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The general aim of this thesis was to optimise the current methodologies of oocyte cryopreservation in order to obtain live offspring from cryopreserved rabbit oocytes. In chapter 1, meiotic spindle configuration, cortical granules (CGs) distribution and oocyte developmental competence were evaluated after cryopreservation with the current slow-freezing and vitrification procedures. The meiotic spindle organisation was dramatically impaired regardless of the method used. Nevertheless, altered CG distribution is more evident in vitrified oocytes than in slow-frozen ones and the developmental rate to blastocyst stage after parthenogenetic activation was only obtained using slow-freezing method. From this chapter it may be concluded that both methodologies equally affect oocyte structure. However, slow-freezing method seems to be the recommended option for this species as a consequence of the sensitivity to high levels of cryoprotectants in this species. The aim of the following two chapters was the optimisation of cryopreservation procedures using different strategies to modify the oocytes in order to make them more cryoresistant. In chapter 2, Taxol and Cytochalasin B were employed to stabilise the cytoskeleton system during vitrification. The effect of these two molecules on the meiotic spindle and chromosome configuration and development to blastocyst stage after parthenogenesis activation were also evaluated. There were no significant differences in the structural configuration between vitrified groups. Regarding cleavage and blastocyst developmental rate, no statistical differences were found between vitrified-non-treated and Taxol-treated oocytes, but no oocytes treated with Cytochalasin B reached this stage. Therefore, structural configuration and blastocyst development were not improved by this pre-treatment. Moreover, Cytochalasin B pre-treatment seems to cause a deleterious effect on developmental ability to blastocyst stage of these oocytes. In chapter 3, oocytes were incubated with cholesterol-loaded methyl-ß- cyclodextrin (CLC) to increase the membrane fluidity and stability and improve their developmental ability after parthenogenetic activation or intracytoplasmic sperm injection (ICSI). Cholesterol incorporation and its presence after cryopreservation were evaluated using confocal microscopy. Results showed that cholesterol was incorporated into the oocyte and remained, albeit in a lesser amount after cryopreservation procedures. However, no improvements on developmental competence were obtained after parthenogenetic activation or intracytoplasmic sperm injection. In the last three chapters of this thesis, the main objective was to develop a reliable technique which would allow us to obtain live offspring from cryopreserved oocytes. For that purpose, in vivo fertilisation using intraoviductal oocyte transfer assisted by laparoscopy was considered a good alternative to bypass the inadequacy of conventional in vitro fertilisation in rabbit. In chapter 4, two recipient models (ovariectomised or oviduct ligated immediately after transfer) were used to compare the ability of fresh oocytes to fertilise in vivo. This first work showed that embryo recovery rates in all transferred groups decreased significantly, but ligated oviduct recipients provided significantly higher results compared to ovariectomised ones. For that reason, in the second experiment the ligated oviduct recipient model was used to generate live births. Results obtained in this chapter suggested that it was possible to obtain offspring from cryopreserved oocytes using this technique, but this kind of animal models compromised the use of the reproductive tract in a high percentage of females. For that reason, chapter 5 was focused on the development of another type of animal model as an alternative. First, the ability of cyanoacrylate tissue adhesive to block the oviducts before the ovulation would take place was evaluated. Then, in vivo fertilisation ability of fresh transferred oocytes after blocking the oviduct with the adhesive was also assessed. Finally, slow frozen oocytes were transferred to generate live birth. Results showed that cyanoacrylate tissue adhesive was effective in blocking the oviduct, as no embryos were recovered in the blocked oviduct six days after artificial insemination (AI). Moreover, this method could fertilise fresh and also slowfrozen oocytes with a higher live birth rate than the