The environmental conditions driving the microbial community dynamics in crop soils remain unclear. Here, we focused on the spatial and temporal dynamics of microbial communities in soils cultivated with sugarcane under different soil managements, during two years. Our work was divided into three essential parts, where i) we discuss ecological models and theories for the microbial exploration in crop soils, arguing that those ecological models, which partitioned the microbial communities, may increase the resolution of the environmental and the microbial interactions; ii) we developed a probabilistic model based on the occurrence frequency of microorganisms across systems identifying the core microbial community. The model is based on the Poisson distribution, and it was tested in four datasets available in the Earth Microbiome Project; iii) we identified the core bacterial and fungal communities across soils cultivated with sugarcane, verifying which abiotic components could drive the composition of groups. We increased the resolution of the environmental and the microbial interactions, showing that the core and the variable microbial communities are driven by distinct abiotic components. We also observed that the core and variable microbial communities harbor distinct potential functionality, as nitrogen fixation being more predicted to the core bacterial commmunity, and nitrification process for the variable bacterial community. Our finds increase the knowledge of microbial dynamics and functionality, helping to reveal and explore the crop system microbiome. / As condições ambientais que podem modular a dinâmica da comunidade microbiana em solos de culturas são pouco conhecidas. O presente trabalho foi dividido em três partes essenciais, onde i) discutiu-se modelos e teorias ecológicas para a exploração microbiana em solo agrícolas, argumentando-se que os modelos ecológicos que particionam as comunidades microbianas, poderiam aumentar a resolução entre interações microbianas e o ambiente, ii) desenvolveu-se um modelo probabilístico baseado na freqüência de ocorrência de microorganismos através de sistema identificando a comunidade microbiana \"core\". O modelo baseou-se na distribuição de Poisson, sendo este testado em quatro conjuntos de dados disponíveis no Projeto \"Earth Microbiome\", e iii) identificou-se as comunidades bacterianas e fúngicas core em solos cultivados com cana-de-açúcar, verificando-se quais componentes abióticos poderiam modular a composição dos grupos. Com isso, elevou-se a resolução das interações ambiental e microbiana, indicando que o core microbiano e as comunidades microbianas variáveis são moduladas por componentes abióticos distintos. Observou-se também que as comunidades core e variável possuem funcionalidade potencial distinta, como fixação de nitrogênio mais predita para o core bacteriano e processo de nitrificação para a comunidade variável de bactérias. Os resultados do presente trabalho elevam o conhecimento da dinâmica e funcionalidade microbiana, ajudando a revelar e explorar o microbioma do sistema de cultivo.

The environmental conditions driving the microbial community dynamics in crop soils remain unclear. Here, we focused on the spatial and temporal dynamics of microbial communities in soils cultivated with sugarcane under different soil managements, during two years. Our work was divided into three essential parts, where i) we discuss ecological models and theories for the microbial exploration in crop soils, arguing that those ecological models, which partitioned the microbial communities, may increase the resolution of the environmental and the microbial interactions; ii) we developed a probabilistic model based on the occurrence frequency of microorganisms across systems identifying the core microbial community. The model is based on the Poisson distribution, and it was tested in four datasets available in the Earth Microbiome Project; iii) we identified the core bacterial and fungal communities across soils cultivated with sugarcane, verifying which abiotic components could drive the composition of groups. We increased the resolution of the environmental and the microbial interactions, showing that the core and the variable microbial communities are driven by distinct abiotic components. We also observed that the core and variable microbial communities harbor distinct potential functionality, as nitrogen fixation being more predicted to the core bacterial commmunity, and nitrification process for the variable bacterial community. Our finds increase the knowledge of microbial dynamics and functionality, helping to reveal and explore the crop system microbiome. / As condições ambientais que podem modular a dinâmica da comunidade microbiana em solos de culturas são pouco conhecidas. O presente trabalho foi dividido em três partes essenciais, onde i) discutiu-se modelos e teorias ecológicas para a exploração microbiana em solo agrícolas, argumentando-se que os modelos ecológicos que particionam as comunidades microbianas, poderiam aumentar a resolução entre interações microbianas e o ambiente, ii) desenvolveu-se um modelo probabilístico baseado na freqüência de ocorrência de microorganismos através de sistema identificando a comunidade microbiana \"core\". O modelo baseou-se na distribuição de Poisson, sendo este testado em quatro conjuntos de dados disponíveis no Projeto \"Earth Microbiome\", e iii) identificou-se as comunidades bacterianas e fúngicas core em solos cultivados com cana-de-açúcar, verificando-se quais componentes abióticos poderiam modular a composição dos grupos. Com isso, elevou-se a resolução das interações ambiental e microbiana, indicando que o core microbiano e as comunidades microbianas variáveis são moduladas por componentes abióticos distintos. Observou-se também que as comunidades core e variável possuem funcionalidade potencial distinta, como fixação de nitrogênio mais predita para o core bacteriano e processo de nitrificação para a comunidade variável de bactérias. Os resultados do presente trabalho elevam o conhecimento da dinâmica e funcionalidade microbiana, ajudando a revelar e explorar o microbioma do sistema de cultivo.

