The placenta of the anthocerote, Dendroceros tubercularis Hatt., consists of long and branched haustorial cells, that arise from the foot and gametophyte transfer cells. Both cell types contain electron‐dense vacuolar deposits that were digested by pronase and therefore are assumed to be protein. These deposits were negative to the PATAg test for carbohydrates. Protein bodies were also found in the parenchyma cells of the foot and younger meristematic cells at the base of the capsule. Vacuolar deposits of osmiophilic material in the gametophyte cells external to the placenta were stained non‐specifically with PATAg method and were not affected by pronase. The haustorial cells have pleomorphic plastids lacking starch and a thylakoid system, whereas the transfer cells have well developed chloroplasts. No pronase‐sensitive material was detected in the apo plastic space separating gametophyte and sporophyte cells. These results suggest that protein is synthesized in the haustorial cells, perhaps from precursors provided by transfer cells, and is then transferred, via plasmodesmata, to the parenchyma cells of the foot and eventually to the cells of the growing capsule.

The placenta of two Sphagnum species was examined by electron microscopy. In contrast to all mosses so far investigated, neither sporophyte nor gametophyte placental cells of Sphagnum develop wall ingrowths. The sporophyte cells are highly vacuolate and the gametophyte cells close to them degenerate to produce a system of spaces filled with mucilage. Whether this type of placenta represents a primitive or derived condition in mosses is discussed.

Increasing habitat fragmentation and rapid global warming is changing the conditions for species populations and ecological communities around the world. This presents challenges for the maintenance of biodiversity and a dominant paradigm for conservation in fragmented habitats is given by island biogeography and metapopulation (or metacommunity) ecology. In this thesis I approach key concepts (area, connectivity and community assembly) in island biogeography and metacommunity ecology within the context of a dynamic land uplift archipelago. The presented work consists of two interwoven themes: (i) A methodological theme in which statistical approaches are developed to deal with the complexities of multispecies dynamic systems, and (ii) an applied theme dealing with community assembly and island biogeography of bryophytes on young land uplift islands. To describe island connectivity for entire species assemblages, an approach using functional principal component analysis (fPCA) on patch connectivity functions (the connectivity of an island as a continuous function of a variable representing the spatial scale of species dispersal capacities) was developed. In addition, a new statistical method, functional co-inertia analysis (fCoIA), for analyzing co-variation between multivariate species data and continuous functions was developed and applied to the relation between bryophyte species incidences and the island age/area-dynamics. Primarily asexual bryophyte species are dispersal limited and presence probabilities are related to island connectivity. No such patterns were found for species, at least occasionally, producing spores. Our results suggest that bryophyte dispersal is regulated by the contribution of spores to a regional spore rain and that bryophyte species with low spore output at the landscape level may be extra vulnerable under habitat fragmentation and loss. Having specialized asexual propagules increases the presence probabilities on islands, partly compensating for the dispersal limitation in asexual species. This suggests a trade-off between dispersal and establishment capacity, but also points to the importance of local dispersal for maintaining populations under the succession driven spatial turnover of microsites on the islands. Bryophyte colonization is strongly limited by habitat availability when a given habitats is rare, but there seems to exist a threshold over which other processes (e.g. dispersal limitation) become more important. Species with more vagile life history strategies appear to be stronger affected by the area of available habitats than many perennial species

