The overall goal of this research was to determine what, if any, role grapevine rhizosphere bacteria play in the differing susceptibilities of New Zealand grown rootstocks to Cylindrocarpon black foot disease. The size and diversity of bacterial populations associated with the rhizospheres of grapevine rootstocks: 101-14, 5C, Schwarzmann and Riparia Gloire were evaluated. Dilution plating showed that total bacterial (P=0.012, P=0.005 for NA and KB, respectively) and fluorescent Pseudomonad (P=0.035) rhizosphere counts differed between rhizosphere and bulk soils but did not correlate with the differing susceptibilities of the rootstock varieties to black foot. No varietal differences were found for spore forming bacteria (P=0.201). SSCP banding patterns showed that species diversity was similar for most rootstocks, but that there were some differences in the composition of bacterial populations, probably attributable to vigour. Some functional characteristics of the bacteria isolated from the rhizospheres of the most and least susceptible rootstock varieties were assessed to investigate their potential to suppress the pathogen. In dual culture, bacteria from Riparia Gloire, 101-14 and the control soil all had little ability to antagonise Cylindrocarpon destructans. However, they differed in their degrees of activity for glucanase (P=0.000), protease (P=0.001) and siderophores (P=0.000). In all tests, bacterial isolates from the rhizosphere of 101-14 had the largest number of active isolates (P≤0.002); however, those from Riparia Gloire had the greatest degree of positive responses for the glucanase and siderophore assays. Bacterial isolates from the control soil produced few glucanases and no siderophores, but had the highest degree of protease activity. Bands excised and sequenced from SSCP gels frequently matched to other ‘uncultured bacteria’ in GenBank, as well as to other bacterial phyla, classes and genera commonly isolated from soil and sediment samples. These included members of the Firmicutes, Proteobacteria (α, δ, γ), Verrucomicrobia, Acidobacteria and Chromatiales. The pathogenicity of C. destructans and Fusarium oxysporum was investigated by inoculating soil containing wounded ungrafted rootstocks of 101-14, 5C, Schwarzmann and Riparia Gloire. Results indicated that F. oxysporum might be a more aggressive pathogen than C. destructans. Inoculation with F. oxysporum or C. destructans increased disease severity, P=0.018 and P=0.056, respectively at 0 cm. Rootstock variety influenced disease severity caused by C. destructans (P<0.001) and F. oxysporum (P=0.090), with rootstocks 101-14 and 5C being most susceptible to C. destructans, and Riparia Gloire and Schwarzmann most susceptible to F. oxysporum. There was also an indication that inoculation with one pathogen increased plant susceptibility to the other, with increased F. oxysporum infection in the C. destructans inoculated treatments of Riparia Gloire and Schwarzmann (P<0.05). The effect of carbohydrate stress (leaf trimming) and inoculation on C. destructans disease severity, incidence, and rootstock rhizosphere bacterial populations was evaluated by inoculating the soil containing one year old plants of Sauvignon Blanc scion wood grafted to rootstocks 101-14 and Schwarzmann. Disease severity and incidence was similar for both Schwarzmann (8.4% and 29.3%, respectively) and 101-14 (14.9% and 31.0%, respectively). When data for the moderate and no stress treatments were combined, because their effects were similar, the disease severity was significantly higher for the highly stressed plants(P=0.043). Stress did not influence disease incidence (P=0.551). Infection occurred in the non-inoculated plants, but disease severity was higher in the plants inoculated with C. destructans than those that were not. Root dry weight of highly stressed plants was lower than in both the moderately stressed (P=0.000) and unstressed plants (P=0.003). An interaction between inoculation and stress (P=0.031) showed that inoculated and highly stressed plants had the lowest root dry weight but there was no effect of rootstocks (P=0.062). There was no significant effect of carbohydrate stress (P=0.259) or inoculation (P=0.885) on shoot dry weight. SSCP banding patterns showed that bacterial diversity was generally similar between rootstocks, but stress and inoculation altered rhizosphere bacterial communities. This study has demonstrated that functionality of grapevine rhizosphere bacteria do differ between grapevine rootstock varieties that have different susceptibilities to black foot disease, but that this role needs to be further investigated if more accurate and practically relevant conclusions are to be drawn.

