Soil contamination with heavy metals is a problem of worldwide concern that is still unsolved. The analysis of the current methods of soil cleaning from heavy metals leads to conclusions that phytoremediation, i.e. soil decontamination by using plants, is one of the best methods. Although this method has not received a wide application yet and possibilities of its application are still being analysed, it is one of the most prospective soil cleaning methods due to low cost and a rather efficient cleaning of the upper surface of soil. Three kinds of Poaceae f. Species – Lolium perenne L., Poa pratensis L. and Festuca pratensis Huds. – have been chosen in this work for decontaminating soil from heavy metals. These plants were grown under artificial laboratory conditions in soil which was once and periodically contaminated with heavy metals. It was established that it is the Lolium perenne L. that most efficiently removes heavy metals and cleans soil. The results of experiments show that the selected grassy plants (perennial ryegrass, meadow-grass and fescue-grass) efficiently clean soil from heavy metals (copper, lead, manganese, zinc, nickel and chromium) when soil contains both low (0.5–120 mg/kg) and high (up to 6,850 mg/kg) concentrations of these metals. Since the perennial ryegrass absorbs the highest amount of heavy metals, the highest efficiency of soil cleaning is achieved using this plant, therefore, 80 % of the mixture of grassy plants was formed of the perennial... [to full text] / Dirvožemio tarša sunkiaisiais metalais visame pasaulyje aktuali problema iki šiol neturinti universalaus sprendimo. Išanalizavus šiuo metu dirvožemio valymui nuo sunkiųjų metalų naudojamus metodus, galima daryti išvadą, kad vienas iš tinkamiausių metodų yra fitoremediacija, t. y. dirvožemio valymas nuo teršalų panaudojant augalus. Šis metodas dar nėra plačiai taikomas ir jo taikymo galimybės dar tik tiriamos tačiau, tai vienas iš perspektyviausių dirvožemio valymo metodų dėl sąlyginai mažos kainos ir pakankamai efektyvaus paviršinio dirvožemio sluoksnio išvalymo. Atliktų eksperimentinių tyrimų rezultatai parodė, kad pasirinkta žolinė augalija – daugiametė svidrė, pievinė miglė ir tikrasis eraičinas efektyviai valo dirvožemį nuo sunkiųjų metalų (vario, švino, mangano, cinko, nikelio, chromo) tiek esant mažoms (0,5–120 mg/kg), tiek didelėms (iki 6850 mg/kg) jų koncentracijoms dirvožemyje. Kadangi daugiausiai sunkiųjų metalų iš dirvožemio sorbuoja svidrė, tai dirvožemio valymo efektyvumas naudojant šią žolinės augalijos rūšį yra efektyviausias ir sudarant žolinės augalijos mišinį 80  sudarė svidrė ir po 10  eraičinas bei miglė. Eksperimentų metu tirta, kaip sunkiųjų metalų mišinio koncentracijos kenkia hidrobiontams ir nustatyta, kad didinat sunkiųjų metalų mišinio koncentracijas (iki 0,198–5,12 g/l) jos tampa letalinėmis vaivorykštiniam upėtakiui, tuo pačiu ir kitiems vandens gyvūnams.Pagal tyrimų su vaivorykštiniais upėtakiais rezultatus nustatyta, kad stebimų... [toliau žr. visą tekstą]

Dirvožemio tarša sunkiaisiais metalais visame pasaulyje aktuali problema iki šiol neturinti universalaus sprendimo. Fitoremediacija – naujai atsirandanti ir daug žadanti technologija nestipriai užterštiems dirvožemiams valyti naudojant augalus. Šiame darbe dirvožemio valymui nuo sunkiųjų metalų pasirinktos trys žolinės augalijos rūšys – svidrė, miglė ir eraičinas, augintos modelinėmis laboratorinėmis sąlygomis vieną kartą ir periodiškai sunkiaisiais metalais teršiamuose dirvožemiuose. Nustatyta, kad geriausiai sunkiuosius metalus iš dirvožemio sorbuoja ir didžiausias dirvožemio valymo efektyvumas pasiekiamas naudojant svidrę. Augalijai sorbuojant sunkiuosius metalus iš dirvožemio dalis šių metalų (Cu, Cr, Zn, Pb, Mn, Ni) koncentruojasi žolinės augalijos šaknyse, tačiau didžioji dalis ten susikaupiančių metalų (iki 90 proc.) pereina į antžeminę augalo dalį. Didžiausios sunkiųjų metalų koncentracijos augale (iki 60 proc. ir daugiau) ir atitinkamai efektyviausias dirvožemio valymas nustatytas svidrės atveju, todėl sudarant mišinį didžiąją dalį (80 proc.) jame sudaro