previous recipient models. This study showed that successful production of live offspring using slow-frozen oocytes in combination with in vivo fertilisation was possible, which suggested that in vivo environment could help improve the results of oocyte cryopreservation. Thus, this method was employed in the last chapter of this thesis to generate live offspring from vitrified rabbit oocytes for the first time. Results obtained revealed that there were no differences in the rate of birth between vitrified and slow-frozen transferred oocytes. Nevertheless, based on the results with fresh oocytes, further experiments are still needed if the efficiency of cryopreservation procedures are to be improved. / Jiménez Trigos, ME. (2014). Generation of offspring from cryopreserved rabbit (Oryctolagus cuniculus) oocytes [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/37977

Yes / Plant mating systems have profound effects on levels and structuring of genetic variation and can affect the impact of natural selection. Although theory predicts that intermediate outcrossing rates may allow plants to prevent accumulation of deleterious alleles, few studies have empirically tested this prediction using genomic data. Here, we study the effect of mating system on purifying selection by conducting population-genomic analyses on whole-genome resequencing data from 38 European individuals of the arctic-alpine crucifer Arabis alpina. We find that outcrossing and mixed-mating populations maintain genetic diversity at similar levels, whereas highly self-fertilizing Scandinavian A. alpina show a strong reduction in genetic diversity, most likely as a result of a postglacial colonization bottleneck. We further find evidence for accumulation of genetic load in highly self-fertilizing populations, whereas the genome-wide impact of purifying selection does not differ greatly between mixed-mating and outcrossing populations. Our results demonstrate that intermediate levels of outcrossing may allow efficient selection against harmful alleles, whereas demographic effects can be important for relaxed purifying selection in highly selfing populations. Thus, mating system and demography shape the impact of purifying selection on genomic variation in A. alpina. These results are important for an improved understanding of the evolutionary consequences of mating system variation and the maintenance of mixed-mating strategies. / This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1707492115/-/DCSupplemental.

La chicouté présente des rendements fruitiers faibles et variables possiblement liés à l’oligotrophie des tourbières ombrotrophes où elle croît. Deux techniques de fertilisation furent donc testées dans deux tourbières naturelles, en Basse-Côte-Nord (Québec). Un engrais organo-minéral (farines de crabe et de poisson, muriate de potassium) a été appliqué au sol annuellement pendant trois ans. Des applications foliaires de bore et de cuivre ont été effectuées la troisième année. Les engrais n’ont eu aucun impact sur la croissance et la productivité de la chicouté et l’engrais organo-minéral s’est très peu minéralisé et dispersé dans le sol. La capacité de minéralisation d’autres engrais organiques en tourbière ombrotrophe devra donc être testée. Le bore et le cuivre sont relativement abondants dans les fruits et pourraient limiter la production fruitière, mais de nouveaux essais seront nécessaires pour le confirmer. En conclusion, davantage d’informations sur les besoins nutritifs de la chicouté seront nécessaires pour sélectionner les engrais appropriés. / Cloudberry present low and variable fruit yields that are probably explained by the oligotrophy of the ombrotrophic peatlands where it grows. Two different fertilization techniques were tested in two natural bogs of the Quebec Lower-North-Shore. An organo-mineral fertilizer (crab and fish meal, potassium chloride) was applied annually in soil for three years. Foliar applications of boron and copper were tested during the third year. The fertilizers had no effect on cloudberry growth and productivity and the organo-mineral fertilizer hardly mineralized and dispersed in the soil. Mineralization capacity of other organic fertilizers in natural peatlands should thus be tested. Boron and copper are abundant in the fruits and are likely limiting fruit production, but new trials are needed to confirm it. In conclusion, more information on the nutrient requirements of cloudberry is needed to select suitable fertilizers.

