Reports concerning the life history of Plasmodiophora brassicae Wor. are highly conflicting. The contradictions in the literature on the subject and the difficulties encountered in the initial phase of the present investigation brought about an inquiry into the techniques employed by previous workers. Repetition of these techniques indicated that many previous experiments had not taken into consideration the problems inherent in an in vitro study of Plasmodiophora brassicae. Conclusions drawn from such experiments are thus of doubtful value. Methods described as successful by earlier investigators were employed in an attempt to stimulate germination. The germination process was not observed, and contaminants introduced with the resting spore material made it impossible to designate any one motile organism as P. brassicae. Attempts were made to obtain root hair infection. Pour methods described by previous investigators proved unsuccessful. A limited degree of infection was obtained by placing young cabbage and cauliflower seedlings in a buffered nutrient solution containing washed P. brassicae resting spores. In spite of the fact that artifacts were readily produced, early infection stages were observed and photographed and the existence of a zoosporangial stage in the root hair was confirmed. The infection rate was too low to permit intensive observations of the development of the parasite within the root hair. A proteolytic enzyme preparation was somewhat successful in increasing the infection rate. This was taken as an indication that a combination of enzymes could provide the necessary germination stimulus. Decay of the host tissues is apparently necessary to the maturation of the resting spores. Present experiments indicated that it is not feasible to separate the contaminants from these spores. As it was not possible to draw conclusions from contaminated cultures, it is concluded that the only approach of value is to provide an artificial germination stimulus to spores from clean clubs, so that the development of the parasite may take place under sterile conditions. / Land and Food Systems, Faculty of / Graduate

No description available.

Clubroot disease, caused by Plasmodiophora brassicae, poses a threat to the Canadian canola industry, and breeding of resistant cultivars is urgent. The objective of this study was to identify Brassica germplasm possessing resistance to Canadian P. brassicae pathotypes based on greenhouse screening. Germplasm resistant to local pathotype(s) is the prime requirement for breeding clubroot resistant cultivars. Race-specific to broad-spectrum resistance was identified in the diploid species B. rapa (AA) and B. oleracea (CC), and in the amphidiploid B. napus (AACC). The diploid B. nigra (BB) also showed race-specific to broad-spectrum resistance; however, the two amphidiploids with B. nigra as one of the parental species viz., B. juncea (AABB) and B. carinata (BBCC) were completely susceptible. The occurrence of resistance in the diploid and amphidiploid Brassica species is discussed in the light of their evolution, and a differential set for identification of Canadian clubroot pathotypes is proposed. / Plant Science

Thesis (Ph. D.)--University of Wisconsin--Madison, 1968. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1942. / Typescript. Includes abstract and vita. eContent provider-neutral record in process. Description based on print version record. Bibliography: leaves 47-49.

The aim of this project was to develop treatments that would be able to reduce the survival of clubroot spores in field soil and protect the roots of young transplants against infection. The project focused on using combinations of treatments which integrate novel and existing controls with emphasis placed on the sustainable use of waste materials, plant materials and bioactives. A wide range of treatments were screened individually and in combination under glasshouse and field conditions, e.g. fungicides, nutritional amendments, companion planting, plant saponins and biocontrol agents. Many of these treatments were able to reduce clubroot severity to varying degrees. Glasshouse treatments were more successful at controlling disease than those applied in the field. The most effective treatments - when applied correctly - contained calcium, e.g. lime as calcium oxide or LimeX (a by-product of the sugarbeet processing industry), and crushed scallop and whelk shells (a by-product of the fishing industry). Whilst the effects of calcium and pH on clubroot are not new, growers need to think more in terms of dose of calcium applied in the field rather than just pH, and also, the time of addition of lime to soil before planting needs serious consideration as it may be optimal to apply lime less than two weeks before transplanting. Experimental results have shown that soil microflora plays a major role in the development of clubroot disease and that the membrane potential of growing roots may be one of the most important factors in preventing P. brassicae from entering plant roots and causing disease due to the effect that calcium and pH have on clubroot control. The experiments have also shown that there are interactions between treatments such as fungicides, limes, soil nutritional level and soil microflora at controlling disease and some treatments may reduce the effectiveness of other treatments at controlling disease. Another factor that is known to affect the effectiveness of treatments at controlling clubroot is the initial spore load in the soil. Therefore, an additional aim of the project was to develop a rapid, quantitative PCR based diagnostic test that could measure the level of clubroot spores directly from soil. Plasmodiophora brassicae DNA was successfully extracted and amplified from artificially inoculated soils and from naturally infested field soils using real-time PCR with selected sets of primers and probes. Many different types of soil DNA extraction methods were tested and standard curves relating to different levels of spore inoculum were created. This project has generated useful information as to why there are contradictory results in clubroot research about the effect of various treatments at controlling clubroot. This information may also be the basis of practical advice to brassica growers on best practices to use to achieve optimal clubroot control in the field. Options in relation to new sustainable control treatments are discussed in the light of the results from both glasshouse and field experiments. These involve planting brassicas on raised beds and applying treatments strategically around the root zone. LimeX 70 or powdered calcium oxide were demonstrated to be the most optimal lime treatments for control, and a split application of Perlka (granular calcium cyanamide) may prove to be a consistently effective method for controlling clubroot.