svidrė. Sunkiųjų metalų mišinys daro neigiamą įtaką vaivorykštinio upėtakio jauniklių fiziologiniams ir morfologiniams rodikliams, todėl didinat sunkiųjų metalų mišinio koncentracijas (iki 0,198–5,12 g/l) jos tampa letalinėmis vaivorykštiniams upėtakiams, tuo pačiu ir kitiems vandens gyvūnams. Pagal tyrimų su vaivorykštiniais upėtakiais rezultatus nustatyta, kad stebimų upėtakių, gyvenusių akvariume, užterštame 5,7... [toliau žr. visą tekstą] / Soil contamination with heavy metals is a problem of worldwide concern that is still unsolved. The analysis of the current methods of soil cleaning from heavy metals leads to conclusions that phytoremediation, i.e. soil decontamination by using plants, is one of the best methods. Although this method has not received a wide application yet and possibilities of its application are still being analysed, it is one of the most prospective soil cleaning methods due to low cost and a rather efficient cleaning of the upper surface of soil. Three kinds of Poaceae f. Species – Lolium perenne L., Poa pratensis L. and Festuca pratensis Huds. – have been chosen in this work for decontaminating soil from heavy metals. These plants were grown under artificial laboratory conditions in soil which was once and periodically contaminated with heavy metals. It was established that it is the Lolium perenne L. that most efficiently removes heavy metals and cleans soil. The results of experiments show that the selected grassy plants (perennial ryegrass, meadow-grass and fescue-grass) efficiently clean soil from heavy metals (copper, lead, manganese, zinc, nickel and chromium) when soil contains both low (0.5–120 mg/kg) and high (up to 6,850 mg/kg) concentrations of these metals. Since the perennial ryegrass absorbs the highest amount of heavy metals, the highest efficiency of soil cleaning is achieved using this plant, therefore, 80 % of the mixture of grassy plants was formed of the perennial... [to full text]

Made available in DSpace on 2016-12-08T16:24:12Z (GMT). No. of bitstreams: 1 PGCA13MA094.pdf: 3484558 bytes, checksum: 4cd098df0840598b9d19fbbc52ff65cb (MD5) Previous issue date: 2013-03-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / In regions with subtropical climates the use of grasses intercropping, mainly oats and ryegrass is a common practice, but the advantages and disadvantages of the intercrop in relation the pure culture are little studied. The aim of this work was to evaluate the production and persistence of black oat and ryegrass swards grown pure or intercropping throughout the period of use. The three treatments were randomly distributed into a complete blocks design with four replicates. The pre-grazing sward heights were 23, 20 and 17 cm for the oat, intercrop and ryegrass treatments, respectively. The post-grazing sward heights were 40% lower than their initial heights. Morphogenesis was performed using the technique of marked tillers and dynamic and tiller population density (TPD) was determined with three 10-cmdiameter PVC rings per paddock. The treatments did not differ in the final length of intact leaf, being the October month with the lowest values (approximately 48% lower). The phyllochron was not affected by the form of cultivation, but the values were increasing with the advance of the growing season. The leaf elongation rate decreased over the months, with a mean value of 0.14 cm / perfilhos.graus-day for all treatments. The advance of the growing season resulted in increases stem elongation rates, with the species in the intercrop having smaller increments in the final months. In general, rates of senescence reduced with the advance of months, and the intercrop increased tissue death of ryegrass. The intercrop TPD remained constant until October, inverting the participation of species in August. Of the tillers existing on the last grazing cycle, approximately 0, 40 and 20% were first generation of the oats, ryegrass and intercrop, respectively. Thus, despite of ryegrass pure have the lowest rate of tillering, it have the same rate of population stability of the intercrop. The pure