Une analyse histopathologique et génomique d'une interaction in vitro entre Ulmus americana et Ophiostoma novo-ulmi

Authors: Aoun, Mirella
Advisor: Bernier, LouisJacobi, Volker
Abstract: Interactions between plants and fungal pathogens usually lead to the induction of different host defense mechanisms. Some of these mechanisms are the reinforcement of the plant cell wall by suberin, lignin and wall bound-phenolics, the accumulation of pathogenesis-related proteins, and the production of phytoalexins that could be toxic to the pathogen. Yet in susceptible plants, these mechanisms are not effective to produce resistance and disease develops. Dutch elm disease (DED) is a pandemic tree disease that killed millions of elm trees, especially in North America and Europe. The molecular bases of this disease are still poorly understood. With the objective of identifying genes involved in the interaction between the susceptible Ulmus americana L. and the pathogen Ophiostoma novo-ulmi Brasier, an in vitro system was developed using callus cultures inoculated with budding cells of the fungus. In order to validate the use of this system for genomic analyses of the interaction, a histopathological analysis was carried out using light and electron microscopy. Fungal colonization of the callus tissue and reactions of callus cells to the presence of the pathogen were observed at 4, 24, 48, 72 and 96 hours post-inoculation (hpi). The fungus was seen in its hyphal form by 48 hpi in all parts of the callus. Histochemical tests showed the importance of host reactions such as the accumulation of phenols, lignin, and suberin. The percentage of suberized cells was 4.6 times higher at 96 hpi in infected calli than in mock-inoculated control calli. A cDNA library using suppression subtractive hybridization (SSH) was constructed from infected elm callus tissue harvested at 72 hpi. A total of 535 expressed sequence tags were generated through partial sequencing and submitted to Genbank. These were grouped into 314 unisequences, the majority corresponding to elm genes identified during the interaction. Fifty-three unisequences representing genes involved in different pathways associated with plant defense were selected by differential screening and considered upregulated in the infected tissues. The expression profiles in mock and infected elm callus cultures of a subset of 18 elm genes were analyzed in more detail by quantitative reverse transcriptase polymerase chain reaction. These confirmed upregulation and constitutive expression of selected genes during the infection process. This study provides, for the first time, a genome-wide resource for the elm, and furthermore identifies molecular mechanisms likely involved during the interaction between U. americana and the DED pathogen.
Document Type: Thèse de doctorat
Issue Date: 2009
Open Access Date: 16 April 2018
Permalink: http://hdl.handle.net/20.500.11794/21278
Grantor: Université Laval
Collection:Thèses et mémoires

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