L’algue brune Ectocarpus présente un cycle de vie haplo-diploïde avec l’alternance de deux générations multicellulaires : un gamétophyte haploïde et un sporophyte diploïde. Deux mutants présentent un changement homéotique entre les programmes de développement des générations sporophyte et gamétophyte. Les mutants réitèrent le programme de développement du gamétophyte à la place du sporophyte. Ces mutants, appelés ouroboros (oro) et samsara (sam), sont affectés dans deux gènes différents codant pour des facteurs de transcription à homéodomaine de classe TALE. Ma thèse porte sur la caractérisation des deux facteurs de transcription ORO et SAM ainsi que sur les dynamiques chromatiniennes sous-jacentes. Cette thèse présente les phénotypes des deux mutants oro et sam ainsi qu’une comparaison du transcriptome des mutants avec celui du gamétophyte et sporophyte. L’interaction entre ORO et SAM a été également testée et a lieu au niveau de chaque homéodomaine. Les préférences de liaison à l’ADN des deux facteurs de transcription ont été évaluées in vitro. Un criblage par double-hybride de levure a permis d’identifier deux sous-unités C de la famille de facteurs de transcription Nuclear Factor Y interagissant avec ORO. Cette thèse a également permis des avancées importantes dans l’étude de la régulation de la chromatine notamment en mettant au point un protocole d’immunoprécipitation de la chromatine. Ainsi, les profils de six modifications post-traductionnelles d’histones sur l’ensemble du génome ont été établis. Ce travail est pionnier dans la compréhension de la reprogrammation de la chromatine et la régulation de voies de développement majeures chez les algues brunes. / The brown alga Ectocarpus exhibits a haploid-diploid life cycle with an alternation between two multicellular generations : a haploid gametophyte and a diploid sporophyte. Two mutants exhibit homeotic switching between the sporophyte and gametophyte programs, reiterating the gametophyte program instead of switching to the sporophyte. These mutants, called ouroboros (oro) et samsara (sam), carry mutations into two different genes that code for TALE homeodomain transcription factors. This thesis aimed to characterize these two transcription factors and the chromatin dynamics associated with the alternation of generation in Ectocarpus. This thesis presents the characterisation of the oro and sam mutants and a transcriptomic comparison of the mutants with the sporophyte and gametophyte. DNA-binding preferences of the two transcription factors were evaluated using in vitro methods. ORO and SAM are able to heterodimerise via their respective homeodomains and a yeast two-hybrid screen showed that two C subunits of the Nuclear Factor Y family are able to interacting with ORO. This thesis also presents major advances in the study of chromatin regulation in the brown alga. A chromatin immunoprecipitation protocol was established and used to obtain genome-wide profiles for six histone modifications. Taken together, the data presented here suggests that ORO and SAM may be involved directly in chromatin reprogramming at generation-biased genes via an association with the NF-Y complex. The work presented represents a pioneer analysis of brown algal transcription factors and chromatin reprogramming events involved in the regulation of developmental pathways.

No description available.