Thesis (PhD(Agric))--Stellenbosch University, 2009. / ENGLISH ABSTRACT: The global movement in agriculture is towards more environmentally friendly, sustainable production practices, since the role of soil microbial functions in ensuring crop production and soil fertility has become more evident in agricultural systems. Furthermore, with the impeding phase-out of methyl bromide, apple replant disease (ARD) is becoming an increasingly important problem and biological management practises are needed. Since microbial activity is generally carbon-limited in agricultural soil, it is widely accepted that management practices providing a range of organic compounds on a regular basis will tend to maintain an active and diverse microbial population. It was hypothesised that the application of various biological amendments can affect soil microbial numbers and function, thereby having a positive effect on fruit tree growth and yield. The effect of continued applications of organic material, various microbial inoculants and biostimulants on tree performance were evaluated in conventional management systems. Field trials were established in a conventional pear orchard, potential apple replant disease sites, as well as an optimally managed, high density apple orchard under controlled fertigation. The use of compost, compost extracts, a Bacillus inoculant and humates were investigated intensively. Furthermore, to improve our understanding of soil biological systems a combination of simple, practical methods were used to evaluate the effect of biological amendments on soil microbial properties and effects were related to tree performance. Regular application of compost extract in combination with compost showed the most significant effect in improving tree performance in commercial pome fruit orchards under various conditions. In the pear orchard, cumulative yield over the first two seasons was improved by more than 50% compared to controls, while in the fertigated orchard yield was improved by 22%. Biological amendments also showed improved growth in orchards suffering from stunted growth symptoms typical of ARD. However, in severe ARD cases methyl bromide fumigation showed the most consistent effects. Other biological amendments which showed positive effects on yield were application of Bacillus inoculants (Biostart®) in combination with a labile C source and a low dosage humate product, as well as a combination of compost and humates. It was clear that a combination of labile organic matter and a diverse group of microorganisms showed most promise. Although for some specific treatments increased microbial numbers and activity may have resulted in improved tree performance, in general, changes in culture-based plate counts, soil enzyme activity and carbon utilisation profiles could not be used as an indicator of yield. It was suggested that improved synchronisation of nutrient release and plant uptake, as well as microbial phytohormone production, may play an important role in improving tree performance with application of biological amendments. More research is needed on the exact mechanisms through which compost extracts improve yield and studies on root growth proliferation, as well as effects in the rhizosphere are recommended. / AFRIKAANSE OPSOMMING: Binne lanbouverband is daar tans wêreldwyd die neiging om die uitwerking van produksie-praktykte op die omgewing in ag te neem en sodoende meer verantwoordelik op te tree. Omdat die belangrike rol wat grondmikro-organisme funksionering in volhoubare verbouingspraktyke speel nou deeglik besef word, word meer volhoubare bestuurspraktyke bepleit. Hiermee saam, noodsaak aspekte soos die uitfasering van metielbromied vir die beheer van appelhervestigingsiekte, dat biologiese bestuurspraktyke meer aandag geniet. Daar word geredelik aanvaar dat gereelde toediening en aanvulling van organiese materiaal ‘n aktiewe, diverse mikrobe populasie in die grond tot gevolg sal hê. Die hipotese is gestel dat die toediening van ‘n verskeidenheid biologiese produkte grondmikrobe getalle en werking gunstig kan beïnvloed. Dit kan moontlik weer aanleiding gee tot positiewe reaskies wat die groei en drag van vrugtebome betref. In hierdie studie is die uitwerking van voortgesette toedienings van organiese materiaal, mikrobiese inokulante, asook biostimulante, op die prestasievermoë van vrugtebome ondersoek. Veldproewe is uitgelê in ‘n konvensionele peerboord, verskeie boorde met moontlike appelhervestigingsiekte probleme, asook ‘n hoëdigtheidsaanplanting appelboord onder optimale bestuur. ‘n Deeglike ondersoek is gedoen met betrekking tot die gebruik van kompos, komposekstrak, Bacillus-inokulante en humate. Eenvoudige, praktiese metodes is aangewend om vas te stel hoe biologiese toevoegings grondmikrobe eienskappe beïnvloed en of dit verband hou met veranderinge in boomprestasie. Die studie het aangetoon dat die gereelde toediening van komposekstrak saammet kompos, betekenisvolle verbetering in boomprestasie van kernvrugboorde teweeg bring onder verskeie omstandighede. Die kumulatiewe opbrengs van ‘n peerboord is oor twee seisoene met meer as 50% verhoog teenoor die kontrole. In ‘n optimaal bestuurde appelboord onder sproeibemesting, is opbrengs met 22% verhoog in vergelyking met die kontrole. Biologiese toevoegings het ook groei verbeter in boorde waar appelhervestigingsiekte bome se groei vertraag het. In die geval van ernstige appelhervestigingsimptome het metielbromied egter steeds die mees konstante positiewe uitwerking gehad. Ander biologiese toevoegings wat ‘n gunstige uitwerking op opbrengs getoon het, was ‘n kombinasie van Bacillus inokulante, ‘n lae dosis humaat en ‘n aktiewe koolstofbron, asook kompos in kombinasie met humate. Dit is duidelik dat ‘n kombinasie van ‘n maklik afbreekbare koolstofbron (soos kompos) tesame met ‘n diverse groep mikroorganismes mees belowend is vir gebruik in biologiese verbouingssisteme. Resultate toon dat veranderings in aantal organismes gemeet deur plaattellings, die aktiwiteit van grondensieme, en verbruikspotensiaal van verskillende koolstofbronne, nie as ‘n aanduiding van boomprestasie gebruik kan word nie. Daar is voorgestel dat verbeterde sinkronisasie van voedingselementvrystelling en plantopname, sowel as produksie van plantgroeihormone deur mikrobe, moontlik ‘n rol speel by boomreaksies op biologiese toevoegings. Meer navorsing wat verband hou met die meganisme waardeur komposekstrak opbrengs verbeter, is nodig. Verder word studies op fynwortelontwikkeling sowel as aspekte van die wortelrisosfeer aanbeveel.