Speciale. / Haves kun i elektronisk udg.

[Truncated abstract] Soils of Western Australian cropping regions are very low in phosphorous. White lupin, chickpea, and faba bean are being increasingly used in rotations with wheat on these soils. Yield of wheat after a legume crop is frequently higher than its yield after wheat. It has been reported that in addition to nitrogen, legumes can also contribute to improve the availability of phosphorous for the subsequent crops. This PhD research project aimed at optimising the economic returns of wheat-legume rotations through more efficient use of P fertiliser in the legume phase as well as enhanced availability of soil P in the subsequent wheat phase

Les hydrocarbures aromatiques polycycliques (HAP) sont des polluants persistants retrouvés majoritairement dans l’environnement, qui sont reconnus comme hautement toxiques pour les organismes vivants. Les nombreux travaux réalisés sur les mécanismes contrôlant l’évolution des HAP dans le sol et dans la rhizosphère ont mis en exergue le rôle majeur de la biodégradation par les microorganismes et de la sorption aux constituants du sol. Néanmoins, un manque d’informations persiste concernant le devenir de ces HAP dans le sol, et plus particulièrement leur localisation dans les compartiments du sol ainsi que la nature et le comportement des composés issus de leur dégradation. La thèse avait donc comme principaux objectifs : (a) la localisation des HAP et des produits dérivés dans les compartiments du sol (matières organiques, phases minérales, microorganismes et plante), et (b) la quantification et l’identification des composés organiques formant les résidus liés du sol. Pour cela, nous avons couplé deux approches:(i) le suivi d’un HAP modèle marqué, le 13C-phénanthrène (13C-PHE), dans un sol artificiellement contaminé planté et non planté pour notamment déterminer par IRMS et NanoSIMS la distribution du C issu du PHE dans les compartiments et pour identifier les composés dérivés du 13C-PHE par 13C-RMN et TD/pyGCMS. (ii) le suivi d’une contamination ancienne et récente en HAP dans un sol historiquement contaminé pour étudier l’influence des matières organiques et des phases minérales (séparées par un fractionnement densimétrique) dans la rétention des HAP. Plus de 40% du C issu du 13C-PHE est retenu dans le sol après 14 jours de croissance du ray-grass. L’enrichissement en 13C observé dans la plante est du même ordre que celui du sol (E13C ≈ 0.04 % at.) démontrant une incorporation homogène du 13C-PHE entre la plante et le sol. Cet enrichissement global dans la plante est confirmé par les images NanoSIMS réalisées sur les racines. Des hotspots en 13C ont également été observés dans la rhizosphère, et à partir de leur taille, leur forme et leur composition élémentaire en C, N et S ils ont été identifiés comme étant des bactéries, ce qui met en avant leur rôle dans la dynamique des HAP du sol. Dans le sol industriel, la majorité des HAP est retrouvée dans les MO non complexées aux minéraux. En revanche, les HAP fraîchement ajoutés se sont significativement liés aux associations organo-minérales composées de quartz, de feldspaths et de phyllosilicates / Polycyclic aromatic hydrocarbons (PAHs) are major persistent pollutants in the environment that are recognized as highly toxic to living organisms. Numerous studies were carried out on the fate of PAHs in soil and in plant rhizosphere and highlighted the major contribution of biodegradation processes and of the sorption to soil constituents. However, a lack of knowledge remains about the fate of PAHs in the soil and especially their location in soil compartments as well as the nature and the behavior of degradation products. The main objectives of the thesis were: (a) to localize PAHs and their derived products within soil compartments (organic matter, minerals, microorganisms and plant), and (b) to quantify and identify the organic compounds forming bound residues. For this purpose, we used two approaches: (i) the monitoring of a labeled PAH, the 13C-phenanthrene (13C-PHE), in a spiked soil planted or not to determine by IRMS and NanoSIMS the 13C-distribution into compartments and to identify derived compounds of 13C-PHE by 13C-RMN et TD/pyGCMS. (ii) the distribution and localization of aged and freshly spiked PAH contamination in a brownfield soil to study the impact of organic matter and minerals (using soil densimetric fractionation) in the PAH retention. More than 40% of C from 13C-PHE remained in the soil after 14 days of ryegrass growth. The 13C-enrichment in plant tissue was similarly to the soil enrichment (E13C ≈ 0.04 at.%) indicating uniform uptake of 13C-PHE between the soil and plant. NanoSIMS images acquired for roots confirmed this global enrichment in the plant. 13C-Hospots were also observed in the rhizosphere, and on the basis on their size, their shape and their elemental composition in C, N and S, they were identified as bacteria, which emphasize their contribution to the fate of PAHs in soil. In the industrial soil, most of the PAHs were found attached to the free OM. However, the freshly spiked PAHs in the soil were significantly associated with organo-mineral associations containing quartz, feldspars and phyllosilicates

En utilisant le binôme Cupriavidus metallidurans CH34-Solanacées comme un modèle et comme un point de<p>départ, une étude sur l'interaction entre les plantes et les bactéries sur un substrat pollué par le cuivre a été menée<p>dans deux directions:<p>1 °) une étude en conditions de laboratoire sur les capacités de C. metallidurans CH34 à interagir avec Nicotiana<p>plumbaginifolia (les solanacées)<p>2 °) une étude sur le terrain visant à examiner les interactions entre cuprophytes et bactéries résistantes aux<p>métaux des régions minières du Katanga.<p>La première partie inclut une étude in silico visant à établir un catalogue des gènes C. metallidurans CH34<p>potentiellement impliqués par les interactions plantes-bactéries. Ce catalogue, tout en se reposant sur le génome<p>proche de Ralstonia solanacearum, bactérie phytopathogène de plusieurs espèces végétales appartenant<p>principalement à la famille des Solanaceae, il n'a pas pris en compte les orthologues des gènes clés de la<p>virulence de cette phytopathogène. Les gènes correspondants de C. metallidurans étaient situées sur les deux<p>chromosomes et ont des orthologues dans tous les génomes séquencés des Cupriavidus / Ralstonia et dans<p>Enterobacter sp. 638, endophyte de peuplier. L'étude transcriptomique, à l'aide de «microarray» a montré que<p>certains de ces gènes étaient induits, notamment des gènes impliqués dans la mobilité flagellaire (comme motA)<p>et dans la synthèse des polysaccharides extracellulaires étaient surexprimés pendant le contact entre les plantes et<p>les bactéries, tandis que phcA (impliqué dans la détection de la densité de population et dans la conversion<p>phénotypique) et des gènes impliqués dans la biosynthèse de pili étaient sousexprimés dans les conditions<p>expérimentales testées. En outre, le contact avec les plantes semble avoir induit la surexpression des gènes<p>impliqués dans la réponse de cuivre et d'autres métaux. La capacité de C. metallidurans CH34 à coloniser<p>l'endosphere de N. plumbaginifolia a été confirmée in vitro ainsi qu'un effet de promotion de la croissance des<p>plantes dans certaines conditions. Mais la densité de la colonisation (104-106 c.f.u/g. poids frais) est<p>considérablement réduite dans des conditions non stériles et en l'absence de pression de sélection métallique.