Sclerotinia homoeocarpa F.T. Bennett, the causal agent associated with dollar spot, is a common and economically challenging turfgrass disease in North America. Acceptable turfgrass quality requires the optimization of cultural practices and the judicious use of fungicides. Sclerotinia homoeocarpa causes significant damage to turfgrass swards from May to October annually, therefore, requiring multiple fungicide applications to maintain satisfactory turfgrass quality throughout the growing season. Sterol demethylation inhibitor (DMI) fungicides are among the most widely used in the United States and frequent use has led to the development of fungicide resistance to the DMI fungicide class. The precise mechanism of DMI fungicide resistance in S. homoeocarpa is not completely understood, however over expression of the CPY51A gene or efflux transporter genes have been reported as the molecular mechanism for other fungal systems. Fungicide resistance to the DMI class exhibits a gradual population shift towards insensitivity that can be monitored using In vitro fungicide sensitivity assays. In vitro fungicide sensitivity assays have been used to detect fungicide sensitivity differences in different S. homoeocarpa isolates and currently serve as the most accurate methodology to detect DMI insensitivity in S. homoeocarpa. Documentation of DMI field efficacy on native S. homoeocarpa populations with differing in vitro sensitivities has not been examined thoroughly and would provide critical information regarding the association between in vitro fungicide sensitivity and DMI field efficacy. The objectives of this research are (i) to determine the association between in vitro propiconazole sensitivity and reduced field efficacy for five native S. homoeocarpa populations and (ii) to develop a qualitative in vitro sensitivity assay for detection of S. homoeocarpa isolates responsible for practical field resistance.

Cette étude vise à mettre en évidence, à l'aide de méthodes d'inoculation dirigée, des différences de résistance à l'infection par le Sclerotinia sclerotiorum entre des lignées de soja disponibles au Québec. Pour ce faire, les objectifs suivants ont été fixés: 1) préciser la résistance ou la sensibilité à la sclérotiniose chez 30 lignées ou cultivars de soja adaptés au Québec; 2) examiner la contribution de la fleur (pétales ou axe floral) et de la tige (intacte ou blessée) à la réaction des lignées ou cultivars; 3) examiner l'efficacité ou la complémentarité de techniques d'inoculation caulinaire ou florale, avec du mycélium ou des ascospores, au champ ou en serre, comme méthodes d'étude de la réaction de lignées de soja au S. sclerotiorum. Les expériences ont été menées à l'Université Laval. En 1998, le mycélium a été utilisé dans trois techniques d'inoculation: inoculation des tiges blessées en serre et au champ, des tiges intactes au champ et des fleurs au champ. Les ascospores collectées de sclérotes produits et conditionnés au laboratoire ont été utilisées à l'hiver 1999 dans une technique d'inoculation des fleurs en serre. L'inoculation de tiges blessées, en serre, a permis de distinguer 12 lignées significativement moins sensibles à la colonisation de la tige blessée que le cultivar Nattosan, le plus sensible. L'inoculation d'ascospores en suspension, en serre, met en évidence deux cultivars significativement moins sensibles à l'invasion des inflorescences (Maple Donovan et Frisquet). Cette recherche montre des différences significatives de résistance à la sclérotiniose chez des lignées de soja du Québec. On précise également, grâce aux inoculations dirigées, la nature de la résistance ou de la sensibilité de quelques lignées: florale (ex: Maple Donovan et Frisquet) ou caulinaire (ex: Nattosan). Ces inoculations dirigées ont présenté une forte variabilité mais elles sont suffisamment discriminantes et complémentaires pour opérer une première sélection de parents partiellement résistants utiles à un programme d'amélioration génétique de la résistance des lignées de soja du Québec au S. sclerotiorum.

