Les lignées clonales sont un outil puissant pour une meilleure caractérisation génétique des organismes utilisés en recherche. En utilisant les techniques de reproduction uniparentale, de telles lignées peuvent être développées chez les poissons en seulement deux générations. L'androgenèse avait déjà été réalisée chez plusieurs espèces d'eau douce et a été tentée chez le loup, une espèce d'intérêt économique et scientifique. Le faible nombre d'individus contenant effectivement uniquement le patrimoine génétique du père, après irradiation des œufs aux UV, a soulevé des questions quant à la présence de composés photoprotecteurs dans les œufs pélagiques. Les acides aminés de type mycosporines et le gadusol ont été recensés dans de nombreux organismes marins et d'eau douce, mais leur présence n'a pu être reliée au comportement flottant ou coulant des œufs. Tandis que le gadusol apparaissait en plus grande quantité dans les œufs marins flottants, ce résultat était beaucoup moins clair en eau douce et le type de composés photoprotecteurs semble lié à la phylogénie. D'autres études sur les mécanismes de photoréparation de l'ADN pourraient éclairer les résultats obtenus dans les expériences d'androgenèse. La gynogenèse avait déjà été reportée chez le loup mais le grand nombre d'individus méiotiques contaminant les descendances homozygotes produites a conduit à valider des marqueurs microsatellites efficaces à distinguer les individus mitotiques. De plus, un retard de développement des gonades a été observé, augmentant les difficultés à obtenir les lignées clonales. La grande variabilité observée entre les individus quant au succès des reproductions uniparentales a montré que la caractérisation et la qualité des gamètes sont un préalable indispensable. / Clonal lines are a powerful scientific tool for improved genetic characterization of organisms used in research. Inbred fish lines can be produced in only two generations using uniparental reproduction techniques. Androgenesis, achieved with variable success in several freshwater species, has been attempted in the European sea bass (Dicentrarchus labrax L), a marine fish of commercial and scientific interest. The low yields of progenies inheriting only the paternal genome after UV-irradiation of eggs led to considerations on the occurrence of UV screening compounds in pelagic eggs. Mycosporine-like amino acids and gadusol were found in many marine and freshwater organisms, but their occurrence in fish eggs was not clearly related to a behavioral pattern and while gadusol appeared in higher proportions in pelagic marine eggs compared to benthic species, this statement did not apply in freshwater, and moreover the kind of compounds was related to phylogeny. Further studies on DNA photorepair could enlighten hypotheses to understand the mechanisms underlying the disparate results obtained in inducing androgenesis in different fish species. Gynogenesis was reported successful to produce clonal founders in the sea bass, but high numbers of meiotic individuals contaminating fully homozygous progenies highlighted the need for efficient DNA markers to distinguish mitotic gynogenetic individuals. Furthermore, gonad development was highly delayed in gynogenetic progenies enhancing the difficulties to produce clonal lines. A high variability between individuals in the success of uniparental reproduction brought out gamete characterization and quality as a prerequisite.

Plants and microorganisms interact with each other constantly, with some interactions being mutually beneficial and others being detrimental to the plants. The features of the organisms involved in such interactions will determine the characteristics of individual pathosystems. Plants respond readily to pathogen attacks, regardless of the pathosystem; furthermore, variation in the resistance to pathogens within species is common and well documented in many plant species. The variability in pathogen resistance is at the core of genetic improvement programs for disease resistance. True resistance to pathogens in plants is a genetically determined and complex trait that can involve both constitutive and induced mechanisms at different levels of organization. The complexity of this phenomenon makes the study of compatible plant - pathogen interactions challenging, and typically, disease resistance studies focus on specific aspects of a pathosystem, such as field resistance, anatomical or physiological features of resistant plants, or molecular mechanisms of resistance. The Thuja sp. - Didymascella thujina (E.J. Durand) Maire interaction is an important pathosystem in western North America, which has been studied for more than five decades. Western redcedar (Thuja plicata Donn ex D. Don) is very susceptible to cedar leaf blight (D. thujina), a biotroph that affects the tree at all stages, although seedlings are the most sensitive to the pathogen. The characteristics of the Thuja sp. - D. thujina interaction, the wealth of information on the pathosystem and the excellent Thuja sp. genetic resources available from the British Columbia Ministry of Forests, Lands, Natural Resource Operations and Rural Development make this interaction an ideal system to advance the study of disease resistance mechanisms in conifers. This Doctoral project presents a comprehensive investigation of the constitutive and induced resistance mechanisms against D. thujina in T. plicata, Thuja standishii (Gord.) Carrière and a Thuja standishii x plicata hybrid at the phenotypic and gene expression levels, undertaken with the objective of exploring the resistance mechanisms against the biotroph in these conifers. The project also aimed to establish base knowledge for the future development of markers for marker-assisted breeding of T. plicata. The investigations included a combination of histological, chemical and next generation sequencing (NGS) methodologies. NGS data were analyzed, in addition to the traditional clustering analyses, with cutting edge machine learning methods, including grade of membership analysis, dynamic topic modelling and stability selection analysis. The studies were progressively more controlled to narrow the focus on the resistance mechanisms to D. thujina in Thuja sp. Histological characteristics related to D. thujina resistance in Thuja sp. were studied first, along with the relationship between climate of origin and disease resistance. The virulence of D. thujina was also documented early in this project. Chemical and gene expression constitutive and induced responses to D. thujina infection in T. plicata seedlings were studied next. T. plicata clonal lines were then comprehensively studied to shed light on the mechanisms behind known physiologically determined resistance. A holistic investigation of the resistance mechanisms to D. thujina in T. standishii, T. plicata and a T. standishii x plicata hybrid explored the possibility of a gene-for-gene resistance model. Thirty-five T. plicata families were screened during the four field seasons carried out between 2012 and 2015, totalling more than 1,400 seedlings scored for D. thujina severity. Thirteen of those families were used in the five studies performed during the program, along with two T. plicata seedling lines self-pollinated for five generations and three T. plicata clonal lines. One T. standishii clonal line, and one T. standishii x plicata clone were also investigated during the program. A total of 16 histological and anatomical characteristics were studied in more than 750 samples, and more than 270 foliar samples were analyzed for 60 chemical and nutritional compounds. Almost one million transcriptomic sequences in four individually assembled reference transcriptomes were examined during the program. The results of the project support the variability in the resistance to D. thujina in T. plicata, as well as the higher resistance to the pathogen in plants originating from cooler and wetter environments. The data collected also depicted the existence of age-related resistance in T. plicata, and confirmed the full resistance to the disease in T. standishii. Western redcedar plants resistant and susceptible to D. thujina showed constitutive differences at the phenotypic and gene expression levels. Resistant T. plicata seedlings had thicker cuticles, constitutively higher concentrations of sabinene, alpha-thujene, and higher levels of expression of NBS-LRR disease resistance proteins. Resistant clones of T. plicata and T. standishii had higher expression levels of bark storage proteins and of dirigent proteins. Plants from all ages, species and resistance classes studied that were infected with D. thujina showed the accumulation of aluminum in the foliage, and increased levels of sequences involved in cell wall reinforcement. Additional responses to D. thujina infection in T. plicata seedlings included the downregulation of some secondary metabolic pathways, whereas pathogenesis-related proteins were upregulated in clonal lines of T. plicata. The comprehensive approach used here to study the Thuja sp. - D. thujina pathosystem could be applied to other compatible plant-pathogen interactions. / Graduate / 2020-08-31