<p>La deuxième partie de l'étude s'est concentrée sur la microbiologie de cuprophytes (Haumaniastrum katagense et<p>Crepidorhopalon tenuis) dans l'arc cuprifère du Katanga: des isolats Cuprorésistants appartenant aux genres<p>Stenotrophomonas et Sphingomonas prédominent dans la rhizosphère alors que des isolats appartenant aux<p>genres Methylobacterium, Xanthomonas et Variovorax prédominent dans l'endosphere. Certaines de ces<p>bactéries sont plus résistantes au Cu(II), à des concentrations minimales inhibitrices (MIC) allant jusqu'à 5 mM,<p>que C. metallidurans CH34 (MIC: 1,5 mM) et la plupart d'entre elles résistent également aux Zn(II), Co(II) et<p>Cd(II). Des isolats appartenant au genre Cupriavidus/Ralstonia ont été détectés dans la rhizosphère des<p>cuprophytes ainsi que les séquences 16S rDNA de C. metallidurans ont été également détectées dans l'ADN<p>total extrait des cuprophytes. La détection via la réaction de la polymérase en chaîne (PCR) de gènes de<p>résistance au cuivre correspondant à des protéines periplasmiques a confirmé la présence dans les bactéries<p>cuprorésistantes, principalement de copA et dans une moindre mesure celle de copK. Mais les gènes homologues<p>de copA et de copK n'ont pas été détectés dans tous les bactéries du genre Methylobacterium dont les membres<p>ont été pourtant les plus résistants aux métaux. Certaines bactéries isolées sont capables d'interagir avec le<p>système hormonal végétal et quelques unes semblent également manifester un effet de promotion de la<p>croissance des plantes. Les premières tentatives d'élaboration de protocoles de reinoculation des bactéries<p>endophytic cuprorésistantes dans Haumaniastrum katagense ont été effectués. La biologie moléculaire et<p>l'écologie des interactions plantes-bactéries et des mécanismes de résistance métallique décrits dans ce travail<p>peuvent préparer la voie à de nouvelles applications en bioremédiation (phytostabilization / phytoextraction de<p>métaux toxiques). / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

Genetically modified (GM) crops are commercially cultivated worldwide but there are concerns on their possible negative impacts on soil biodiversity. A glasshouse study was conducted to determine effects of Bt maize residues on soil microbial diversity. Residues of Bt maize (PAN 6Q-308B) and non-Bt maize (PAN 6Q-121) were incorporated into the soil and corresponding maize seeds planted. The treatments were replicated three times. Fertilizer and water application were similar for both treatments. Rhizosphere and bulk soil was destructively sampled from each treatment and analyzed for microbial community level physiological profiles using Biolog plates with 31 different carbon substrates. Absorbance in the Biolog plates was recorded after 72 h of incubation at 20oC. Arbuscular mycorrhizal fungi spore counts were also determined. Field studies were conducted at the University of Free State and University of Fort Hare Research Farms to determine the effects of growing Bt maize on soil microbial diversity. One Bt maize cultivar (PAN6Q-308B) and non-Bt maize (PAN6Q-121) were grown in a paired experiment at University of Free State farm, while two Bt maize (DKC61-25B and PAN6Q-321B) and their near-isogenic non-Bt maize lines (DKC61-24 and PAN6777) were grown in a randomized complete block design with three replicates. Fertilization, weed control and water application, were similar for both Bt maize cultivars and their non-Bt maize counterparts. Rhizosphere soil samples were collected by uprooting whole plants and collecting the soil attached to the roots. The samples were analysed for microbial diversity and for arbuscular mycorrhizae fungal spore counts. Principal component analysis showed that soil microbial diversity was affected more by sampling time whereas genetic modification had minimal effects. Presence of residues also increased the diversity of microorganisms. Mycorrhizal fungal spores were not affected by the presence of Bt maize residues. Growing Bt maize had no effect on the soil microbial diversity in the rhizosphere.