ryegrass had the highest average net accumulation rates by offering smaller forage losses by senescence. Tiller size/density compensation was observed in the three plant communities (treatments) according to the self-thinning rule. In addition, no relationships were found when each species was analysed individually in the intercrop treatment. For the species tested the developmental stage of sward has greater influence on the morphogenesis and structural characteristics than cultivation form. The morphological similarity of oats and ryegrass enables evaluated traits similar in both forms of cultivation. When intercropping the temperate climate grasses under rotational grazing, size/density compensation mechanisms occurred throughout the grazing cycles and in the plant communities the same form of the pure cultivation to maintain the leaf area index and production capacities of the pastures / Em regiões de clima subtropical a utilização de gramíneas em consórcio, principalmente aveia e azevém, é uma prática comum, porém as vantagens e desvantagens do consórcio em relação ao cultivo puro são pouco estudadas. Objetivou-se avaliar a produção e a longevidade de pastos de aveia-preta e azevém anual cultivados puros ou em consórcio ao longo de todo o período de utilização. Os três tratamentos foram distribuídos em blocos completos casualizados com quatro repetições, tendo cada unidade experimental 98 m2. As alturas em pré-pastejo foram 23, 20 e 17 cm para a aveia, o consórcio e o azevém, respectivamente, sendo rebaixados em 40% destas alturas. A morfogênese foi realizada por meio da técnica de perfilhos marcados e a dinâmica e densidade populacional de perfilhos (DPP) em três anéis de PVC com 10 cm de diâmetro por piquete. Os tratamentos não diferiram quanto ao comprimento final de folha intacta, ficando o mês de outubro com os menores valores (aproximadamente 48% menor). Não houve efeito da forma de cultivo para os valores de filocrono, porém os valores foram crescentes com o avanço da estação de crescimento. A taxa de alongamento foliar reduziu com o passar dos meses, com valor médio de 0,14 cm/perfilho.grau-dia para todos os tratamentos. O avanço da estação de crescimento resultou em incrementos nas taxas de alongamento de colmos, com as espécies no consórcio apresentando menores incrementos nos meses finais. De maneira geral, as taxas de senescência foliar reduziram com o avanço dos meses, sendo que o consórcio aumentou a morte de tecidos do azevém. O consórcio manteve a DPP constante até outubro, apresentando uma inversão na participação das duas espécies em agosto. Dos perfilhos existentes no último ciclo de pastejo, aproximadamente 0, 40 e 20% eram da primeira geração no azevém, na aveia e no consórcio, respectivamente. Com isso, apesar de apresentar a menor taxa de aparecimento de perfilhos o azevém cultivado puro apresentou o mesmo índice de estabilidade populacional do consórcio. O azevém cultivado puro apresentou as maiores taxas de acúmulo líquido médio por apresentar as menores perdas de forragem por senescência. A compensação tamanho/densidade de perfilhos foi observada segundo a lei do auto-desbaste (self thinning rule) nos três tratamentos quando levados em conta todos os perfilhos da área, não sendo encontrada relação quando as espécies foram analisadas individualmente no consórcio. Para as espécies testadas o estádio de desenvolvimento dos pastos apresenta maior influência sobre as características morfogênicas e estruturais do que a forma de cultivo. A semelhança morfológica da aveia e do azevém possibilita características morfogênicas e estruturais parecidas em ambas as formas de cultivo. É sugestivo que em consórcios entre gramíneas de clima temperado sob lotação intermitente os mecanismos de compensação tamanho/densidade operem na comunidade de plantas e ao longo dos ciclos de pastejo da mesma forma que nos cultivos puros, mantendo elevado índice de área foliar e capacidade produtiva