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Made available in DSpace on 2014-07-29T14:42:44Z (GMT). No. of bitstreams: 1 claudia adriana.pdf: 420834 bytes, checksum: 841b3fc76b59f3d3b0619cb8de80fc1a (MD5) Previous issue date: 2009-08-10 / The objective of this study was to evaluate the association of cultural practices established with soil mulch with straw of Brachiaria ruziziensis, and biological control with sprayings of 2 x 109 viable spores / mL of Trichoderma harzianum strain 1306, to decrease the inoculum density of Sclerotinia sclerotiorum, causal agent of white mold on soybeans, and the incidence of white mold. The experiment was carried out in Jataí, GO, at the Brazilian Center-West Region, in a commercial soybean field naturally infested with the pathogen, in 2006/2007 and 2007/2008 seasons. Implemented in March 2006 on plots of 630 m² in a completely randomized design with three repetitions, the presence and absence of B. ruziziensis and applications of T. harzianum in dosages 0, 0.5, 1.0 and 1.5 L.ha-1 in one or two sprayings (March and October). The soil was sampled for quantification of sclerotia in March to assess the initial inoculum, in June, and September to evaluate the viability of sclerotia on PDA medium. Field and laboratory assessments were done for viability and parasitism of S. sclerotiorum sclerotia by Trichoderma spp. and other soil microorganisms, the number of apothecia, white mold incidence and yield components. In the presence of B. ruziziensis the viability of sclerotia was lower and at 0.5 and 1.0 L.ha-1 T. harzianum strain 1306 parasitism of sclerotia was higher than the control without T. harzianum strain 1306 . In the presence of straw in Brachiaria decreased the number of apotécios. The white mold incidence of residual effects was lower in the presence of straw in doses 0.5 and 1.0 L.ha-1 T. harzianum strain 1306 . The grain yield of soybean was higher dosage of 0.5 L ha-1 T. harzianum strain 1306 . / O objetivo desta pesquisa foi avaliar o efeito integrado dos métodos de controle cultural, pela semeadura de Brachiaria ruziziensis para formação de palha no Sistema Plantio Direto, e controle biológico com aplicação de Trichoderma harzianum 1306 , produto comercial Trichodermil®, na redução da densidade de inóculo e na incidência de Mofo branco. Realizado no município de Jataí, GO, em área comercial de soja naturalmente infestada por Sclerotinia sclerotiorum nas safras 2006/2007 e 2007/2008. Implantado em março de 2006 em faixas com parcelas de 630 m², na presença e ausência de B. ruziziensis e aplicações de T. harzianum 1306 nas dosagens 0, 0.5, 1.0 e 1.5 L.ha-1 em uma ou duas épocas (março e outubro). O solo foi amostrado para quantificação de escleródios, em março para avaliação inicial do inóculo, em junho, e em setembro para avaliação de viabilidade de escleródios em meio BDA. Foram avaliados o número de escleródios, a viabilidade e o parasitismo de escleródios por Trichoderma spp e por outros microorganismos, o número de apotécios, a incidência da doença e os componentes de produção. A palhada de braquiária reduziu a viabilidade de escleródios e o número de apotécios, e ainda, nas doses de 0,5 e 1,0 L.ha-1 de T. harzianum 1306 o parasitismo de escleródios foi superior. A incidência de Mofo Branco foi menor no efeito residual da presença de palhada de B. ruziziensis nas doses 0,5 e 1,0 L.ha-1 de T. harzianum 1306 . A produtividade de grãos de soja foi maior na dosagem de 0,5 L ha-1 de T. harzianum 1306 .

The biological control agent Pseudomonas chlororaphis PA23 is effective at protecting Brassica napus (canola) from the necrotrophic fungus Sclerotinia sclerotiorum via direct antagonism. Despite the growing importance of biocontrol bacteria in protecting crop plants from fungal pathogens, little is known about how the host plant responds to bacterial priming on the leaf surface and certainly nothing about global changes in gene activity in the presence and absence of S. sclerotiorum. PA23 priming of mature canola plants reduced the number of lesion-forming petals by 90%. Global RNA sequencing of canola tissue at the host-pathogen interface showed a 16-fold reduction in the number of genes uniquely upregulated in response to S. sclerotiorum when pretreated with PA23. Upstream defense-related gene patterns suggest MAMP-triggered immunity via surface receptors detecting PA23 flagellin and peptidoglycans. Although systemic acquired resistance (SAR) was induced in all treatment groups, a response centered around a glycerol-3-phosphate (G3P)-mediated pathway was exclusively observed in canola plants treated with PA23 alone. Activation of these defense mechanisms by PA23 involved production of reactive oxygen species as well as pronounced thylakoid membrane structures and plastoglobule formation in leaf chloroplasts. PA23 therefore primes defense responses in the plant through the induction of unique local and systemic regulatory networks. / October 2016