Ce projet explore l’hypothèse selon laquelle les gènes et les métabolites végétaux communément régulés joueraient un rôle majeur dans l’interaction riz-PGPR et constituerait une signature moléculaire de la perception des PGPR par le riz. Dans cet objectif, une analyse intégrant le suivi de l’expression d’une sélection de gènes ainsi que le profilage des métabolites secondaires a été conduite sur les racines d’un unique cultivar de riz (Nipponbare) en réponse à l’inoculation de dix souches de PGPR appartenant à divers genres bactériens (Azospirillum, Herbaspirillum, Paraburkholderia). Nos résultats ont permis l’identification de quatre gènes de riz pouvant être considérés comme marqueurs de l’association riz-PGPR, avec notamment deux gènes impliqués dans la biosynthèse de phytoalexines et un gène codant pour une protéine PR (pathogenesis-related). De plus, une signature métabolique commune, constituée de neuf composés, a été mise en évidence, dont la réduction de l’accumulation de trois alkylrésorcinols et l’augmentation de l’accumulation de deux amides d’acides hydroxycinnamiques (HCAA) : la N-p-coumaroylputrescine et la N-féruloylputrescine. Cette signature métabolique a été corrélée avec l’augmentation de l’expression de deux gènes impliqués dans la biosynthèse de la N-féruloylputrescine. Il est intéressant d’observer que la confrontation du riz à un pathogène bactérien entraine une réduction de l’accumulation de ces HCAA dans les racines. Cette accumulation d’HCAA, qui sont des composés antimicrobiens potentiels, pourrait être considérée comme une réaction primaire de la perception de bactéries par le riz / Besides, a common metabolomic signature of nine compounds was highlighted, with the reduced accumulation of three alkylresorcinols and increased accumulation of two hydroxycinnamic acid amides (HCAA), identified as N-p-coumaroylputrescine and N-feruloylputrescine. This coincided with the increased transcription of two genes involved in the N-feruloylputrescine biosynthetic pathway. Interestingly, exposure to a rice bacterial pathogen triggered a reduced accumulation of these HCAA in roots. Accumulation of HCAA, that are potential antimicrobial compounds, might be considered as a primary reaction of rice to bacterial perception

Le microbiome de la rhizosphère fait généralement référence aux communautés bactériennes, archées et fongiques ainsi qu'à leur matériel génétique entourant étroitement les systèmes racinaires des plantes. Le métagénome de ce microbiome a été appelé le deuxième génome de la plante puisqu’elle est capable de profiter de plusieurs fonctions dont elle manque. La communauté microbienne de la rhizosphère inclue entre autres des microorganismes ayant développé des interactions intimes et spécifiques de longue durée avec les racines des plantes. Il s'agit d'une communauté dynamique de microorganismes, à partir de laquelle une partie d’espèces a développé des interactions intimes et spécifiques de longue durée avec les racines des plantes. Les progrès récents dans l’étude des interactions plantes-microbes ont démontré leur impact considérable sur la croissance, la nutrition et la santé des plantes. Le microbiote de la rhizosphère est complexe avec une structure spatio-temporelle dynamique qui s'adapte rapidement en fonction des stress biotiques et abiotiques. Considérant l’importance du microbiome de la rhizosphère pour la santé des plantes, des informations précises sur leurs microbes associés sont d'une importance capitale pour déchiffrer les mécanismes d'adaptation des plantes aux stress médiés par le microbiome et comprendre comment les plantes recrutent des taxons microbiens clés pour mieux faire face aux conditions stressantes. Pour ce faire, nous avons mené trois études afin de faire la lumière sur les facteurs qui jouent un rôle dans le recrutement et la structure du microbiome de la rhizosphère de plantes dans les milieux stressés. Dans un premier lieu, nous avons testé si des inoculations répétées avec des protéobactéries influençaient la productivité des plantes et les communautés microbiennes associées à la rhizosphère de quatre espèces végétales poussant dans des sédiments contaminés par des hydrocarbures pétroliers. Une expérience de mésocosme a été réalisée en conception de blocs randomisés