La germination et la croissance hétérotrophe sont des phases clés de l'établissement des plantes. Ils sont sous contrôle génétique et affectés par la température. L'objectif de cette thèse est d'analyser la variabilité inter et intra-spécifique de cinq espèces prairiales dans leurs réponses à la température pendant la germination et la croissance hétérotrophe. Trente-deux populations de Lolium perenne, Festuca arundinacea, Dactylis glomerata, Medicago sativa, et Onobrychis viciifolia ont été évalués sous huit températures constantes entre 5 et 40°C.La nouveauté de ce travail vient de la large gamme de températures et le nombre de populations évaluées. Il a été observé que, au sein des espèces, la réaction des populations à la température indique une forte variabilité et des différences significatives. La germination n'a été pas observée à 40°C pour l'une des 23 graminées, l'O. viciifolia germé peu tandis que les variétés de M. sativa ont été peu affectées. La croissance hétérotrophe des 32 populations a été affectée par la température. La croissance des axes fortement fiable ou nulle à 40°C. Les courbes de réponse du taux de croissance maximale de l'axe sont asymétriques en forme de cloche.Des interactions significatives entre la température et la population ont été détectées pour la germination et la croissance hétérotrophe. La germination des populations des sites froids est renforcée par des températures plus chaudes et limité par les plus froids et vice-versa pour les populations chaudes adaptées. / Germination and heterotrophic growth are key phases for plant establishment. They are under genetic control and affected by temperature. The objective of this thesis was to analyse the inter- and intra-specific variability of five pasture species in their responses to temperature during germination and heterotrophic growth.Thirty two populations of Lolium perenne, Festuca arundinacea, Dactylis glomerata, Medicago sativa, and Onobrychis viciifolia were evaluated under eight constant temperatures between 5 and 40°C.The novelty of this work comes from the wide range of temperatures and the number of populations evaluated. It was observed that, within species, the response of populations to temperature shows high variability and significant differences. No germination was observed at 40°C for any of the 23 grasses, O. viciifolia germinated poorly while M. sativa varieties were little affected. Heterotrophic growth of the 32 populations was affected by temperature. The growths of the axes were negligible or lethal at 40°C. Response curves of maximum growth rate of the axis were asymmetric bell-shaped.Significant interactions between temperature and population were detected for germination and heterotrophic growth. Germination of populations from cold sites is enhanced by warmer temperatures and limited by colder ones and vice versa for warm-adapted populations. This study demonstrates that genetic variability does exist within the five studied species. This variability could be exploited to breed new varieties adapted to the future climate.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / The objective of this research was to evaluate tree pasture-based systems (PS) with elephant grass (EG) + spontaneous growing species (SGS), ryegrass (RG), for PS1; EG + SGS + forage peanut, for PS2; and EG + SGS + RG + red clover, for PS3. EG was planted 4 m spaced rows. In the cool-season, ryegrass was sowed between rows of EG; red clover was sowed and the forage peanut was preserved in respectively PS. Experimental design was completely randomized with tree treatments (SF), two replicates (paddocks) in completely split-plot time (grazing cycles). Holstein cows receiving 5.5 kg-daily and a complementary concentrate feed were used in the evaluation. The herbage mass parameters, botanical composition, forage production and stocking rate were evaluated. Samples were collected by the hand-plucking method to analyze the organic matter (OM), crude protein (CP), neutral detergent fiber (NDF) and organic matter in situ digestibility (OMISD) of pasture; phenols and tannins on legumes. Nine grazing cycle were performed during the experimental period (341 days). The mean values of forage production, stocking