The Upper Mississippi River (UMR) basin contributes over 50,000 metric tons of nitrogen (N) to the Gulf of Mexico each year, resulting in a “dead zone” inhospitable to aquatic life. Land-applied N (fertilizer) in the corn-belt is attributed with a majority of the N-load reaching the Gulf and is difficult to treat as run-off is considered a non-point source of pollution (i.e. not from a pipe). One solution to this “grand challenge” of intercepting N pollution is utilizing filter-feeding organisms native to the UMR. Freshwater mussel (order Unionidae) assemblages collectively filter over 14 billion gallons of water, remove tons of biomass from overlying water, and sequester tons of N each day. Our previous research showed mussel excretions increased the sediment porewater concentrations of ammonium by 160%, and indirectly increased nitrate and nitrite by 40%, presumably from microbial degradation of ammonium. In response, the goal of this research was to characterize how mussels influenced microbial communities (microbiome) to determine the fate of N in UMR sediment. First, we used qPCR and non-targeted amplicon sequencing within sediment layers to identify the N-cycling microbiome and characterized microbial community changes attributable to freshwater mussels. qPCR identified that anaerobic ammonium oxidizing (anammox) bacteria were increased by a factor of 2.2 at 3 cm below the water-sediment interface when mussels were present. Amplicon sequencing of sediment at depths relevant to mussel burrowing (3 and 5 cm) showed that mussel presence reduced microbial species richness and diversity and indicated that sediment below mussels harbored distinct microbial communities. Furthermore, mussels increased the abundance of ammonia oxidizing bacteria (family Nitrosomonadaceae), nitrite oxidizing bacteria (genus Nitrospira), but decreased the abundance of ammonia oxidizing archaea (genus Candidatus Nitrososphaera), and microorganisms which couple denitrification with methane oxidation. These findings suggested that mussels may enhance microbial niches at the interface of oxic and anoxic conditions, presumably through excretion of N and burrowing activity. In response, we performed metagenomic shotgun sequencing to identify which genes of the microbiome were most impacted by mussels. We hypothesized that genes responsible for ammonia and nitrite oxidation would be greater in the sediment with mussel assemblages. We found the largest abundance of N-cycling genes were responsible for nitrate reduction and nitrite oxidation, which is corroborated by the high concentration of nitrates in UMR water. Linear discriminant analysis statistical analyses showed nitrification genes were most impacted by mussels, and this presented an opposing effect on genes responsible for producing nitrous oxide, a potent greenhouse gas. Further investigation showed an increased abundance of a novel organism capable of completely oxidizing ammonia to nitrate (Candidatus Nitrospira inopinata) and coexisted with metabolically flexible Nitrospira (sp. moscoviensis), likely enhancing both carbon and N-cycling. We demonstrated that native mussels harbor a unique niche for N-cycling microorganisms with large metabolic potentials to degrade mussel excretion products. Our findings suggest the ecosystem services of mussels extend beyond water filtration, and includes enhanced biogeochemical cycling of carbon, N, and reduces the potential for a potent microbially-produced greenhouse gas. Ultimately, this research could be used to advocate for mussel habitat restoration in the UMR to lessen the impacts of non-point pollution.