Master of Science

Des études récentes ont montré que la variabilité de l’épigénome des plantes est un acteur important dans la réponse des plantes aux stress abiotiques et biotiques. La hernie, causée par le protiste Plasmodiophora brassicae, est une maladie racinaire majeure des Brassicaceae cultivées dont la résistance quantitative est considérée comme résultant principalement de la ségrégation de multiples allèles. L'objectif de ma thèse est d'établir s'il existe, chez Arabidopsis thaliana, une variabilité épigénétique héritable à l'origine de variations de la réponse à l’infection par la hernie. Pour répondre à cet objectif, une approche non ciblée d’épigénétique quantitative a été réalisée en utilisant la population épiRIL ddm1-2 x Col-0. Dix-sept QTL sous contrôle épigénétique (QTLépi), regroupés en 6 régions génomiques, ont ainsi été détectés, 5 d’entre eux étant sous la dépendance de la température.Finalement, deux régions identifiées comme impliquées dans la réponse à la hernie ont été caractérisées plus finement. La région du gène majeur de résistance à la hernie RPB1, qui colocalise avec 3 QTLépi, présente une variation génomique prépondérante dans les écotypes d’Arabidopsis potentiellement due à des mouvements d’éléments transposables. Le QTL Pb-At5.2 est sous le contrôle d’une épimutation régulant l’état de méthylation et l’expression de deux gènes NLR. Les résultats obtenus montrent que la résistance quantitative à la hernie est associée à des variations de la méthylation de l’ADN stables et héritables suggérant un modèle complexe de régulation de la résistance où la combinai / Recent studies have shown that plant epigenome variability is an important factor in plant response to abiotic and biotic stress. Clubroot caused by the protist Plasmodiophora brassicae is a major disease of Brassicaceae whose quantitative resistance is supposed to result from many allele segregation. The aim of my work is to understand if, in Arabidopsis thaliana, an inherited epigenetic variability can lead to variations in clubroot resistance. For that, an untargeted approach of quantitative epigenetics was carried out using the epiRIL population ddm1-2 x Col-0. Seventeen QTL under epigenetic control (QTLepi), clustered in 6 genomic regions, were detected, 5 of them being temperature-dependant.Finally, two regions previously identified as involved in clubroot response were finely studied. The region of the major clubroot resistance gene RPB1, which colocalizes with three QTLepi, shows major genomic variations in Arabidopsis ecotypes potentially due to movements of transposable elements. The QTL Pb-At5.2 is depending on one epimutation controlling the methylation state and the expression of two NLR genes. The results obtained demonstrate that the clubroot quantitative resistance is associated with inherited stable DNA methylation variations suggesting a complex model of resistance regulation where favourable alleles and epialleles association is necessary to obtain an optimal resistance.