rate, crude protein and digestibility were 17.14; 16.80; and 19.47 t/ha; 3.28; 3.34; and 3.60 UA/ha; 13.86; 15.39; and 14.78%; 78.33; 79.23; and 79.94% on the respective PS. SGS increased significantly (P≤0,05) relation to the PS without forage legume. Similar results were observed for forage production OM an NDF in comparison to PS. Nutritive value was similar between forage legumes. Differences (P≤0,05) on condensed tannin tenors were observed between forage legumes. Considering the forage production, stocking rate and nutritive value, the PS mixed to forage legumes showed better performance. / O objetivo desta pesquisa foi avaliar três sistemas forrageiros (SF) com capim elefante (CE), espécies de crescimento espontâneo (ECE), azevém anual (AZ), como SF1; CE + ECE + AZ + amendoim forrageiro (AF), como SF2; e CE + ECE + AZ + trevo vermelho (TV), como SF3. O capim elefante foi estabelecido em linhas afastadas a cada 4 m. No período hibernal fez-se o estabelecimento do AZ entre as linhas do capim elefante; o trevo vermelho foi semeado e o amendoim forrageiro foi preservado, considerando os respectivos tratamentos. O delineamento experimental foi o inteiramente casualizado, com três tratamentos (SF), duas repetições (piquetes) em parcelas subdivididas no tempo (pastejos). Para avaliação foram usadas vacas da raça Holandesa que receberam complementação alimentar com concentrado à razão de 5,5 kg/dia. Foram avaliados parâmetros da massa de forragem pré-pastejo, a composição botânica, a produção de forragem e a lotação. Para analisar a matéria orgânica (MO), a fibra em detergente neutro (FDN), proteína bruta (PB) e digestibilidade in situ da MO (DISMO) da pastagem e os teores de fenóis e taninos das leguminosas, foram coletadas amostras de pastejo simulado. Durante o período experimental (341 dias) foram efetuados nove ciclos de pastejo. Os valores médios de produção de forragem, lotação, PB e de DISMO foram 17,14; 16,80; e 19,47 t/ha; 3,28; 3,34; e 3,60 UA/ha; 13,86; 15,39; e 14,78%; 78,33; 79,23; e 79,94% nos respectivos SF. Houve aumento significativo (P≤0,05) para espécies de crescimento espontâneo na pastagem sem leguminosa. Resultados similares foram observados entre SF para MO e FDN. O valor nutritivo foi similar entre as leguminosas forrageiras. Foram observadas diferenças (P≤0,05) entre as leguminosas para os teores de taninos condensados. Considerando a massa de forragem, a lotação e o valor nutritivo, as pastagens consorciadas com leguminosas forrageiras apresentaram melhor desempenho.

Les pollutions par les xénobiotiques, en particulier les pesticides, et les métaux lourds issus des activités agricoles présentent une grande complexité de composition chimique et de dynamique spatio-temporelle. La présence de bandes enherbées entre les parcelles cultivées et les cours d’eau permet une limitation de la diffusion de ces pollutions résiduelles vers les milieux naturels. Le compartiment végétal de ces bandes enherbées peut jouer de multiples rôles dans ce contexte de protection environnementale. L’étude comparative réalisée in situ et en conditions contrôlées de laboratoire a permis de mettre en évidence le rôle biologique du compartiment végétal avec son implication directe dans les processus in planta d’absorption, de stockage et/ou de dégradation au moins partielle. Un tel rôle phytoremédiateur est dépendant de la capacité des plantes à se maintenir sur ces milieux pollués, qui diffère selon l’espèce considérée et structure ainsi les communautés végétales des bandes enherbées. L’étude intégrative en conditions contrôlées des réponses des plantes aux interactions avec les xénobiotiques à faibles doses, à différentes échelles de complexité du fonctionnement végétal, a permis de montrer les effets de ces stress chimiques chez l’espèce modèle Arabidopsis thaliana et chez l’espèce prairiale Lolium perenne. Les xénobiotiques et les métaux lourds à des doses subtoxiques ont induit d’importants bouleversements métabolomiques et moléculaires chez ces espèces, avec des effets cryptiques de ces polluants et de leurs produits de