avec deux facteurs : 1) la présence ou l'absence de quatre espèces végétales collectées dans un bassin de sédimentation d'une ancienne usine pétrochimique, et 2) l'inoculation ou non avec un consortium bactérien composé de dix isolats de Protéobactéries. Les plantes ont été cultivées en serre pendant quatre mois. Le séquençage d'amplicon MiSeq, ciblant le gène de l'ARNr 16S bactérien l’ITS fongique, a été utilisé pour évaluer les structures de la communauté microbienne des sédiments provenant de mesocosmes plantés ou non plantés. Nos résultats ont montré qu’alors que l'inoculation provoquait un changement significatif dans les communautés microbiennes, la présence de la plante et de son identité spécifique avait une influence plus forte sur la structure du microbiome dans les sédiments contaminés par les hydrocarbures pétroliers. Ensuite, en utilisant le même dispositif expérimental, nous avons utilisé le séquençage d'amplicon MiSeq ciblant le gène de l'ARNr 18S pour évaluer les structures communautaires AMF dans les racines et la rhizosphère de plantes poussant dans des substrats contaminés et non contaminés. Nous avons également étudié la contribution de l'identité spécifique des plantes et du biotope (racines des plantes et sol rhizosphérique) dans la formation des assemblages AMF associés. Nos résultats ont montré que si l'inoculation provoquait un changement significatif dans les communautés AMF, la contamination du substrat avait une influence beaucoup plus forte sur leur structure, suivie par le biotope et l'identité végétale dans une moindre mesure. De plus, l'inoculation augmentait considérablement la production de biomasse végétale et était associée à une diminution de la dissipation des hydrocarbures pétroliers dans le sol contaminé. Le résultat de cette étude fournit des connaissances sur les facteurs influençant la diversité et la structure communautaire de l'AMF associée aux plantes en milieux stressés à la suite d’inoculations répétées d'un consortium bactérien. Finalement, nous avons testé l’effet d’une inoculation d’arbres avec des champignons mycorhiziens spécifiques sur leur survie et croissance, ainsi que l’extraction de métaux traces. Pour ce faire, une expérience sur le terrain a été menée dans laquelle nous avons cultivé le clone de Salix miyabeana "SX67" sur le site d'une décharge industrielle déclassée, et inoculé les arbustes avec le champignon arbusculaire mycorhizien Rhizophagus irregularis, le champignon ectomycorhizien Sphaerosporella brunnea, ou un mélange des deux. Après deux saisons de croissance, les saules inoculés avec le champignon S. brunnea ont produit une biomasse significativement plus élevée. Le Ba, le Cd et le Zn se sont avérés être accumulés dans les parties aériennes des plantes, où le Cd présentait les valeurs de facteur de bioconcentration les plus élevées dans tous les traitements. De plus, les parcelles où les saules ont reçu l'inoculation de S. brunnea ont montré une diminution significative des concentrations de Cu, Pb et Sn dans le sol. L'inoculation avec R. irregularis ainsi que la double inoculation n'ont pas influencé de manière significative la production de biomasse et les niveaux d’éléments traces du sol. Le résultat de cette étude apporte des connaissances sur la diversité et l’écophysiologie des microbes de la rhizosphère associés aux plantes de croissance spontanée à la suite d’inoculations répétées. De plus ils montrent le potentiel de l’utilisation de champignons mycorhiziens afin d’améliorer la santé et croissance des plantes dans des milieux pollués et toxiques. Ils soulignent aussi l'importance de la sélection des plantes afin de faciliter leur gestion efficace et accélérer les processus de remise en état des terres. / The rhizosphere microbiome generally refers to the bacterial, archaea, and fungal communities and their genetic material that closely surrounds the root systems of plants. The metagenome of this microbiome has been called the second genome of the plant because it is able to take advantage of several functions that it lacks. It is a vibrant community of microorganisms, from which part of the species has developed long-lasting, specific and intimate interactions with plant roots. Recent advances in the study of plant-microbe interactions have demonstrated