La hernie est une maladie racinaire affectant les Brassica, causée par Plasmodiophora brassicae. Le déploiement de variétés présentant des facteurs génétiques de résistances est le moyen de lutte principal contre cette maladie. Toutefois, l’efficacité de ces variétés résistantes dans les nouveaux systèmes d’agriculture durable, à bas intrant azoté, est encore à évaluer. Dans ce contexte, les questions de recherche posées dans cette thèse sont les suivantes : i) La modulation de la résistance à la hernie par la réduction de l’apport azoté est-elle génotype/isolat dépendante ? ii) Y-a-t-il un effet de la réduction des apports azotés sur l’architecture génétique de la résistance à la hernie chez le colza ? iii) Quels sont les réponses transcriptomiques et métabolomiques impliquées dans la réduction du développement des symptômes en condition de carence azotée ? Le criblage de la réponse de nombreux génotypes de colza vis-à-vis de plusieurs isolats de P. brassicae a été réalisé dans deux conditions de fertilisation azotée (8 mM et 1 mM de nitrate). Basé sur ces expériences, le contrôle génétique du développement des symptômes et de la production des spores de repos lors de l’infection par deux isolats (eH et K92-16) a été étudié par génétique de liaison chez une population biparentale et par génétique d’association chez un panel de colza d’hiver. Enfin, les réponses moléculaires impliquées dans la réduction du développement des symptômes observée chez le génotype ‘Yudal’ en condition 1mM ont été étudiées par des méthodes de RNA-seq et de métabolomique. L’impact de la fertilisation azotée sur la quantité de symptômes et la production de spores de repos est variable selon la combinaison génotype/isolat. Les modulations observées entraînent une augmentation de la résistance en condition 1mM. La fertilisation azotée module également l’effet (R2) de certains des Quantitative Trait Loci (QTL) impliqués dans la résistance. Dans le cas de l’isolat eH, elle affecte principalement l’effet des QTL C09 et C02. L’augmentation de résistance observée chez le génotype ‘Yudal’ en condition 1mM, vis-à-vis de l’isolat eH, semble liée à la surexpression de gènes impliqués dans l’assimilation des nitrates (famille NRT2) et dans le transport de l’auxine. / Clubroot is a Brassica root disease caused by Plasmodiophora brassicae. The use of varieties including genetic resistance factors is the most efficient strategy to manage clubroot disease. However, efficiency of these resistant varieties newly integrated in low-input agricultural systems (especially low nitrogen input) needs to be evaluated. In this context, the research questions investigated in this thesis are: i) Is clubroot resistance modulation by nitrogen fertilization genotype/isolate dependent? ii) Is there an effect of nitrogen fertilization reduction on the genetic architecture of clubroot resistance in rapeseed? iii) What are the transcriptomic and metabolomic responses implicated in symptom reduction under low nitrogen fertilization? To answer these questions, clubroot responses of several rapeseed genotypes were screened using different P. brassicae isolates under two nitrogen fertilizations (8mM and 1mM of nitrate). Based on this experiment, genetic control of symptom development and resting spore production during inoculation using two isolates (eH and K92-16) was studied by linkage analyses using a bi-parental population and by genome wide association using a winter rapeseed diversity set. Last, molecular responses implicated in symptom development, observed in ‘Yudal’ genotype under 1mM condition, were studied using RNA-seq and metabolomic methods. We revealed that impact of nitrogen fertilization on symptom quantity and resting spore production are dependent to genotype/isolate combination. Modulations observed were characterized by an increase of resistance under 1mM condition. Additionally, nitrogen fertilization modulates also the effect (R²) of some Quantitative Trait Loci implicated in clubroot resistance (QTL). Using eH isolate, nitrogen fertilization principally impacts C09 QTL and C02 QTL effects. Ultimately, increase of resistance observed on ‘Yudal’ genotype under 1mM condition, using eH isolate, seems to be related with overexpression of genes implicated in nitrate assimilation (NRT2 family) and auxin transport.