dégradation. L’analyse en conditions de multi-pollution, qui reflètent de manière réaliste les pollutions péri-agricoles, a montré la complexité et la difficulté de prédiction des interactions entre les effets des contaminants en mélange. Ces mécanismes de réponses diffèrent selon l’espèce et le polluant et laissent supposer des divergences en termes de perception et/ou de transport des polluants, ou de coordination des réponses moléculaires et métaboliques. Arabidopsis a ainsi présenté une coordination de ses réponses orientée vers une augmentation des métabolites de stress, et une diminution des métabolites carbonés (sucres solubles), en parallèle de modifications de l’expression de gènes impliqués dans les défenses antioxydantes, les défenses contre les stress xénobiotiques, ou dans la dynamique des phytohormones. Le stress chimique a entraîné chez Lolium des modifications majeures du métabolisme azoté, ainsi qu’un remaniement des processus de photorespiration. L’analyse transcriptomique de cette espèce a de plus montré que la majorité des gènes identifiés sont impliqués dans des voies de transduction de signal, montrant ainsi la complexité des mécanismes de réponse et les couplages qui existent entre les signaux métaboliques, en particulier liés aux sucres, les voies de signalisation associées aux phytohormones, les signaux de stress et la photosynthèse. / Environmental pollutions by xenobiotics, especially by pesticides and heavy metals derived from agricultural activities, show an important complexity of chemical composition and of spatiotemporal dynamic. Vegetative filter strips between cultivated fields and streams limit the diffusion of these residual pollutions to natural environments. However, the exact biological role of plant in these buffer strips is poorly understood in this context of environmental and ecological protection. A comparative study carried out in situ and in controlled conditions highlighted the role of plant compartment in the processes of absorption, storage and/or partial degradation of pollutants in planta. Such capability of phytoremediation depends on the maintenance of a vegetal cover in area subjected to recurring flow of pesticides, it varies according to species and leads to the structuration of vegetative filter strip communities. An integrative study in controlled conditions of plant responses to low doses of pollutants allowed to analyze at different levels of complexity the impacts of chemical stresses on the model species Arabidopsis thaliana and the grassland species Lolium perenne. Low and sublethal doses of xenobiotics, associated degradation products and heavy metals induced cryptic perturbations at metabolic and molecular levels. Multi-pollution analyses, which reflect realistic conditions of environmental exposure, highlighted complex interactive effects between pollutants in mixture and the difficulty to predict them. The mechanisms of response to these chemical stresses differ according to the species and the pollutant, and suggest differences in term of perception and/or transport of pollutants, or of coordination of molecular and metabolic responses. Arabidopsis presented a coordination of its responses toward an increase of stress metabolites, a decrease of carbon metabolites (soluble carbohydrates), in parallel with modifications of gene expressions implicated on antioxidant defences, defence against xenobiotic stresses, or phytohormone dynamic. Chemical stress leads to major modifications of nitrogen metabolism in Lolium, and perturbations of processes of photorespiration. De novo transcriptomic analysis of Lolium therefore showed that a majority of identified genes are related to signal transduction pathways, highlighting the complexity of response mechanisms and the links between metabolic signals, especially linked to carbohydrate, hormonal signaling pathways, stress signals and photosynthesis. Subtoxic chemical stress induced cryptic re-engineering of plant processes that may explain the development of tolerance for some species and their persistence in area affected by residual pollution.

