Les spores ou les conidies fongiques sont responsables de la dissémination des champignons filamenteux dans l'environnement (air, eau, sol,…). Ensuite les spores fongiques peuvent se déposer sur les équipements dans les ateliers de fabrication, sur les matières premières agricoles et sur les aliments. Au laboratoire, les spores sont obtenues en cultivant les champignons filamenteux en conditions optimales en termes de température, activité de l'eau, nutriments, de manière à obtenir le matériel biologique le plus rapidement possible. Or naturellement, lors de la sporulation, les champignons sont soumis à différents stress, notamment hydrique, ce qui entraîne des différences notables dans l'état physiologique de la spore. Ainsi notre objectif durant cette thèse est d’évaluer l’état physiologique des spores lorsqu’elles sont soumises à certaines conditions. Une première partie de la thèse vise à établir un nouveau modèle pour une meilleure détermination du temps de germination. L’étape suivante présente l’évaluation de l’influence du stress hydrique de la sporogénèse à la germination des spores. Les deux dernières parties présentent enfin l’évaluation des conditions de stockage sur la germination des spores. L’état physiologique est un facteur clé dans le processus de germination, il serait opportun de l’intégrer dans les modèles prédictifs de la germination. / Fungal spores or conidia are responsible for filamentous fungi spread in environment (air, water, soil…). Then, they can be found on several environments including foods. In laboratory spores are obtained under favorable conditions. However, these conditions are not real, spores are subject to various stress including water stress after their formation. These conditions can make some interactions with their physiological state. Thus, our aim consists in evaluating spores physiological state after their exposition to various conditions of storage. First part of this thesis is about definition of a new model of germination for improving germination time determination. Next step concerns evaluation of water stress during spore’s germination process. The last two parts are finally dedicated to evaluation of storage condtions on spore’s germination time. Physiological state is a key factor in the germination process. It would be appropriate to include this factor in predictive models.

N-Acylethanolamines (NAEs) are endogenous lipid metabolites that occur in a variety of dry seeds, and their levels decline rapidly during the first few hours of imbibition (Chapman et al., 1999, Plant Physiol., 120:1157-1164). Biochemical studies supported the existence of an NAE amidohydrolase activity in seeds and seedlings, and efforts were directed toward identification of DNA sequences encoding this enzyme. Mammalian tissues metabolize NAEs via an amidase enzyme designated fatty acid amide hydrolase (FAAH). Based on the characteristic amidase signature sequence in mammalian FAAH, a candidate Arabidopsis cDNA was identified and isolated by reverse transcriptase-PCR. The Arabidopsis cDNA was expressed in E. coli and the recombinant protein indeed hydrolyzed a range of NAEs to free fatty acids and ethanolamine. Kinetic parameters for the recombinant protein were consistent with those properties of the rat FAAH, supporting identification of this Arabidopsis cDNA as a FAAH homologue. Two T-DNA insertional mutant lines with disruptions in the Arabidopsis NAE amidohydrolase gene (At5g64440) were identified. The homozygous mutant seedlings were more sensitive than the wild type to exogenously applied NAE 12:0. Transgenic seedlings overexpressing the NAE amidohydrolase enzyme showed noticeably greater tolerance to NAE 12:0 than wild type seedlings. These results together provide evidence in vitro and in vivo for the molecular identification of Arabidopsis NAE amidohydrolase. Moreover, the plants with altered NAE amidohydrolase expression may provide new tools for improved understanding of the role of NAEs in germination and seedling growth.