Plant programmed cell death (PCD) is an essential process in plant-pathogen interactions. Importantly, PCD can have contrasting effects on the outcome depending on context. For example, plant PCD in plant-biotroph interactions is clearly beneficial to plants, whereas it could be detrimental to plants in plant-necrotroph interactions. Sclerotinia sclerotiorum is an agriculturally and economically important necrotrophic pathogen. Previous studies have shown that S. sclerotiorum secretes oxalic acid (OA) to enhance Sclerotinia virulence by various mechanisms including induction of PCD in plants. A recent study has also shown that reactive oxygen species (ROS) generation correlates with induction of PCD during disease development. These studies focus on links between ROS, oxalate, and PCD, and how they impact S. sclerotiorum disease development. I examined the involvement of ROS in pathogenic development of S. sclerotiorum. I identified and functionally characterized two predicted S. sclerotiorum NADPH oxidases (Nox1 and Nox2) by RNAi. Both nox genes appear to have roles in sclerotial development, while only Nox1-silenced mutants showed reduced virulence. Interestingly, the reduced virulence of the Nox1-silenced mutant correlated with decreased production of OA in the mutant. This observation suggests that regulation of ROS by S. sclerotiorum Nox1 may be linked to OA. The next study details the phenotype of plants inoculated with an S. sclerotiorum oxalate deficient mutant (A2), which showed restricted growth at the infected site. This response resembles the hypersensitive response (HR), and is associated with plant resistance responses including cell wall strengthening, plant oxidative burst, and induction of defensin genes. Conversely, leaves infected with wild type showed unrestricted spreading of cell death and were not associated with these resistant responses. Furthermore, previous work had shown that a Caenorhabditis elegans anti-apoptotic gene (ced-9) conferred resistance to wild type S. sclerotiorum, while this gene had negligible effects on the phenotype of plant leaves inoculated with A2 mutants. These findings suggest that HR-like cell death by A2 and PCD by wild type S. sclerotiorum may be regulated by different pathways. As a whole, these results reveal the importance of ROS, oxalate, and PCD in Sclerotinia disease development as well as the significance of interplay between them. These studies contribute to the understanding of the underlying mechanisms of Sclerotinia disease.

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Brassica napus ist eine der wichtigsten Pflanzen in der Landwirtschaft. Pathogene verursachen jährlich große Ertragsverluste. Im Rahmen dieser Arbeit wurden die Interaktionen zwischen der Nutzpflanze Brassica napus bzw. der Modellpflanze Arabidopsis thaliana und dem phytopathogenen Pilz Verticillium longisporum untersucht. Verticillium longisporum ist ein auf Brassica spezialisierter Pilz, der die sogenannte Verticillium-Welke verursacht. Er dringt in die Wurzeln der Pflanze ein und steigt durch die Leitgefäße in den Spross auf. Die Pflanze zeigt in Folge der Infektion vor allem einen stark verkürzten Spross, Chlorosen und eine Notreife. Für die Experimente wurde Brassica napus und Arabidopsis thaliana mit Verticillium longisporum infiziert. Die Emissionen von Volatilen (volatile organic compounds (VOC)) der Pflanzen wurden sowohl ober- als auch unterirdisch bezüglich sich ändernder VOCs analysiert. Bei der Probennahme wurde darauf geachtet, die Pflanzen nicht zu verletzen und durch Messungen bedingten Stress zu minimieren. Auch während der Messungen der Wurzel-emissionen wurden die Pflanzen nicht aus ihrem Substrat entfernt, sondern innerhalb dessen gemessen. Während sich das Duftspektrum der Wurzeln von Brassica napus nicht änderte, emittierten die Sprosse infizierter Brassica Pflanzen signifikant mehr Dimethyldisulfid, ß-Ionon und ß-Cyclocitral. Bei der Infektion von Arabidopsis thaliana mit Verticillium longisporum hingegen spielten diese VOCs keine Rolle. Im zweiten Teil der Arbeit wurde die Vermutung überprüft, dass sich im Infektionsverlauf ändernde VOCs als Abwehrmoleküle dienen könnten. Hierfür wurde ein Bioassay entwickelt und zwei weitere pathogene Pilze, Gaeumannomyces graminis var. triciti und Sclerotinia sclerotiorum, unterschiedlicher Spezialisierung einbezogen. Wir setzten sie Dimethyldisulfid und ß-Ionon in unterschiedlichen Konzentrationen aus. Sowohl Dimethyldisulfid als auch ß-Ionon zeigten nur eine geringe fungizide Wirkung, gemessen am Myzelwachstum, gegen den Brassica-Spezialisten Verticillium longisporum. Gaeumannomyces graminis var. triciti wurde bereits bei geringeren Konzentrationen gehemmt. Sclerotinia sclerotiorum reagierte nicht oder sogar positiv auf die VOCs. Die (Mikro-) Sklerotienbildung wurde nur bei Verticillium longisporum beeinflusst. ß-Ionon unterstützte diese Entwicklung signifikant.