Les hydrocarbures aromatiques polycycliques (HAP) sont des polluants organiques persistants potentiellement mutagènes et cancérigènes. Leur transfert de l’atmosphère vers les écosystèmes, notamment vers les plantes, conditionne leur entrée dans les chaines alimentaires mais les modalités de ce transfert restent encore mal connues. L’objectif de ce travail était donc de caractériser le transfert et d’identifier les effets biologiques des HAP atmosphériques sur un système sol-plante-microorganismes symbiotiques.Un dispositif expérimental a été conçu afin de recréer en laboratoire une pollution atmosphérique par les HAP avec comme HAP modèle le phénanthrène (PHE). Le dispositif a fait l’objet d’une validation et d’une calibration élaborée de manière originale par une double approche mêlant l’expérimental à la simulation mathématique. Les niveaux d’exposition en polluant (150 µg m-3), contrôlés par des échantillonneurs passifs, se sont avérés pertinents par rapport aux conditions in situ. Ce dispositif a donc été utilisé pour exposer durant un mois des microsystèmes sol-plante-microorganismes au PHE par voie atmosphérique.Les différentes études ont mis en évidence un transfert du PHE depuis l’atmosphère vers tous les compartiments du microsystème, avec une accumulation majeure vers les feuilles de trèfle ou de ray-grass (respectivement 170 et 70 µg g־ ¹MS) et un transfert phloémien vers les racines est suggéré. Chez le trèfle, la mycorhization n’a pas été affectée, tandis que le nombre de nodules actifs a diminué de manière significative. Contrairement aux racines, la biomasse aérienne du trèfle a été significativement affectée (environ – 25%) par l’exposition au PHE atmosphérique, suggérant un impact sur le métabolisme carboné de la plante. Une expérience de marquage des trèfles au ¹³C-CO2 a effectivement montré un impact négatif du PHE atmosphérique sur la croissance, l’allocation de biomasse et l’allocation carbonée. Pour conclure, ces études ont permis non seulement de caractériser les effets biologiques et physiologiques des HAP atmosphériques sur les végétaux mais également de proposer l’utilisation du potentiel mycorhizien comme indicateur de pollution atmosphérique par les HAP. / Polycyclic aromatic hydrocarbons (PAHs) are persistent organic pollutants potentially mutagenic and carcinogenic. Transfer from the atmosphere to ecosystems, especially to plants, conditioning their entry into food chains, but the terms of this transfer are still poorly understood. The aim of this study was to characterize the transfer and identify the biological effects of atmospheric PAHs on soil-plant-symbiotic microorganisms.An experimental device was designed to recreate in the laboratory air pollution with phenanthrene (PHE) as a model PAH. The device was been validated and a calibration developed in an original way by a dual approach combining the experimental mathematical simulation. The levels of exposure to pollutant (150 mg m־³), controlled by passive samplers, were relevant with field conditions. This device has been used to expose a month of the soil-plant micro-organisms in the PHE through the air.Various studies have demonstrated a transfer of PHE from the atmosphere to all compartments of the microsystem, with a major accumulation to leaves in clover or ryegrass (respectively 170 and 70 µg g ־¹ dry weight) and a phloemic transfer to the roots is suggested. In clover, mycorrhization was not affected, while the number of active nodules decreased significantly. Unlike roots, aboveground biomass of clover was significantly affected (approximately – 25%) by exposure to air PHE, suggesting an impact on the carbon metabolism of the plant. A labelling experiment with ¹³C- CO2 in clover has actually shown a negative impact of PHE air on growth, biomass and carbon allocation.In conclusion, these studies have not only characterized the biological and physiological effects of atmospheric PAHs on plants but also proposed the use of mycorrhizal potential as an indicator of air pollution by PAHs.

No description available.

Protoplast fusion of Lolium perenne and Lotus corniculatus for gene introgression by Sanjeev V. Raikar Lolium perenne is one of the most important forage crops globally and in New Zealand. Lotus corniculatus is a dicotyledonous forage that contains valuable traits such as high levels of condensed tannins, increased digestibility, and high nitrogen fixing abilities. However, conventional breeding between these two forage crops is impossible due to their markedly different taxonomic origin. Protoplast fusion (somatic hybridisation) provides an opportunity for gene introgression between these two species. This thesis describes the somatic hybridisation, the regeneration and the molecular analysis of the putative somatic hybrid plants obtained between L. perenne and L. corniculatus. Callus and cell suspensions of different cultivars of L. perenne were established from immature embryos and plants were regenerated from the callus. Of the 10 cultivars screened, cultivars Bronsyn