notre travail a consisté à étudier l'éventuel effet protecteur de la mycorhization arbusculaire chez le blé tendre (triticum aestivum L.) contre Blumeria graminis f. sp. tritici (Bgt), un champignon biotrophe responsable de l'oïdium, une maladie affectant les parties aériennes de la plante. L'inoculation mycorhizienne du blé avec le CMA Funneliformis mosseae (FM), en conditions controlées et optimisées, nous a permis d'obtenir parallèlement à un taux de mycorhization de 38% des plants de blé, une amélioration significative de la biomasse et un taux de protection contre Bgt estimé à 78%. Ces résultats suggèrent l'induction d'une résistance systémique des réactions de défense du blé par mycorhization (Mycorrhiza-Induced Resistance, MIR). Cette protection serait liée à une accumulation de composés phénoliques et de preoxyde d'hydrogène dans les cellules épidermiques des feuilles de blé mycorhizé, au niveau du site de pénétration de Bgt. Une surexpression des gènes POX, PAL, CH11 et NPR1 codant pour des marqueurs de défense a également été mise en évidence dans les feuilles en absence d'infection par Bgt. Enfin, nos travaux ont également souligné l'intégration de divers paramètres pour optimiser l'utilisation des CMA comme agents de biocontrôle chez le blé. La meilleure protection contre l'oïdium a été obtenue aec un apport en phosphore réduit de 5 fois par rapport à celui préconisé au champ et un inoculum mycorhizien à base de Fm, que ce soit chez un cultivar modérément sensible ou un cultivar plus résistant. / The use of arbuscular mycorrhizal fungi (AMF) could be an innovative alternative to chemicals against fungal plant diseases. Our work aimed at studying the possible protective effect of arbuscular mycorrhization in the bread wheat (Triticum aestivum L.) against Blumeria graminis f. sp. tritici (Bgt), a biotrophic fungi responsible of wheat powdery mildew, a disease affecting the aerial plant organs. Wheat mycorrhizal inoculation by Funneliformis mosseae (Fm), under controlled and optimized conditions, allowed us to obtain concomitantly a micorrhizal rate of 38%, a significant increase of plant biomass and a protection level against Bgt estimated at 78%. These results suggest the induction of systemic wheat defense reactions resulting from mycorrhization (Mycorrhiza-Induced Resistance - MIR). This protection is linked to an accumulation of phenolic compounds and hydrogen peroxide at the Bgt penetration sites in epidermal leaf cells of mycorrhized wheat plants. Up-regulations of POX, PAL, NPR1 and CH11 genes encoding for defense markers were also pointed out in leaves of mycorrhizal wheat in the absence of Bgt infection. Moreover, our study highlighted the importance of taking into account various parameters to optimize the use of AMF as biocontrol agents. The highest protection against powdery mildew was obtained with a 5-fold reduced phosphorus input compared to that recommended in the field and with the mycorrhizal inoculum Fm, in both a moderately susceptible or a more resistant cultuva

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.

At present, the global population is increasing, while soil and fresh water resources for crop production are declining. It is important to adopt sustainable practices to optimise the use of limited natural resources without compromising the environment, and to enhance continuous production in the long term. The rapid growth of UK strawberry industry has been achieved through the precision use of varieties, nutrients and polythene tunnels. This intensive production has caused significant environmental impacts especially Greenhouse Gas (GHG) emissions from the production. Strawberry powdery mildew (Podosphaera aphanis) is a major fungal disease affecting strawberry production worldwide particularly in polythene tunnels. The disease can result in yield losses of up to 70% of the crop. A ruleQbased system was used in the field trials to predict high risk days of P. aphanis development, taking into account the optimal environmental conditions conducive to conidial germination and disease development. The results (Chapter 3) showed that the use of this prediction system achieved satisfactory control of P. aphanis in commercial strawberry production, with reduced fungicide applications compared with commercial spray programme. The results were consistent in two consecutive years and on different varieties. In addition, it was suggested that the use of the prediction system may also lead to lower GHG emissions associated with fewer fungicide applications, thereby benefit strawberry growers both environmentally and economically. Results from 2014 & 2015 silicon fertigation trials showed that the use of a silicon nutrient via the fertigation system reduced the strawberry susceptibility to P. aphanis and twoQspotted spider mites (Tetranychus urticae Koch) in two consecutive years on different varieties (Chapter 4). In both years, crops received the silicon nutrient only without fungicides had both lower rate of epidemic (r) and lower value of Area Under the Disease Progress Curve (AUDPC) (r = 0.0036, AUDPC = 475 in 2014; r = 0.001, AUDPC = 267 in 2015) compared with the untreated control (r = 0.0042, AUDPC = 662 in 2014; r = 0.0011, AUDPC = 281 in 2015). Silicon also delayed the epidemic buildQup in the silicon nutrient only treatment for approximately two weeks compared with the untreated control. Crops from the silicon nutrient plus fungicides treatment had lower susceptibility (r = 0.0012 in 2014; r = 0.0004 in 2015) than those from the fungicides only treatment (r = 0.0017 in 2014; r = 0.0005 in 2015) suggesting that the silicon nutrient may also enhance fungicides performance in reducing the epidemic buildQup when used together. Moreover, the presence of T. urticae on strawberry leaves was significantly lower (P < 0.001) in plants treated with the silicon nutrient than those without. In addition, initial results suggested that silicon may play a positive role in raising °Brix of strawberry leaf petiole, improving pollen viability, and influencing the length of flower receptacle and stamens. Maltmas Farm has a wide range of semiQnatural habitats that provide food and nesting resources for wild pollinators. Hoverflies, bumblebees and solitary bees were found to be the main wild pollinators that pollinate commercial strawberries at Maltmas Farm (Chapter 5). The number of pollinators in tunnels or open fields significantly correlated with the abundance of strawberry flowers (P < 0.05). Pollinator presence also differed between groups throughout the day and over the seasons. Hoverflies appeared early in the day and were abundant in summer months; bumblebees and solitary bees were present most of the day and throughout the season, whereas honeybees were only active in sunny days. Temperatures, relative humidity and cloud coverage also affected pollinator presence. In addition, pollinator activity was not significantly (P > 0.05) affected by the application of the silicon nutrient via the fertigation system. The integrated use of the prediction system (to reduce fungicide applications and subsequent GHG emissions), the silicon nutrient (to reduce crop susceptibility to P. aphanis and T. urticae), and sustainable farmland management (to encourage the presence of wild pollinators) could help strawberry growers to achieve a more sustainable production.