Coral reefs are biologically and economically important ecosystems underpinned by corals that are able to flourish in oligotrophic waters due to their mutualistic association with dinoflagellate symbionts (genus Symbiodinium). Symbiodinium are strictly intracellular, residing within the gastrodermal tissues of the coral host, and contributing the majority of the coral’s energy requirements. Coral reefs are in rapid decline due to a range of threats such as local human influences, bleaching (loss of Symbiodnium and/or reduction of pigment), disease and ocean acidification, to which links to climate change have been made. The close association of corals and a diverse community of microbes led to development of the coral holobiont hypothesis, in which a range of microorganisms (e.g Bacteria) form a functionally-relevant mutualistic relationship with corals and Symbiodinium. This thesis aimed to fill knowledge gaps in the coral holobiont hypothesis and the host-microbe interactions within this system, including pathogen interactions and coral immune system functioning. This thesis revealed that host-microbe interactions in corals are complex, and that the underlying mechanisms of immunity and symbiosis may be similar. The findings corroborate the idea that corals maintain specific bacterial communities that have potential probiotic and nutritional value. In particular, a group of common coral associates were identified, and it is suggested that members of this group are globally occurring key associates. Corals affected by a disease previously described as “White Syndrome” were observed to undergo pronounced changes in their microbial community structure in comparison to healthy colonies. However, in contrast to previous findings, no single pathogen could be identified as the causative agent of the disease syndrome, and it is speculated that corals experiencing altered health status result in a breakdown of the resident associated microbial community structure. Culturable bacterial isolates from corals were shown to affect the growth of each other and in particular some species had great inhibitory properties. Hence, the presence of some bacterial species has the potential to influence the all over structure of the coral associated microbial community. It was also shown that changed environmental conditions may alter the growth conditions for coral associated bacteria in mucus. It is suggested that increased replication is needed in studies of bacterial assemblages on corals, as variability between coral species and sites were observed. In addition, studies of the role of coral microbial communities in health and disease should broaden their focus to more thoroughly consider the role of the coral holobiont, especially with regards to the coral host. This thesis identified the first functional Pattern Recognition Protein (PRP), a C-type lectin named Millectin, in scleractinian corals. Millectin was isolated by affinity chromatography and was shown to bind to bacterial pathogens as well as coral Symbiodinium symbionts. Gene expression of Millectin was upregulated in response to immune stimuli and the lectin was further abundantly expressed in the tissues of corals, suggesting a major role for this protein in system functioning and immunity. Further research into Millectin and a complement factor C3 homolog suggested that these molecules may have been co-opted into the equally important role of symbiont recruitment. Gene expression analysis of C3 also indicated this molecule may be involved in responses to tissue trauma. Millectin shows variability in the binding region, and hence, is the earliest evolutionary representative to date of a variable PRP. This finding, and the observed ancestral relation with vertebrate homologs, provided further information on the evolution of the innate immune system and gives further insight into invertebrate immunity.

Thesis (Ph.D. (Environmental Science)--North-West University, Potchefstroom Campus, 2007.