Land-use change is the single most important global driver of changes in biodiversity. Such changes in biodiversity, in turn, are expected to influence the functioning of ecosystems and their resilience to environmental perturbations and disturbances. It is widely recognised that the use of functional traits and functional diversity is best for understanding the causes and functional consequences of changes in biodiversity, but conceptual development has outpaced empirical applications. This thesis explores these ideas in grazing systems, which are expected to undergo rapid intensification of fertiliser use and grazing pressure to meet the growing global demand for livestock products. First, a flexible framework for measuring different facets of functional diversity is described, and a new multidimensional functional diversity index, called functional dispersion (FDis), is presented. Second, two vegetation sampling methods are compared with regard to their ability to detect changes in vegetation composition. Third, shifts in plant trait distributions following land-use changes are quantified and compared to null models, and a maximum entropy approach is used to quantify the direction and strength of selection on each trait. Fourth, it is shown that these shifts in trait distributions have cascading effects on primary production, litter decomposition, soil respiration, and ultimately soil carbon sequestration. Finally, data from 18 land-use intensity gradients are used to show that land-use intensification reduces functional redundancy and response diversity, two components of biodiversity that are thought to influence ecosystem resilience to future disturbances. This study illustrates (i) the importance of considering species functional differences to understand how plant communities react to changes in soil resource availability and grazing pressure, and (ii) how such changes directly, indirectly, and interactively control ecosystem functioning, as well as (iii) increasing the vulnerability of ecosystems to future disturbances.

La fertilisation phosphatée est très répandue dans les pratiques agricoles Nord-Américaines. Bien que généralement très efficace pour augmenter la production végétale, son utilisation peut engendrer certaines contaminations environnementales. Afin de diminuer ce problème, plusieurs pratiques de gestion sont envisagées. Parmi celles-ci, on retrouve l’intéressante possibilité de manipuler la flore microbienne car cette dernière est reconnue pour son implication dans bons nombres de processus fondamentaux liés à la fertilité du sol. Cette étude a démontré que lors d’essais en champs, la forme de fertilisant ajouté au sol ainsi que la dose de phosphore (P) appliquée avaient un impact sur la distribution des microorganismes dans les différentes parcelles. Une première expérience menée sur une culture de luzerne en prairie semi-aride a montré que les échantillons provenant de parcelles ayant reçu différentes doses de P présentaient des différences significatives dans leurs communautés bactériennes et fongiques. La communauté de CMA est restée similaire entre les différents traitements. Une deuxième expérience fut menée pendant trois saisons consécutives afin de déterminer l’effet de différentes formes de fertilisation organiques et minérale ajustées selon une dose unique de P sur les populations bactériennes et fongiques d’une culture intensive de maïs en rotation avec du soja. Les résultats des analyses ont montrés que les populations varient selon le type de fertilisation reçu et que les changements sont indépendants du type de végétaux cultivé. Par contre, les populations microbiennes subissent une variation plus marquée au cours de la saison de culture. La technique de DGGE a permis d’observer les changements frappant la diversité microbienne du sol mais n’a permis d’identifier qu’une faible proportion des organismes en cause. Parallèlement à cette deuxième étude, une seconde expérience au même site fut menée sur la communauté de champignons mycorhiziens à arbuscules (CMA) puisqu’il s’agit d’organismes vivant en symbiose mutualiste avec la majorité des plantes et favorisant la nutrition de même que l’augmentation de la résistance aux stress de l’hôte. Ceci permit d’identifier et de comparer les différents CMA présents dans des échantillons de sol et de racines de maïs et soja. Contrairement aux bactéries et aux champignons en général, les CMA présentaient une diversité très stable lors des différents traitements. Par contre, au cours des trois années expérimentales, il a été noté que certains ribotypes étaient significativement plus liés au sol ou aux racines. Finalement, l’ensemble de l’étude a démontré que la fertilisation phosphatée affecte la structure des communautés microbiennes du sol dans les systèmes évalués. Cependant, lors de chaque expérience, la date d’échantillonnage jouait également un rôle prépondérant sur la distribution des organismes. Plusieurs paramètres du sol furent aussi mesurés et ils présentaient aussi une variation au cours de la saison. L’ensemble des interactions possibles entre ces différents paramètres qui, dans certains cas, variaient selon le traitement appliqué, aurait alors probablement plus d’impact sur la biodiversité microbienne que la seule fertilisation. / Phosphorus fertilization is a widespread practice in North American agriculture. Although it is generally efficient to increase yields, its use can also induce some environmental contaminations. Several management practices are considered in order to decrease this problem. Among these possibilities there is the challenging one of manipulating microbial flora, which is well known for its implication in many processes related to soil fertility. We have demonstrated in field trials that both the form of fertilizer added to soil and the applied P amounts impact microbial distribution in plots. A first experiment performed on alfalfa monocultures in semi-arid prairie conditions demonstrated that samples coming from plots that had received different doses of P fertilizer presented significant differences on their bacterial and fungal communities. AMF population remained stable between treatments. A second experiment was conducted over three growing season of an intensive maize/soybean rotation cropping system. It aimed to determine the effect of different organic and mineral fertilizers containing equal P amount on bacterial and fungal populations. It was demonstrated that these communities varied according to the fertilizer type applied. Changes are independent from the grown crop. However, microbial populations have undergone greater variation within each growing season. DGGE approach allowed to observe changes occurring in soil microbial diversity but have only permit to identify a small proportion of organisms. A second experiment in the latter study was performed on the same site and focused on arbuscular mycorrhizal fungi (AMF) as they are organisms living in a mutualistic symbiosis with most land plants and increasing host nutrition and resistance to stresses. It led to the identification and comparison of the different AMF found in maize and soybean soil and root samples. In opposition to what was observed with bacteria and fungi previously, AMF presented a very stable diversity between the different treatments. However, some ribotypes were significantly more present in soil or roots during each growing season. Finally, our whole project demonstrated that P fertilization affected microbial community structure on studied sites. Nevertheless, in each experiment, sampling time also played a substantial role in the organism distribution. Many soil parameters were also monitored and presented a seasonal variation. The sum of possible interactions between these parameters, which in some cases varied according to treatment, would thus have more impact on microbial diversity that the sole fertilization.