their considerable impact on plant growth, nutrition and health. The rhizosphere microbiota is complex with a dynamic spatio-temporal structure which adapts rapidly to biotic and abiotic stresses. Considering the importance of the rhizosphere microbiome to plant health, accurate information about their associated microbes is of utmost importance in deciphering the mechanisms of plant adaptation to microbiome-mediated stress, and understanding how plants recruit key microbial taxa to better cope with stressful conditions. To do this, we conducted three studies to shed light on the factors that play a role in the recruitment and structure of the microbiome of the rhizosphere of plants in stressed environments. First, we tested whether repeated inoculations with Proteobacteria influenced the productivity of plants and the microbial communities associated with the rhizosphere of four plant species growing in sediments contaminated with petroleum hydrocarbons. A mesocosm experiment was carried out in design of randomized blocks with two factors: 1) the presence or absence of four plant species collected in a sedimentation basin of a former petrochemical plant, and 2) inoculation or not with a bacterial consortium made up of ten isolates of Proteobacteria. The plants were grown in the greenhouse for four months. MiSeq amplicon sequencing, targeting the bacterial 16S rRNA gene and the fungal ITS, was used to assess the microbial community structures of sediments from planted and unplanted microcosms. Our results showed that while inoculation caused a significant change in microbial communities, the presence of the plant and its specific identity had a stronger influence on the structure of the microbiome in sediments contaminated with petroleum hydrocarbons. Next, using the same experimental setup, we used MiSeq amplicon sequencing targeting the 18S rRNA gene to assess AMF community structures in the roots and rhizosphere of plants growing in contaminated and uncontaminated substrates. We also studied the contribution of the specific identity of plants and the biotope (plant roots and rhizospheric soil) in the formation of associated AMF assemblages. Our results showed that while inoculation caused a significant change in AMF communities, substrate contamination had a much stronger influence on their structure, followed by biotope and plant identity to a lesser extent. In addition, inoculation dramatically increased plant biomass production and was associated with decreased dissipation of petroleum hydrocarbons in contaminated soil. The result of this study provides knowledge on the factors influencing the diversity and community structure of AMF associated with plants in stressed environments following repeated inoculations of a bacterial consortium. Finally, we tested the effect of inoculating trees with specific mycorrhizal fungi on their survival and growth, as well as the extraction of trace metals. To do this, a field experiment was carried out in which we cultivated the Salix miyabeana "SX67" clone on the site of a decommissioned industrial landfill and inoculated the shrubs with the arbuscular mycorrhizal fungus Rhizophagus irregularis, the ectomycorrhizal fungus Sphaerosporella brunnea, or a mixture of both. After two growing seasons, willows inoculated with the fungus S. brunnea produced a significantly higher biomass. Ba, Cd and Zn were found to accumulate in the aerial parts of plants, where Cd had the highest bioconcentration factor values in all treatments. In addition, the plots where the willows were inoculated with S. brunnea showed a significant decrease in the concentrations of Cu, Pb and Sn in the soil. The inoculation with R. irregularis as well as the double inoculation did not significantly influence the biomass production and the soil trace elements levels The result of this study provides insight into the diversity and ecophysiology of rhizosphere microbes associated with spontaneously growing plants following repeated inoculations. In addition, they show the potential of using mycorrhizal fungi to improve plant health and growth in polluted and toxic environments. They also stress the importance of plant selection to facilitate their efficient management, in order to speed up land reclamation processes.

