Thèses (M.Sc.)--Université de Sherbrooke (Canada), 1998. / Titre de l'écran-titre (visionné le 20 juin 2006). Publié aussi en version papier.

A antracnose da soja é uma doença que causa danos na cultura, principalmente na fase vegetativa. Devido aos danos causados pela doença, métodos de controle integrado estão sendo estudados. Um desses métodos é o controle biológico, no qual agentes bióticos ou seus metabólitos são usados para controlar o patógeno. Esses agentes podem atuar diretamente no patógeno ou como indutores de resistência nas plantas e, assim, contribuir para o desenvolvimento de produtos biológicos. A bactéria Streptomyces scabies produz uma toxina que pode induzir resistência em plantas, a taxtomina A. O objetivo desta pesquisa foi avaliar os efeitos diretos e indiretos da taxtomina parcialmente purificada (TPP) no controle de Colletotrichum truncatum (C. truncatum). O efeito direto da TPP na germinação de esporos e no crescimento micelial do fungo foi testado in vitro. Nos testes in vivo foram realizados a padronização do método de inoculação de C. truncatum, a escolha da cultivar a partir da reação de cultivares e o efeito de indutores na resistência sistêmica e local. Análises bioquímicas da atividade da peroxidase (POX) e teor de fenóis livres e ligados também foram conduzidos. Como resultado, observou-se que a TPP autoclavada ou não teve efeito inibitório na germinação de esporos de C. truncatum Por outro lado, a TPP não afetou o crescimento micelial do patógeno. O melhor método para inoculação de C. truncatum em soja foi via micelial, com 72 h de umidade foliar e utilizando-se o isolado CMES 1059. No teste de reação das cultivares, a DS5916IPRO foi uma das cultivares mais suscetíveis dentre as 16 testadas. No tocante a indução de resistência local, as cultivares tratadas com TPP, Bion (Acibenzolar - S - Metil) e o fungicida Priori Xtra® (azostrobina+citroconazol) apresentaram a menor severidade entre os tratamentos testados. Na avaliação da indução de resistência sistêmica, os tratamentos não diferiram (p> 0,05) do controle. As análises bioquímicas também não diferiram. Assim, pode-se concluir que a TPP na dose de 100 μg equivalentes de taxtomina/L é termoestável e tem efeito direto sob a germinação de conídios de C. truncatum. Além disso, a TPP e o ASM têm o potencial de induzir resistência local em plantas de soja contra o patógeno. / The anthracnose of soybean is a disease that causes severe damages in the crop, mainly in the vegetative phase. Due to disease damage, methods of integrated control has been studied. One of these methods is the biological control, in which a biotic agent or its metabolites are used to control a pathogen. These agents can act directly on the pathogen and/or act as resistance inducers in plants and thus contribute to the development of biological products. The bacterium Streptomyces scabies produces a toxin called thaxtomin that can induce resistance in plants. The objective of this research was to evaluate the effects of the partially purified thaxtomin (TPP) on the activation of biochemical defense mechanism in the soybean plants. Thus, the direct effect of TPP on spore germination and mycelial growth was tested in vitro. The in vivo test included standardization of inoculation of C. truncatum, cultivars reaction and the effect of inducers in the systemic and local resistance. Biochemical analyzes of peroxidase activity (POX) and content of free and bound phenols were also performed. It was observed that autoclaved or non-autoclaved TPP exhibited inhibitory effects on the germination of C. truncatum. On the other hand, TPP did not affect the mycelial growth of the pathogen. The best method for inoculation of C. truncatum was by using mycelium, with 72 h of leaf wetness and with the isolate CMES 1059. In the cultivar reaction test, it was determinate that the cultivar DS5916RR was the most susceptible out among the 16 cultivars tested. Regarding local induction resistance, the cultivars treated with TPP, Acibenzolar - S - Methyl (ASM) and fungicide Priori Xtra® exhibited the lowest severity among the treatments tested. In the evaluation of systemic resistance induction, the treatments didn\'t differ (p> 0.05) of the control. The biochemical analyses also didn\'t differ among then. Thus, it can be concluded that TPP is thermostable and has direct effect on conidium germination of C. truncatum. Besides that, the TPP and ASM have the potential to induce local resistance in soybean plants against the pathogen.

No description available.