La qualité germinative (vigueur) des semences est un caractère agronomique majeur. Elle correspond à la capacité d'un lot de semences à germer de façon rapide et homogène dans une large gamme de conditions environnementales. Cette qualité germinative est notamment contrôlée par une interaction antagoniste entre deux phytohormones, l'acide abscissique (ABA) qui induit et maintient la dormance et les gibbérellines (GAs) qui stimulent la germination et la croissance de la plantule. La dormance, qui correspond à un blocage physiologique de la germination, est un paramètre non souhaitable d'un point de vue agronomique. Par conséquent la compréhension de la régulation hormonale sur la qualité des semences représente un intérêt fort pour la communauté scientifique mais aussi pour les acteurs de la filière "semences". De nombreuses études ont démontré l'existence d’une importante régulation de la synthèse des protéines au cours de l’imbibition des graines. Cette régulation traductionnelle contribuerait à la mise en place des programmes métaboliques différents en fonction de l’état physiologique des semences pour maintenir un état de dormance ou initier le processus de germination.Le travail réalisé dans le cadre de cette thèse s’est concentré à apporter des éléments nouveaux sur le rôle de l’ABA dans la détermination de la qualité physiologique des semences au cours du développement de la graine et au cours de la germination. L’impact de l’ABA a été particulièrement décortiqué à l’aide de mutant d’Arabidopsis thaliana présentant des teneurs en ABA endogènes contrastées. Par une approche multi-omique combinant des analyses transcriptomiques, protéomiques et métabolomiques, nous avons étudié les mécanismes moléculaires et biochimiques associées avec la mise en place de la qualité physiologique des semences en relation avec l’ABA. Nos résultats ont montré qu’au-delà du contenu en ABA, l’origine tissulaire de cette hormone dans les graines gouverne de nombreux réarrangements métaboliques qui participent au déterminisme de la profondeur de dormance et de la vigueur germinative. Il apparaît un lien entre l’ABA et l’activité traductionnelle, étroitement associé au métabolisme énergétique et à l’homéostasie RedOx.L’effet de l’ABA sur l’activité traductionnelle a été suivi par une adaptation des méthodes SILAC (stable-isotope labelled amino acids in cell culture) aux grains d’Arabidopsis. Cette technique a été utilisée pour décrire la dynamique du protéome dépendante du contenu en ABA des graines au cours de leur imbibition. Nos résultats montrent que cette approche originale permet d’enrichir les connaissances sur la biologie fondamentale des semences. En effet, nous avons montré que l’ABA est un régulateur clé de la synthèse protéique dans les graines et est un contributeur majeur dans la mise en place des différents programmes traductionnels. Cette approche a montré que l’ABA exerce un contrôle sur la traduction de plus de 400 ARNm au cours de l’imbibition des graines et ouvre de nouvelles pistes pour la compréhension de la régulation de la synthèse protéique chez les semences et chez les plantes. Ces données générées offrent un nouveau regard sur le processus germinatif et de sa régulation par l’ABA.Sur la base des données existantes au laboratoire et celles générées au cours de cette thèse, nous avons également développé une utilisation de bio-marqueurs pour l’évaluation de la qualité des semences et nous avons mis au point des traitements de semences innovants. Ces technologies ont été développé en accord avec les attentes des industriels de la filière « semences ». La récente obtention d’un financement pour ce projet de recherche appliquée démontre la complémentarité des recherches effectuées au sein du laboratoire avec les besoin des industriels de la filière « semences ». / Germination vigor is a main concern in agriculture. High seed vigor is defined as the capacity of a seed lot to germinate rapidly, uniformly and in a wide range of environmental conditions. Seed quality is controlled by a dynamic balance between two antagonistic hormones, abscisic acid (ABA), which induces and maintains dormancy and gibberellins (GAs), which stimulate seed germination and seedling establishment. Seed dormancy corresponds to a block to the completion of germination and is an undesirable characteristic from an agronomic point of view. Thus, investigation of seed quality toward a better understanding of hormonal regulation is of fundamental concern for scientific community and seed industry.Recent studies have highlighted the intensive regulation of protein synthesis during seed germination. Translational regulation would govern the implementation of different metabolic programs during seed imbibition in order to maintain seed dormancy or to initiate the germination process. In this thesis, we explore the role of ABA in the control of germination quality during seed development and seed germination, using Arabidopsis thaliana mutant displaying contrasted ABA content.By combined “omic” approaches, we have highlighted the impact of ABA level on metabolic rearrangements during seed maturation. Our results showed that ABA origin in the seeds governs many metabolic rearrangements controlling dormancy depth and germination vigor. In addition, the present work suggests an intimate linkage between translational activity and ABA content, in association with energetic pathways and redox homeostasis.The impact of ABA on proteome turnover during seed germination was studied by adapting a metabolic labeling of neosynthesized proteins based on SILAC methods (stable isotope labelled amino acids in cell culture) to Arabidopsis seeds. Our results suggest that ABA is a key regulator of protein synthesis and modulates metabolic changes during seed imbibition. Indeed, this novel approach has highlighted that ABA controls the translation of more than 400 mRNAs during seed imbibition. This work provides an original perspective on the contribution of ABA and mRNA translation in seed germination and provides a valuable basis for further investigation of translational regulation in seeds and in plants.Based on existing data and those generated during this thesis, we also developed innovative seed treatments and new biomarkers for seed quality assessment. Recent funding for a maturation program dedicated to improve these biotechnologies demonstrates that our research meets the needs of seed industry.