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-09-06T18:21:23Z No. of bitstreams: 1 2017 - Carlos Ant?nio dos Santos.pdf: 1589290 bytes, checksum: fbf16019cb63898c65e615c045d17094 (MD5) / Made available in DSpace on 2018-09-06T18:21:23Z (GMT). No. of bitstreams: 1 2017 - Carlos Ant?nio dos Santos.pdf: 1589290 bytes, checksum: fbf16019cb63898c65e615c045d17094 (MD5) Previous issue date: 2017-02-17 / CAPES / CNPq / FAPERJ / The search for efficient strategies to reduce the losses caused by clubroot (Plasmodiophora brassicae) in cauliflower is essential due to the economic and social importance of this culture. Thus, the present study was developed with the objective of evaluating the effect of different treatments in the control of the disease, root development, biomass and inflorescences production under field and greenhouse conditions. The study consisted of three trials carried out in a family farmer's area in the municipality of Nova Friburgo, RJ, Brazil. In the first trial, it was tested the residual effect of four doses of calcined limestone (0, 1.0, 2.0 and 4.0 Mg.ha-1) associated to the application of Trichoderma harzianum biofungicide under field conditions. In the second trial, it was tested the effect of the liming associated to the application of T. harzianum and cyazofamid, under greenhouse conditions. In the third one, two ways of applying limestone were tested, in the planting pit, and applied by hand. The use of gypsum was evaluated in the field. The increase of the limestone doses promoted higher pH levels, and Ca+2 contents, and reduction of the toxic Al + 3 contents in the soil. There was also a reduction of root volume with galls, greater root development, and productivity in cauliflower. The use of T. harzianum was not effective to control the disease and did not favor the root development, either in field or in greenhouse conditions. The fungicide cyazofamid reduced the volume roots with galls, the severity, and the progression of the disease. The third rial revealed a discrete reduction in the severity of the disease when using limestone and gypsum combined. Although there was no effect of the treatments on the root system, limestone applied to the pit and by hand resulted in greater accumulations of fresh mass of inflorescence and productivity. The application of limestone by hand, in the pit, and combined with gypsum favored the accumulation of calcium in the plants. The application of limestone in the pit was equivalent to the hand application in terms of soil chemical attributes improvement, reduction of disease severity, and increase of productivity and accumulation of nutrients. / A busca por estrat?gias eficientes na redu??o das perdas causadas pela h?rnia das cruc?feras (Plasmodiophora brassicae) em couve-flor ? essencial devido ? import?ncia econ?mica e social dessa cultura. Com isso, desenvolveu-se o presente estudo com o objetivo de avaliar, em condi??es de campo e casa de vegeta??o, o efeito de diferentes tratamentos no controle da doen?a, desenvolvimento das ra?zes e produ??o de biomassa e infloresc?ncias em couve-flor. O trabalho foi constitu?do de tr?s ensaios realizados em ?rea de produtor familiar no munic?pio de Nova Friburgo, RJ. No primeiro ensaio, avaliou-se o efeito residual de quatro doses de calc?rio calcinado (0; 1,0; 2,0 e 4,0 Mg.ha-1), associado ? aplica??o de biofungicida a base de Trichoderma harzianum, em condi??es de campo. No segundo, avaliou-se o efeito da calagem associada ? aplica??o de T. harzianum e ciazofamida, em casa de vegeta??o. No terceiro, avaliou-se em campo duas formas de aplica??o de calc?rio, na cova e ? lan?o, e o uso de gesso agr?cola. O aumento das doses de calc?rio promoveu eleva??o do pH e dos teores de Ca+2 e redu??o do teor de Al+3 t?xico no solo. Ainda, houve redu??o do volume de ra?zes com h?rnias e maior desenvolvimento radicular e produtividade em couve-flor. O uso de T. harzianum n?o foi eficaz no controle da doen?a e n?o favoreceu o desenvolvimento radicular das plantas, seja nas condi??es de campo ou de casa de vegeta??o. O fungicida ciazofamida reduziu o volume de ra?zes doentes, a severidade e a progress?o da doen?a. Constatou-se no terceiro ensaio discreta redu??o da severidade da doen?a com o uso de calc?rio combinado com gesso. N?o houve efeito dos tratamentos sobre o sistema radicular, entretanto, calc?rio aplicado na cova e ? lan?o resultaram em maiores ac?mulos de massa fresca de infloresc?ncia e produtividade. A aplica??o de calc?rio ? lan?o, na cova, e calc?rio combinado com gesso favoreceram o ac?mulo de c?lcio nas plantas. O uso de calc?rio na cova foi equivalente ? sua aplica??o ? lan?o quanto a melhorias dos atributos qu?micos do solo, redu??o da severidade da doen?a, aumento da produtividade e ac?mulo de nutrientes.