The root system is a vital and dynamic part of a plant throughout its lifetime. Its spatial distribution is the consequence of multiple interactions with the surrounding soil medium. In particular, the presence of pathogens in soil may influence the development of the plant, especially the below-ground part, in both its physiology and its structure. Studies of diseased plant roots may take different approaches and investigate disease effects at different levels. In this study, two groups of four potato (Solanum tuberosum L.) plants were grown in middle-sized plastic pots in a greenhouse, and their root systems, together with the soil medium (i.e., sieved and autoclaved homogeneous sand), were submitted to computed tomography (CT) scanning every two weeks until 10 weeks after planting. For the "diseased" group, sand was inoculated with Streptomyces scabies EF-35, the causal agent of potato common scab, at the time of planting. Disease effects on tissue density of roots and below-ground organs, space occupancy and complexity were assessed by analyzing the CT scanning data in the spatio-temporal approach. Fluctuations of tissue density over time were different, on average, between the two groups. They were characterized by an increase of density in Week 2 for the diseased group, reflecting a hardening of tissue, and a delayed decrease relative to the "healthy" group, suggesting a slower transfer of energy from the seed potato to growing roots. Space occupancy, which was studied via volumetric growth rates evaluated from CT scan data, and complexity, which was quantified by the fractal dimension estimated from skeletonized 3-D images constructed from CT scan data, also showed differences in the first part of the experiment. Original analytical procedures based on data transformation and curve fitting in histogram analysis of CT numbers were developed to obtain those results. In conclusion, the new approach presented here, which is based on the advanced processing of CT scanning data collected over time on developing plant root systems and below-ground organs, can be recommended for future phytopathological applications.

Thesis (MSc)--University of Stellenbosch, 2000. / ENGLISH ABSTRACT: Streptomyces spp. are responsible for a large proportion of the world-wide quality deterioration of potatoes causing a potato tuber disease called cornmon scab. Determining the genetic diversity of the Streptomyces spp., especially the main pathogen, S. scabies, has been a prerequisite for the ultimate control of common scab. Techniques responsible for the classification and determination of genetic diversity have improved with advances in DNA technology. Analysis of South African (S.A.) S. scabies isolates has been focusing on the organisms' morphology, physiology, pathogenicity and melanin production, but the classification of S. scabies using DNA techniques has not yet been explored. In this study various DNA techniques were screened for optimal use in determining the genetic diversity within and among isolates of S. scabies. Bacteria had been sampled from the main potato producing regions in S.A. and a few other regions. The techniques explored included RAPDs, AFLPs, RAMS, Rep-PCR, 16S rDNA sequencing and ITS analysis. The first three techniques had to be abandoned due to non-reproducibility between the same isolate extracted on separate occasions and ITS analysis was abandoned due to sequencing difficulties. Of the three Rep-PCR techniques tested (BOX, ERIC and REP), BOX was selected because it produced the clearest and most reproducible results. BOX-PCR and 16S rDNA sequencing were therefore ultimately selected as the methods to analyse the genetic diversity of the S. scabies isolates. Information concerning the pathogenicity of the isolates was supplied by the Vegetable and Ornamental Plant Research Institute of the Agricultural Research Council (VOPI, ARC, Roodeplaat). A brief analysis of the pathogenicity prediction of the isolates in this study was explored with the PCR technique. Presence of the necJ gene was previously shown to be an indication of the pathogenicity within the Streptomyces spp. group. PCR analysis is based on the amplification of a O.72kb fragment (necl) in pathogenic isolates which was absent in non-pathogenic isolates. However, in this study the test for pathogenicity lacked specificity and sensitivity and some of the problems experienced included non-reproducibility between PCR reactions and the presence of the pathogenic fragment in the nonpathogenic isolates (as designated by VOPI, ARC). These observations led to the conclusion that this