Seeds of the root parasitic plants of the genus Orobanche germinate specifically in response to host-derived germination signals, which enables parasites to detect and attack preferred hosts. The best characterized class of germination stimulants is the strigolactones (SLs), although some species respond to non-SL compounds, such as dehydrocostus lactone (DCL). Recent work indicates that SLs are perceived by members of the KARRIKIN-INSENSITIVE2 (KAI2) gene family, and suggests that within parasitic Orobanchaceae the KAI2 genes have undergone duplication and specialization. The "diverged" clade of these genes, termed KAI2d, has been shown to bind SL germination stimulants in model system assays, but the precise role for KAI2d in regulating germination specificity in a parasitic plant has not been demonstrated. To address this issue, we used genetic and genomic approaches involving two closely related species, Orobanche cernua and O. cumana, which differ primarily in host range and stimulant preference. Orobanche cernua parasitizes tomato (and other Solanaceous crops) and responds to orobanchol, the major SL from tomato roots, whereas O. cumana specifically parasitizes sunflower and responds to DCL. Crosses between O. cernua and O. cumana produced hybrid populations that segregate for stimulant specificity, creating a tractable genetic system. Orobanche cernua contains four KAI2d genes (numbered OrceKAI2d1-4), while O. cumana contains six genes (OrcuKAI2d1-6). The DNA from 94 F2 hybrids was genotyped to identify the KAI2d gene composition and these were correlated with germination phenotype. The pattern of segregation indicated that the KAI2d genes are linked, but pointed to OrceKAI2d2 as a likely orobanchol receptor. Response to DCL was associated with inheritance of all O. cumana KAI2d genes together. Each KAI2d gene was expressed in the Arabidopsis thaliana kai2 mutant background and tested for ability to recover the mutant phenotype when exposed to SLs (including orobanchol, 5-deoxystrigol and GR24) or DCL. One O. cernua gene, OrceKAI2d2, responded to all SLs, but not DCL in this system. No DCL-specific KAI2 genes were identified. In summary, we have identified the likely SL receptor in O. cernua, and show evidence that the DCL receptor is either not a KAI2d protein, or uses KAI2d in combination with other signaling pathway components. / Ph. D. / The mechanisms by which parasitic plants of the family Orobanchaceae detect their hosts is a long-standing mystery in plant science. For over half a century it has been known that seeds of parasitic plants will lie dormant until they detect a host-derived germination stimulant. Upon perception of an appropriate germination stimulant, the parasite seeds will send out a radical that has approximately 72 hours to reach a host root before the limited nutrients within the seed are exhausted. The practical impact of this plant signaling regulation is profound, as the parasites in this family include some of the most destructive weeds in the world, including broomrapes (Orobanche and Phelipanche species) and witchweeds (Striga species). Scientists have sought to understand the signaling mechanisms in order to produce crop plants that don’t produce/exude the signal or to create chemicals that can mimic stimulants and artificially trigger parasite seed germination. Our goal was to further the understanding of the parasite germination mechanism by determining the genes involved in parasite host specificity in Orobanche, of which most members germinate in response to strigolactones (SLs). Recent work indicates that SLs are perceived by members of the KARRIKIN-INSENSITIVE2 (KAI2) gene family and suggests that within parasitic Orobanchaceae the KAI2 genes have undergone duplication and specialization. The “diverged” clade of these genes, termed KAI2d, has been shown to bind SL germination stimulants in model system assays, but the precise role for KAI2d in regulating germination specificity in a parasitic plant has not been demonstrated. To this end we used two closely related species that differ in their germination stimulant and host preferences. Orobanche cernua which like most members of Orobanchaceae responds to a SL, and O. cumana which has switched to responding to a novel germination stimulant, dehydrocostus lactone (DCL). Through genetic and genomic studies of these two species, we demonstrated that one O. cernua gene, OrceKAI2d2, responded to all SLs, but not DCL in this system. No DCL-specific KAI2 genes were identified. In summary, we have identified the likely SL receptor in O. cernua, and show evidence that the DCL receptor is either not a KAI2d protein, or uses KAI2d in combination with other signaling pathway components.