Maize is the dominant crop in northwestern Germany and is mostly cultivated on sandy soils. Additionally, due to intensive livestock husbandry and biogas production, large amounts of liquid manures are produced. The current farm practice leads to high N and P surpluses at field level accompanied by environmental pollution, like nitrate leaching, eutrophication of non-agricultural ecosystems, and N2O emissions. The accruing liquid manures are often used for maize fertilization. Thereby, slurries are mainly broadcast applied using trailing hose applicators followed by incorporation into the topsoil. In addition, a mineral N P starter fertilizer (MSF) is band-applied below the seed-corn at planting to overcome the limited nutrient availability during the early growth stages. Using a slurry injection technique below the maize row before planting might serve a substitute for MSF. Addition of a nitrification inhibitor (NI) into the slurry before injection seems to be an option to further decrease N losses. The objectives of this thesis were to compare the current and novel fertilizing strategies with a special focus on soil mineral nitrogen (SMN) dynamics and plant P, zinc (Zn) and manganese (Mn) status. For both issues the effect of adding a NI into the slurry was investigated. To characterize the SMN dynamics after slurry injection an appropriate soil sampling strategy had to be developed. Therefore, three consecutive field trials were conducted. The first testing of the new soil sampling approach was implemented in an existing experiment where the slurry was injected at a depth of 12 cm (upper rim) below the soil surface. The soil profile (75 cm wide) centered below the maize row was sampled using a grid-like approach to a depth of 90 cm. Around the injection zone, soil monoliths (SM) were sampled using a purpose-built soil shovel. Below the SMs and in the interrow space (15 and 30 cm distance to the row) a standardized auger procedure was used. The second experiment aimed to improve the sampling strategy with focus on sample homogenization quality and necessary sample sizes per pooled sample. In the third experiment this improved sampling strategy was validated. Results from the first testing of the sampling procedure showed that the strategy is suitable, although some problems occurred. Especially the high spread in values among the replications caused high coefficients of variation (CV; mostly 40 – 60%). The improvement trial revealed that for the SM, which contains the slurry band, an intensive homogenization is required. In addition, suitable sample sizes (twelve auger samples and six soil monolith samples per pooled sample) have to be collected to obtain reliable SMN values. Following this enhanced sampling strategy in the final validation trial, the spread in values was considerably reduced and resulted in CV values of mostly < 20%. The method can be adapted to other fertilizer placement strategies and further row crops. To compare both fertilizing strategies with respect to the spatial and temporal SMN dynamics as well as to the plant nutrient status two field trials were conducted using pig slurry on sandy soils in 2014 and 2015. Four treatments were tested: unfertilized control, broadcast application + MSF, injection, and injection + NI. Soil samples were taken using the new sampling strategy at several dates during the growing season. Plant samples were simultaneously collected to evaluate the plant P, Zn, and Mn status at different growth stages. In 2014, all fertilized N was displaced from the top soil layer of the broadcast treatment until the 6-leaf stage due to heavy rainfall, while N displacement was significantly smaller after slurry injection. The lateral movement of injected slurry N was negligible. In 2015, almost no displacement of fertilized N out of the top soil layer occurred independently of treatments, due to distinctly lower rainfall. The release of slurry N was delayed following broadcast application and large SMN concentrations were detected in the injection zones until the 10-leaf stage. The addition of a NI resulted in significantly increased NH4-N shares in the injection zone throughout the early growth stages (+ 46% in 2014 and + 12% in 2015 at 6-leaf stage). Thus, in 2014 SMN displacement was delayed, and in 2015 increased SMN concentrations were found around the slurry band, most probably due to lower N losses via denitrification. Furthermore, NI addition significantly increased the nutrient uptake by maize during early growth in both years. With P deficiency due to cold weather conditions in 2015, broadcast application showed higher P uptake until the 6-leaf stage (36 – 58%), while it was lower at the 8- (32%) and 10- (19%) leaf stages compared to slurry injection (+ NI). Zn availability was enhanced during early growth after slurry injection (+ NI) and Zn as well as Mn uptake were higher at harvest. Furthermore, dry matter yields were higher (2014) or equal (2015) compared to broadcast application. The P balances were decreased by 10 – 14 kg P ha-1, while Zn and Mn balances were excessive independent of treatments. The field trials showed that after slurry injection, especially when combined with a NI, the applied nitrogen is located in a soil zone with better spatial availability for plant roots compared to broadcast application. Furthermore, the MSF can be substituted without affecting early growth of maize. In conclusion, slurry injection leads to equal (or even higher) yields and enables farmers in northwestern Germany to reduce the P and N surpluses. This would support several goals concerning sustainable land use: Lower pollution of ground and surface waters, reduced emission of NH3, more efficient use of the limited rock P reserves, and less need of transporting organic manures out of regions with intensive animal husbandry and/or biogas production. However, slurry injection enhances the risk of N2O emissions, which contributes to climate change. Thus, for a final evaluation of the environmental impact a life cycle assessment would be worthwhile.

No description available.