technique is not an ultimate test for pathogenicity of S. scabies isolates in a South African context. The genetic distances and similarity matrices of the Rep-PCR results were calculated using Nei's genetic distance calculation (Nei M, 1975). Clusters from these matrices were constructed using the unweighted pair group average (UPGMA) with the PAUP4 package. The clusters for the 16S rDNA sequences were formed with the Neighbor Joining (NJ) method and the PAUP4 package. The NJ trees do not take small sequencing differences into account, therefore a Parsimony Network had to be constructed. The trees obtained with the 16S rDNA sequencing techniques grouped most S. scabies isolates into one major group with a 100% bootstrap robustness of this group. More genetic diversity was illustrated by the BOX-PCR technique and the isolates were generally grouped according to their different regions of origin. However, the bootstrap values were low, indicating a lack of robustness regarding the BOXPCR clustering. This was not unexpected as the number of data points employed in the BOX technique is very limited. Both techniques revealed unexpected grouping of a few isolates. Their isolated positions could be attributed to possible misclassification or to the fact that they could be genetically different S. scabies isolates. Streptomyces spp. (other than S. scabies) displayed enough differences to place them in their own distinct groups using both techniques. Comparison of the cluster results obtained in this study did not correlate to the data supplied by the VOPI, ARC (morphology, physiology, pathogenicity and melanin production) which revealed differences between the S. scabies isolates within their respective regions. The lack of diversity displayed by the 16S rDNA technique can be attributed to the fact that only a limited section of the genome is involved making it inappropriate for intra-species genetic diversity analysis. The BOX technique takes various loci within the genome but is still not ideal for a thorough genetic diversity analysis. This study represents the first attempt to determine the genetic diversity of S. scabies in S.A. on DNA level. / AFRIKAANSE OPSOMMING: Streptomyces spp. is verantwoordelik vir 'n _groot deel van die wereld afname in aartappel kwaliteit as gevolg van die aartappelknol siekte bruinskurf. Die bepaling van die genetiese diversiteit tussen die Streptomyces spp., varal die hoof patogeen in die groep, S. scabies, is 'n vooreiste vir die uiteindelike beheer van bruinskurf. Tegnieke verantwoordelik vir die klassifikasie en bepaling van genetiese diversiteit het verbeter met vooruitgang in DNA tegnologie. Analise van Suid Afrika (S.A.) se S. scabies isolate konsentreer op die organisme se morfologie, fisiologie, patogenisiteit en malanien produksie, maar die klassifikasie van S. scabies met die behulp van DNA tegnieke is nog nie uitgevoer me. In hierdie studie is verskeie DNA tegnieke ondersoek vir optimale bepaling van genetiese diversiteit binne en tussen S. scabies isolate van S.A Bakteriee is verkry van die hoof aartappel-produserende areas in S.A. en ook van 'n paar ander areas. Die tegnieke wat in die studie gebruik is, het RAPDs, AFLPs, Rep-PKR, 16S rDNA volgordebepaling en ITS analise ingesluit. Die eersgenoemde drie tegnieke is uitgesluit as gevolg van nie-herhalende resultate tussen dieselfde isolaat geisoleer op verskillende geleenthede. ITS analise is uitgesluit as gevolg van probleme met volgordebepaling. Rep- PKR en 16S rDNA volgordebepaling is uiteindelik gekies as die mees geskikte metodes vir die analise van genetiese diversiteit tussen S. scabies isolate in hierdie studie omdat albei skynbare herhaalbare resultate gelewer het. Inligting met betrekking tot die patogenisiteit van die isolate is voorsien deur die Groente en Sierplant Instituut van die Landbou Navorsinsraad (YOPI, LNR). 'n Vinnige analise van die patogenisiteits voorspelling van die isolate is uitgevoer met die PKR tegniek. Dit is voorheen aangetoon dat die teenwoordiheid van die necJ geen dui op die patogenisiteit in die Streptomyces sp _groep. PKR analise het 'n O.72kb fragment (necl) in patogeniese isolate geamplifiseer wat nie teenwoordig was in niepatogeniese isolate nie. Hierdie toets vir patogenisiteit soos gebruik in hierdie studie was egter onspesifiek en onsensitief en sommige van die probleme wat ondervind is sluit in nie-herhaalbaarheid tussen PKR reaksies en die teenwoordigheid van die patogeniese fragment in die nie-patogeniese isolaat (soos beskryf deur YOPI, LNR). Uit hierdie waarnemings word afgelei dat die tegniek nie 'n geskikte toets