The time when the seed germinates is very important. Ability to change the time of germination may be very advantageous. It allows the emerging seed to choose the best time according to abiotic conditions, and also to avoid of competition with neighbouring individuals. The seed reacts not only on adult plants and seedlings, but also on other seeds, with which is able to communicate. For a better understanding to this mechanism I conducted an experimental study with twenty-six species. The species were selected according to their position in long succession seres of mesic/xeric abandoned fields in Český kras. The seeds were left to germinate in pairs in all combinations among them. Here I present the results where I show how the time of emergence changes, depending on presence of neighbouring seed. Also, I show how these changes in germination are related to species specific traits. Out of this, I tried to influence communication between seeds by adding activated carbon. Activated carbon should stop the communication by highly absorbing surface. The time of germination of seeds which germinated alone was not proportional to the time of germination of seeds which germinated with neighbours. This correlation showed up like nonlinear, late-emerging seeds emerging unproportionally later when emerging...

Early planting is one technique to avoid or reduce heat and drought problems that negatively affecting grain crop production. If producers adopt early planting, cold temperatures may negatively affect corn yield. It is important to select hybrids that are suited for planting earlier in the southern United States. Experiments were conducted by imposing low temperatures during seed germination. Twenty commercially available corn hybrids were evaluated for seed germination and root elongation. The first objective was: 1) To determine if some hybrids germinate better at cooler temperatures than others; and 2) Determine variation in root elongation at cold temperatures among commercially available hybrids. Corn hybrids varied significantly for seed germination and root traits under cold temperatures. Some hybrids have significantly surpassed others in seed germination traits, and they germinated earlier as well having longer radicle length. Also, there were significant differences across temperatures for all traits measured. A second objective was: 1) To quantify the effects of cold temperature on seed germination rate; 2) To evaluate the effects of different cold temperatures on seed germination behavior of corn hybrids under laboratory conditions to determine how fast they germinated; and 3) To classify hybrids for response to cold temperature using cumulative seed germination. The results showed that standard germination performance occurred at 10ºC for all hybrids, but these hybrids performed well under other cold treatments (7.2°C and 8.6° C). There were no big differences between early hybrids 93 to 105 RM (Relative Maturity) and full season 115 to 120 RM in germination % and rate in both experiments, so that means that it pretty much depends on the hybrid. Therefore, the temperature had a major influence on seed germination parameters. These findings are useful for hybrid selection with respect to cool soil temperature conditions during early planting.