Le recyclage des résidus organiques sur les sols cultivés d'un territoire pourrait-il rendre possible la substitution au moins partielle des engrais azotés dans la conduite des cultures ? Cette question a été traitée dans le cas de la conduite des grandes cultures dans une zone périurbaine caractérisée par une forte diversité de résidus organiques. Le territoire étudié est la "Plaine de Versailles et le Plateau des Alluets" (PVPA), situé en Ile de France, à l'ouest de Versailles. Il représente 178km2 dont 9900ha cultivés par 82 agriculteurs dont 60 en grande culture. Il y a peu d'élevage dans la région d'étude et les sols les plus fréquents sont les luvisols et cambisols. Dans ce territoire on a mis au point une démarche d'analyse des substitutions possibles des fertilisants chimiques par des PRO (produits résiduaires organiques) qui comprend plusieurs étapes. Tout d'abord un inventaire de tous les PRO disponibles ou potentiellement disponibles sur le territoire a été réalisé et tous les PRO ont été échantillonnés et caractérisés sur un plan physicochimique : en particulier, la disponibilité du N a été évaluée via la minéralisation potentielle de l'azote organique dans des conditions contrôlées au laboratoire (norme française XPU 44-163). Les résidus organiques du territoire ont été classés en (i) PRO « amendants » (26320 tonnes de matière sèche et 442 tonnes de Ntot) caractérisés par une matière organique stabilisée, ayant un potentiel important d'entretien et d'augmentation des teneurs en matière organique des sols et (ii) les PRO « fertilisants » ayant une forte disponibilité de N (5686 tonnes de matières sèches et 361 tonnes de Ntot). Parmi les PRO recensés, les boues de stations d'épuration (séchées et chaulées), un produit commercial correspondant à un lisier de porc en provenance de Bretagne et les fientes de volailles sont les PRO ayant les valeurs fertilisantes les plus intéressantes. Des PRO à forte valeur amendante ont aussi été recensés, dont des fumiers de chevaux compostés ou non et des composts de déchets verts.[...] Suite et fin du résumé dans la thèse. / Did the recyling of organic residues of a territory on cultivated soils could make possible, at least, partial substitution of nitrogen fertilizers in crop management? This issue was treated in the case of cereal crops in a suburban area characterized by a high diversity of organic residues. The study area is the “Plaine de Versailles and the Plateau Alluets”, located in Ile de France, west of Versailles. The total surface is about 178km2 including 9900ha cultivated by 82 farmers from which 60 cultivate cereals. The animal breeding is scarce in the study area and the most common soils are Cambisols and Luvisols. We tried to develop an approach for analyzing possible substitutions of chemical fertilizers by OWP (organic waste products) use. This approach involves several steps. First an inventory of the entire OWP available or potentially available in the territory has been achieved and all OWP were sampled and characterized: in particular, the availability of N was evaluated using the potential mineralization of the organic nitrogen under controlled conditions in the laboratory (French standard XPU 44-163). Organic residues territory were classified into (i) "organic amendments" (26320 