and Canon had the highest percentage of callus induction at 36% each on 5 mg/L 2,4-D. Removal of the palea and lemma which form the seed coat was found to increase callus induction ability of the embryos. Plant regeneration from the callus was achieved when the callus was plated on LS medium supplemented with plant growth regulators at different concentrations. Variable responses to shoot regeneration was observed between the different cultivars with the cv Kingston having the lowest frequency of shoot formation (12%). Different factors affecting the protoplast isolation of L. perenne were investigated. The highest protoplast yield of 10×106 g-1FW was obtained when cell suspensions were used as the tissue source, with enzyme combination ‘A’ (Cellulase Onozuka RS 2%, Macerozyme R-10 1%, Driselase 0.5%, Pectolyase 0.2%), for 6 h incubation period in 0.6 M mannitol. Development of microcolonies was only achieved when protoplasts were plated on nitrocellulose membrane with a L. perenne feeder layer on PEL medium. All the shoots regenerated from the protoplast-derived calli were albino shoots. The highest protoplast yield (7×106 g-1FW) of L. corniculatus was achieved from cotyledons also with enzyme combination ‘A’ (Cellulase Onozuka RS 2%, Macerozyme R-10 1%, Driselase 0.5%, Pectolyase 0.2%), for 6 h incubation period in 0.6 M mannitol. The highest plating efficiency for L. corniculatus of 1.57 % was achieved when protoplasts were plated on nitrocellulose membrane with a L. perenne feeder layer on PEL medium. The highest frequency of shoot regeneration (46%) was achieved when calli were plated on LS medium with NAA (0.1 mg/L) and BA (0.1 mg/L). Protoplast fusion between L. perenne and L. corniculatus was performed using the asymmetric somatic hybridisation technique using PEG as the fusogen. L. perenne protoplasts were treated with 0.1 mM IOA for 15 min and L. corniculatus protoplasts were treated with UV at 0.15 J/cm2 for 10 min. Various parameters affecting the fusion percentage were investigated. Successful fusions were obtained when the fusions were conducted on a plastic surface with 35% PEG (3350 MW) for 25 min duration, followed by 100 mM calcium chloride treatment for 25 min. A total of 14 putative fusion colonies were recovered. Shoots were regenerated from 8 fusion colonies. Unexpectedly, the regenerated putative hybrid plants resembled L. corniculatus plants. The flow cytometric profile of the putative somatic hybrids resembled that of L. corniculatus. Molecular analysis using SD-AFLP, SCARs and Lolium specific chloroplast microsatellite markers suggest that the putative somatic hybrids could be L. corniculatus escapes from the asymmetric protoplast fusion process. This thesis details a novel Whole Genome Amplification technique for plants using Strand Displacement Amplification technique.

Some aspects of the presence of systemic endophytic fungi in agriculturally important New Zealand grasses were studied in relation to plant breeding. Seedling resistance to adult Argentine stem weevil feeding in perennial ryegrass, Italian ryegrass and tall fescue was found to be related to the presence of their respective Acremonium endophytes in the seed rather than to plant genetic resistance. In addition a study of perennial ryegrass revealed that this resistance was independent of endophyte viability. The seedling resistance conferred by the endophyte of Italian ryegrass was found to be beneficial for field establishment. This endophyte differs from that in perennial ryegrass and tall fescue in that it does not confer resistance to Argentine stem weevil on mature plants, but only on seedlings. The extent of plant genetic seedling tolerance to adult Argentine stem weevil feeding was limited to broad inter-specific differences, with tall fescue more tolerant than perennial ryegrass and both of these more tolerant than Italian ryegrass. This ranking corresponds with previous observations on feeding preference on mature plants. A study of factors affecting the concentration of endophyte mycelia in infected seed of perennial ryegrass revealed that plant genetic factors had little effect. The major factors studied were: 1) the endophyte concentration in the maternal parent plant directly influenced the endophyte concentration in the seed. 2) nitrogen fertilizer applications to a seed crop reduced the concentration of mycelia in the seed, with earlier applications having a greater effect. 3) application of the fungicide propiconazole (Tilt) to a seed crop reduced the endophyte concentration in the seed. 4) the endophyte concentration in the seed was found to directly influence the endophyte concentration in seedlings, six month old plants and that of seed harvested from a first year seed crop. As there have been no previous reports of tetraploid perennial ryegrass cultivars with endophyte an experiment was conducted to determine if these could be developed by the standard procedure of colchicine treatment. The results revealed that endophyte was retained following colchicine treatment.