Call number: LD2668 .T4 1984 E45 / Master of Science

Sustainable rehabilitation can be compromised by the inability of vegetation to survive in hostile mine wastes on a long-term basis. The adverse chemical and physical properties of mine wastes, together with extreme pH conditions and lack of nutrients, provide poor growth conditions for vegetation during seed development and germination. This raises concern for the long-term survival of vegetation through means of seed production when under stress from the punitive properties of mine wastes. Seed vigour is a function of a variety of factors to which the parent plant is subjected during seed formation and maturation. Environmental conditions experienced by the maternal plant during the growth season plays a significant role in determining subsequent germination rates in seeds. Traits of offspring seed depend on the abiotic environment attributed by the growth medium during seed development and maturation The general aim of this study was to determine the viability of seed produced by a previous generation of grass species established in eight different mine wastes and two soils (namely: gypsum wastes; gold tailings with low pyrite content; gold tailings with high pyrite content; platinum tailings; kimberlite mine waste; fluorspar mine waste; andalusite mine waste; coal discard; red soil; and vertic soil) in order to identify suitable species for specific mine wastes to ensure long-term survival through means of seed production. The species selected included: Eragrostis curvula; Eragrostis tef; Cenchrus ciliaris; Eragrostis curvula; Digitaria eriantha; Cynodon dactylon; Chloris gayana; Hyparrhenia hirta; and Sorghum bicolor. The progeny seed‟s viability and ability to germinate were determined through a pot trial study and additional germination testing at the laboratory of Advance Seed (Pty) Ltd. (AS). The germination results were correlated with the growth media analyses by statistical non-parametric correlations which indicated several significant correlations among the growth media properties themselves, and with the germination of the progeny seed. C. gayana (Rhodes grass) seed had poor germination percentages, especially seed harvested from Rhodes grass grown in acidic wastes. Seed harvested from each of the E. curvula grasses grown in various mine wastes, had excellent germination percentages. According to the Repeated Measures ANOVA statistical analysis, there was a significant influence of the growth media in which the parent grass were grown as a variable on the germination of the progeny seed batches from S. bicolor, C. ciliaris, C. gayana, and D. eriantha, indicating that the environmental factors as attributed by the growth media, i.e. the eight different mine waste materials and two soils, and experienced by the maternal plant, did indeed influence the germination of progeny seed. However, it was found that significant correlations between the properties of the growth media and the germination of the progeny seed, was species dependent. The second general aim for this study was to evaluate above-ground re-growth of parent plants after cutting in the mine waste materials and soil types mentioned above. The ability of established grasses to re-grow after a cutting event was determined by cutting the above-ground biomass of the parent grasses, after which it was scored according observable above-ground growth in the following growth season. The measurement of re-growth was subjectively done by scoring the grasses according to observable above-ground biomass. Re-growth was observed for all the perennial grass species. This can be ascribed to the grasses showing resilience to stress factors attributed by the growth media; or new grasses which emerged from seed that collected in the pots, being mistaken for re-growth; or new emerging grasses from the nodes of stolons and/or rhizomes being mistaken for re-growth. However, the emergence of new grasses was an indicator of good health, as biomass allocation to rhizomes and stolons is reduced under low nutrient availability and stress conditions. Therefore the emergence of new grasses is indicative that the plant is either tolerant to stress conditions or that the plant adapted to the restriction of growth due to the roots being bound to the size of the pot. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Sustainable rehabilitation can be compromised by the inability of vegetation to survive in hostile mine wastes on a long-term basis. The adverse chemical and physical properties of mine wastes, together with extreme pH conditions and lack of nutrients, provide poor growth conditions for vegetation during seed development and germination. This raises concern for the long-term survival of vegetation through means of seed production when under stress from the punitive properties of mine wastes. Seed vigour is a function of a variety of factors to which the parent plant is subjected during seed formation and maturation. Environmental conditions experienced by the maternal plant during the growth season plays a significant role in determining subsequent germination rates in seeds. Traits of offspring seed depend on the abiotic environment attributed by the growth medium during seed development and maturation The general aim of this study was to determine the viability of seed produced by a previous generation of grass species established in eight different mine wastes and two soils (namely: gypsum