vir patogenisiteit van S. scabies isolate in 'n S.A konteks is nie. Die genetiese afstande en ooreenkomstige matrikse van die Rep-PCR resultate is bereken met die genetiese afstand bepaling van Nei (Nei M, 1975). Groepe is gevorm met die "unweighted pair group average" (UPGMA) en PAUP4 pakket. Die "Neighbor Joining" (NJ) groepe is gevorm met die 16S rDNA volgordebepaling data mbv die PAUP4 pakket. Die NJ groepering neem nie klein volgorde verskille in ag nie en gevolglik moes'n "Parsimony Network" opgestel word. Die groepering met die 16S rDNA volgordebepaling het meeste van die isolate in een hoof groep geplaas met 'n 100% "bootstrap" waarde. Meer genetiese diversiteit is met die BOX-PCR tegniek gevind en isolate was oor die algemeen gegroepeer vol gens van hul oorsprong. Die "bootstrap" waardes vir die BOX tegniek was baie laag. Dit was nie onverwags nie, want die hoeveelheid data punte was beperk met die BOX tegniek. Albei tegnieke het 'n aantal afwykende isolate vertoon. Hul ge-isoleerde posisies kan toegeskryf word aan moontlike misklassifikasies van die isolaat. Die moontlikheid dat daar wel genetiese verkille tussen die isolate is, kan egter nie uitgesluit word nie. Streptomyces spp. (uitgesluit S. scabies) het genoeg variasie vertoon om hulle in hul eie groepe met die gebruik van beide tegnieke te plaas. Vergelyking tussen die groepe in die studie stem nie ooreen met die data verkry vanaf YOPl, LNR (morfologie, fisiologie, patogenesiteit en melanien produksie) nie wat verskille tussen S. scabies isolate binne 'n sekere gebied vertoon. Die gebrek aan diversiteit soos vertoon deur die 16S rDNA tegniek kan toegeskryf word aan die feit dat slegs 'n beperkte gedeelte van die genoom ondersoek word, wat dit ongeskik vir intra-species genetiese diversiteit analise maak. Die BOX tegniek neem verskeie loci in die genoom in ag, maar is steeds nie ideaal vir deeglike genetiese diversiteit analise nie. Hierdie studie verteenwoordig die eerste poging om die genetiese diversiteit van S. scabies in S.A. op DNA vlak te bepaal.

La thaxtomine A (TA) est la principale toxine produite par l'agent pathogène causant la gale commune Streptomyces scabies.La TA est nécessaire et suffisante à l'induction des symptômes de la maladie.La TA a un rôle d'inhibiteur de la synthèse de la cellulose et cause une inhibition importante de la croissance racinaire chez Arabidopsis thaliana. Ce projet avait pour objectif de caractériser un mutant d'Arabidopsis plus résistant à la TA, 9F32, dans le but de mieux comprendre le mécanisme d'action de la TA. 9F32 a démontré une résistance trois fois plus importante à la TA que Col 0, autant au niveau du phénotype global qu'au niveau de la croissance racinaire. Le taux d'entrée de la TA mesuré chez 9F32 est significativement inférieur à celui du type sauvage et comparable à celui d'un mutant présumé du transport de la toxine, txr1. Une analyse de micro-puces à ADN a montré la variation d'expression de 195 gènes chez 9F32, 174 gènes ont une expression augmentée et 21 sont réprimés (FC [supérieur ou égal] 2). Cette analyse a montré que le mutant présente une expression augmentée des réponses de stress, et plus particulièrement de la voie de réponse de défense de l'acide salicylique. 9F32 présente un phénotype et une expression transcriptionnelle partageant les caractéristiques d'une privation de phosphore, soit une petite taille et une production accrue d'anthocyanines. Finalement, l'infection du mutant 9F32 avec la bactérie S. scabies a montré une diminution des symptômes par rapport au type sauvage Col 0. Ces résultats suggèrent que la résistance à la TA procure un avantage important lors de l'infection. 9F32 est donc un outil adéquat pour l'étude du mécanisme de la TA et ses phénotypes laissent présager un nouveau mécanisme de résistance à la toxine.

The principal aim of this work was to compare chosen types of potatoes in terms of occurrence of potato scab and yield. Common scab is a wide - spread potato disease presently. Scabby marks represent the significant defect of consumption potatoes as well as seed potatoes, because they depreciate appearance and quality of potatoes. However, they don{\crq}t affect lower yields directly. The experiment was realized in a private garden. Five different types of potatoes were planted out and the experiment was repeated after two consecutive years. The experiment was evaluated partially at harvest in both years, than followed a particular statistical evaluation. On account of gained results it has been found out that the choice of potato type affects yield as well as the occurrence of potato scab.