Chez les angiospermes, la reproduction passe par la double fécondation. Le tube pollinique délivre deux cellules spermatiques au sein du gamétophyte femelle. Une cellule féconde la cellule œuf pour produire un zygote; l’autre féconde la cellule centrale pour produire l’endosperme. Pour assurer un succès reproductif, le développement du gamétophyte femelle au sein de l’ovule doit établir un patron cellulaire qui favorise les interactions avec le tube pollinique et les cellules spermatiques. Pour ce faire, un dialogue doit s’établir entre les différentes cellules de l’ovule lors de son développement, de même que lors de la fécondation. D’ailleurs, plusieurs types de communications intercellulaires sont supposées suite à la caractérisation de plusieurs mutants développementaux. De même, ces communications semblent persister au sein du zygote et de l’endosperme pour permettre la formation d’un embryon viable au sein de la graine. Malgré les développements récents qui ont permis de trouver des molécules de signalisation supportant les modèles d’interactions cellulaires avancés par la communauté scientifique, les voies de signalisation sont de loin très incomplètes. Dans le but de caractériser des gènes encodant des protéines de signalisation potentiellement impliqués dans la reproduction chez Solanum chacoense, l’analyse d’expression des gènes de type RALF présents dans une banque d’ESTs (Expressed Sequence Tags) spécifiques à l’ovule après fécondation a été entreprise. RALF, Rapid Alcalinization Factor, est un peptide de 5 kDa qui fait partie de la superfamille des «protéines riches en cystéines (CRPs)», dont les rôles physiologiques au sein de la plante sont multiples. Cette analyse d’expression a conduit à une analyse approfondie de ScRALF3, dont l’expression au sein de la plante se limite essentiellement à l’ovule. L’analyse de plantes transgéniques d’interférence pour le gène ScRALF3 a révélé un rôle particulier lors de la mégagamétogénèse. Les plantes transgéniques présentent des divisions mitotiques anormales qui empêchent le développement complet du sac embryonnaire. Le positionnement des noyaux, de même que la synchronisation des divisions au sein du syncytium, semblent responsables de cette perte de progression lors de la mégagamétogénèse. L’isolement du promoteur de même que l’analyse plus précise d’expression au sein de l’ovule révèle une localisation sporophytique du transcrit. La voie de signalisation de l’auxine régule également la transcription de ScRALF3. De surcroît, ScRALF3 est un peptide empruntant la voie de sécrétion médiée par le réticulum endoplasmique et l’appareil de Golgi. En somme, ScRALF3 est un important facteur facilitant la communication entre le sporophyte et le gamétophyte pour amener à maturité le sac embryonnaire. L’identification d’un orthologue potentiel chez Arabidopsis thaliana a conduit à la caractérisation de AtRALF34. L’absence de phénotype lors du développement du sac embryonnaire suggère, cependant, de la redondance génétique au sein de la grande famille des gènes de type RALF. Néanmoins, les peptides RALFs apparaissent comme d’importants régulateurs lors de la reproduction chez Solanum chacoense et Arabidopsis thaliana. / In angiosperms, reproduction occurs through double fertilization. The pollen tube delivers two sperm cells into the female gametophyte. A first sperm cell fertilizes the egg cell to produce a zygote, while the other fertilizes the central cell to produce the endosperm. To ensure reproductive success, the development of the female gametophyte within the ovule must establish a cellular pattern allowing interaction with the pollen tube and sperm cells. To this end, a dialogue must be established amongst the various cells of the ovule during its development, as well as during fertilization. Several types of communication are suggested by the analysis of developmental mutants. These communications must persist in the zygote and endosperm to allow the formation of a viable embryo within the seed. Recent developments have helped to find signaling molecules that support cell interaction models developed by the scientific community, but the signaling pathways are far from complete. In order to characterise genes encoding signaling proteins which are potentially active during reproduction in Solanum chacoense, I undertook the expression analysis of the RALF-like genes present in a bank of ESTs (Expressed Sequence Tags) specific to the ovule after fertilization. RALF, Rapid Alcalinization Factor, is a 5 kDa peptide that is part of the superfamily of Cysteine Rich Proteins (CRPs), which play a wide variety physiological roles within the plant. This expression analysis led to a detailed analysis of ScRALF3, whose expression in the plant is largely restricted to the ovule. The analysis of ScRALF3 RNAi transgenic plants revealed a function during megagametogenesis. The transgenic plants exhibit abnormal mitotic divisions that prevent the maturity of the embryo sac. The positioning of the nuclei, as well as the timing of divisions in the syncytium, appear to be responsible for the arrest of development during megagametogenesis. Isolation of the promoter as well as more accurate analysis of transcript expression reveals localisation within the ovule sporophytic tissue. The auxin signaling pathway is also involved in the regulation of ScRALF3 expression. ScRALF3 is a secreted peptide passing via the endoplasmic reticulum and the Golgi apparatus. In summary, ScRALF3 may be an important factor facilitating communication between the gametophyte and the sporophyte to allow maturation of the embryo sac. The identification of a potential orthologue in Arabidopsis thaliana led to the characterisation of AtRALF34. The lack of a phenotype during embryo sac development, however, suggests that genetic redundancy within the family of RALF-like genes is very complex. Nevertheless, the RALF peptides appear to be important regulators during reproduction in Solanum chacoense and Arabidopsis thaliana.

No description available.