wastes; gold tailings with low pyrite content; gold tailings with high pyrite content; platinum tailings; kimberlite mine waste; fluorspar mine waste; andalusite mine waste; coal discard; red soil; and vertic soil) in order to identify suitable species for specific mine wastes to ensure long-term survival through means of seed production. The species selected included: Eragrostis curvula; Eragrostis tef; Cenchrus ciliaris; Eragrostis curvula; Digitaria eriantha; Cynodon dactylon; Chloris gayana; Hyparrhenia hirta; and Sorghum bicolor. The progeny seed‟s viability and ability to germinate were determined through a pot trial study and additional germination testing at the laboratory of Advance Seed (Pty) Ltd. (AS). The germination results were correlated with the growth media analyses by statistical non-parametric correlations which indicated several significant correlations among the growth media properties themselves, and with the germination of the progeny seed. C. gayana (Rhodes grass) seed had poor germination percentages, especially seed harvested from Rhodes grass grown in acidic wastes. Seed harvested from each of the E. curvula grasses grown in various mine wastes, had excellent germination percentages. According to the Repeated Measures ANOVA statistical analysis, there was a significant influence of the growth media in which the parent grass were grown as a variable on the germination of the progeny seed batches from S. bicolor, C. ciliaris, C. gayana, and D. eriantha, indicating that the environmental factors as attributed by the growth media, i.e. the eight different mine waste materials and two soils, and experienced by the maternal plant, did indeed influence the germination of progeny seed. However, it was found that significant correlations between the properties of the growth media and the germination of the progeny seed, was species dependent. The second general aim for this study was to evaluate above-ground re-growth of parent plants after cutting in the mine waste materials and soil types mentioned above. The ability of established grasses to re-grow after a cutting event was determined by cutting the above-ground biomass of the parent grasses, after which it was scored according observable above-ground growth in the following growth season. The measurement of re-growth was subjectively done by scoring the grasses according to observable above-ground biomass. Re-growth was observed for all the perennial grass species. This can be ascribed to the grasses showing resilience to stress factors attributed by the growth media; or new grasses which emerged from seed that collected in the pots, being mistaken for re-growth; or new emerging grasses from the nodes of stolons and/or rhizomes being mistaken for re-growth. However, the emergence of new grasses was an indicator of good health, as biomass allocation to rhizomes and stolons is reduced under low nutrient availability and stress conditions. Therefore the emergence of new grasses is indicative that the plant is either tolerant to stress conditions or that the plant adapted to the restriction of growth due to the roots being bound to the size of the pot. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Riparian vegetation is one of the most complex and abundant ecosystems in the world and it provides important ecosystem services. These services are affected by electricity production from hydropower dams. Hydropower accounts for 16% of the global electricity production and almost 50% in Sweden. One effect of hydropower is sub-daily fluctuations of water level caused by the turbines being turned on and off according to electricity demand. This is referred to as hydropeaking and has largely unknown effects on the fluvial ecosystem, and especially on the riparian vegetation. No studies have been made on the effects of hydropeaking on riparian vegetation. In this study, three native plants (Carex acuta, Betula pubescens and Salix phylicifolia x myrsinifolia) and one non-native plant (Helianthus annuus) were used as indicators (i.e., phytometers) for the effects of hydropeaking along two rivers from northern Sweden; one used for hydropower production and the other free flowing. From each of the four species, seedlings of two sizes and seeds were transplanted into five different river reaches and bank elevations along a hydropeaking gradient from none to high hydropeaking intensity. C. acuta and S. phylicifolia x myrsinifolia showed significant positive relationships to the hydropeaking gradient, likely due to their natural high tolerance to frequent inundation events. Therefore, they are suitable for restoration of river shores along reaches affected by hydropeaking. In contrast, B. pubescens was negatively related to the hydropeaking gradient, losing leaves and biomass with increasing hydropeaking intensities. It turned out to be the most sensitive species among the ones used in the experiment making it suitable as an indicator. H. annuus showed no response and therefore did not serve as impact indicator or for restoration. Germination for all native species was significantly lower along the reaches affected by hydropeaking which indicates a strong connection between hydropeaking and germination. These findings showed that recruitment becomes a bottleneck in riparian communities’ conservation along rivers affected by hydropeaking, and highlight the importance of mitigation actions focused on favoring riparian species seeds’ germination.