tons of dry matter and 442 tons of total N) characterized by a stabilized organic matter, with significant potential maintenance and increasing levels of soil organic matter and (ii) "organic fertilizers" with a high availability of N (5686 tons of dry matter and 361 tons of total N) . Among the identified OWP, sludge from sewage treatment (dried and limed), and a commercial product corresponding to dried pig slurry from Britain and poultry manure are OWP with the most interesting fertilizing values. Organic amendments with high stabilized organic matter value were also identified, including composted or not horse manure, and green waste compost. Last and final summary in the thesis.

This thesis focussed on the effects of a range of trace metals on various stages of Haliotis rubra development. The trace metals assessed in this thesis were the essential metals Cu, Zn and Fe; and, the non-essential metals Hg, Cd and Pb. Copper and Hg proved to be the two most toxic metals to the life stages of H. rubra studied. The concentrations affecting normal development of the fertilised egg exposed for 48h showed a decreasing order of toxicity with 48hEC50 recorded at 7µg Cu/L, 20µg Hg/L, 42µg Zn/L, 4,102µg Fe/L, 4,515µg Cd/L, and 5,111µg Pb/L. Settlement and metamorphosis occur in normal larvae when aged 5 days, and exposure of the 5 day old larvae to the metals for 48h resulted in impaired crawling success at 128µg/L Cu and Hg, and 1250µg Cd/L. Settlement was inhibited after exposure to 128µg Cu/L, 32µg Hg/L, and 1250 Cd/L. Metamorphosis of larvae 96h after exposure was inhibited by 32µg Cu/L, 512µg Zn/L, 32µg Hg/L and 625µg Cd/L. The rate of meta morphosis was enhanced after exposure to Cu and Hg at 0.5µg/L and 64 - 256µg Zn/L. Exposure to Zn at concentrations 64, 128 and 256 µg Zn/L caused an increased rate of settlement and metamorphosis. Juvenile H. rubra exposed to the six metals for 96h were most sensitive to Cu, which produced a 96hLC50 of 87µg Cu/L compared to Hg with a 96hLC50 of 173µg Hg/L. Juvenile H. rubra were relatively insensitive to Zn and Cd with the 96h LC50 of 1730µg Zn/L and 3700µg Cd/L, respectively. Exposure to individual solutions of Cu, Zn, and Cd for 28 days resulted in juvenile H. rubra bioaccumulating significant concentrations of metals in the visceramantleedible foot muscle. Accumulation of Hg was greater in the mantle-viscera-edible foot muscle. Following exposure, depuration in clean seawater for 28 days produced varying decreases in metal concentrations for each tissue compartment. Sodium-potassium activated ATPase (Na+,K+-ATPase) activity in the gills of juvenile H. rubra was significantly affected following expos ure to the trace metals for 28 days, with a decreasing order of effect on enzyme activity of Hg-Cu-Cd-Zn. Depuration of H. rubra in clean seawater for 28 days resulted in the recovery of Na+,K+-ATPase activity to varying degrees. The recovery of ATPase activity was more efficient following exposure to Cd-Zn-Cu-Hg. The overall results of this thesis provide initial baseline information to evaluate the sensitivity of H. rubra to trace metal toxicants, and these results may be utilised by regulators for establishing marine water quality guidelines to protect H. rubra and other abalone species in their natural habitats.

En Nouvelle-Calédonie, les sols développés sur roches ultramafiques (5.500 km2) sont constitués à plus de 65% d'oxy-hydroxyde de fer, ils présentent des teneurs très faibles en azote et en phosphore (1/10 à 1/100 des teneurs moyennes), de surcroît ils sont déséquilibrés pour le rapport Ca/Mg et contiennent des concentrations très élevées en métaux lourds (nickel, manganèse, cobalt, chrome). L'exploitation à ciel ouvert du nickel sur ces sols associés à des écosystèmes particulièrement originaux et diversifiés doit prendre en considération à la fois les intérêts économiques et les préoccupations écologiques. Des espèces endémiques pionnières des sols ultramafiques ou cicatricielles des sites miniers dégradés ont été identifiées. Parmi ces espèces, plusieurs espèces de Costularia (Cyperaceae) sont préconisées pour la revégétalisation des sites miniers après exploitation. Les itinéraires techniques de restauration écologique des sites miniers requièrent une approche pluridisciplinaire intégrant des données pédologiques, physiologiques et microbiologiques des écosystèmes des maquis miniers. L'analyse des relations sols/plantes sur une toposéquence comprenant différents types de maquis a permis de mettre en évidence une relation directe entre les concentrations du sol en éléments disponibles et la diversité et l'organisation des espèces, notamment pour ce qui concerne la structuration de la strate herbacée. Ce travail couplé à la mise en place par la société SIRAS Pacifique d'un champ semencier expérimental de Cyperaceae a permis de montrer que la croissance de Costularia comosa répond positivement à la fertilisation azotée et phosphatée. L'expérimentation a permis également de souligner que la vigueur, la vitesse de développement, le taux de mycorhization et la fructification des plants sont stimulés par des doses modérées de ces éléments. D'autres travaux réalisés ont apporté un approfondissement de nos connaissances sur le rôle des microorganismes associés aux racines des Costularia. Ainsi, nous avons montré que neuf espèces pionnières de Cyperaceae étudiées in situ, développent des symbioses mycorhiziennes arbusculaires. Une expérience de mycorhization contrôlée de Costularia comosa a permis de montrer que la mycorhization par Glomus etunicatum est fonctionnelle, avec une augmentation de la biomasse des plants 2,8 fois supérieure au témoin. De plus, la mycorhization réduit la concentration en nickel dans la plante, suggérant un rôle barrière du champignon mycorhizien vis-à-vis du nickel. Dans cette étude, nous présentons également une description des populations bactériennes aérobies, hétérotrophes et résistantes au nickel, isolées des racines de deux espèces de Costularia inféodées aux maquis. Ces populations bactériennes sont de faible densité et dominées par les Protéobactéries. Certaines des souches isolées présentent une tolérance à 15 mM de nickel soluble et la capacité de solubiliser les phosphates. L'avancement apporté dans les connaissances concernant la nutrition minérale des Cyperaceae et le rôle des microorganismes qui leur sont associés permet d'envisager la mise en place en routine de champs semenciers productifs et l'utilisation des microorganismes associés aux Costularia dans les itinéraires techniques de restauration écologique pour en améliorer la réussite.