Adoption of no-till (NT) and reduced tillage management is widespread on the Canadian prairies and together form the basic platform of soil management upon which most crop production is based. Elimination of tillage in cropping systems changes the physical and chemical characteristics of the soil profile and can affect crop growth and ultimately yield. As such, understanding how soil biota, as drivers of nutrient turnover, adapt to NT is important for maximizing crop productivity and mitigating environmental damage in agroecosystems. This work aims to achieve a greater understanding of microbial community structure and function in long-term NT versus conventionally tilled (CT) soils. Community phospholipid and DNA fingerprinting did not reveal any consistent tillage-induced shifts in microbial community structure, but demonstrated a clear influence of depth within the soil profile. While tillage did not result in broad changes in the community structure, total, bacterial and fungal biomass was consistently greater near the surface of NT soils. Further examination at one site near Swift Current, SK revealed differences in microbial biomass and community structure in NT and CT in field-formed aggregate size fractions. Measurement of mineralization and nitrification at the same site indicated that differences in the early-season turnover of N may be related to physical rather than microbial differences in NT and CT soils. Potential nitrification was higher prior to seeding than mid-season, was not affected by tillage and was correlated with ammonia oxidizer population size of archaea, but not bacteria. This work indicates that edaphic soil properties and spatial distribution of resources in the soil profile, rather than tillage management, are the primary factors driving microbial community structure in these soils.

Phytoremediation systems for petroleum hydrocarbons rely on a synergistic relationship between plants and their root-associated microbial communities. Plants exude organic compounds through their roots, which increase the density, diversity and activity of plant-associated microorganisms, which in turn degrade hydrocarbons. Understanding the mechanisms driving this relationship poses one of the more intriguing challenges in phytoremediation research. This study was designed to address that challenge. Plant-microbe interactions in a weathered-hydrocarbon contaminated soil were examined under controlled growth chamber, and field conditions. In both environments single-species grass treatments initially facilitated greater total petroleum hydrocarbon (TPH) degradation than <i> Medicago sativa </i> (alfalfa), mixed species, or control treatments. In growth chamber studies increased degradation was linked to increased aliphatic-hydrocarbon degrader populations within the rhizosphere. Under field conditions, specific recruitment of endophytic aliphatic-hydrocarbon degraders in response to high TPH levels may have facilitated increased degradation by the grass <i> Elymus angustus</i>(Altai wild rye, AWR). AWR stably maintained these communities during times of local drought, enabling them to act as subsequent source populations for rhizosphere communities. The broad phylogenetic diversity of AWR endophytes, compared to the <i> Pseudomonas</i>-dominated communities of other plants, contributed to the observed stability. The relative composition of exudates released by plants also impacted both degradation activity and potential. Alfalfa released higher concentrations of malonate, which hindered degradation by decreasing metabolic activity and concomitantly inhibiting catabolic plasmid transfer. In contrast, AWR exudates contained high levels of succinate, which was linked to increased catabolic gene expression and plasmid transfer. A reciprocal relationship between exudation patterns and endophytic community structure likely exists, and both parameters have a specific influence on rhizosphere degradation capacity. In this study, grasses were more successful in maintaining the specific balance of all parameters required for the transfer, preservation, and stimulation of hydrocarbon catabolic competency.

A biofilter removes biodegradable contaminants from air by passing it through a biologically-active packed bed. The microorganism community is of fundamental interest but has been the focus of few studies. This work is an investigation of the bacterial community based on the potential functional diversity of the community. <br /><br /> A number of experiments were performed in laboratory-scale biofilters using ethanol as a model contaminant. All biofilters were able to remove the ethanol with elimination capacities in the range 80 to 200gVOCm^-3h^-1; these values are comparable with published literature. Natural organic media (peat or compost) was used as packing. <br /><br /> The potential functional diversity of the community was assessed by Community-Level Physiological Profiling (CLPP) using sole-Carbon Source Utilisation Profile (CSUP). Community samples were used to inoculate Biolog EcoPlates^TM: microplates containing a selection of 31 different carbon-substrates and an indicator dye responding to bacterial growth. This technique was found to be sensitive to changes in the community structure over time and location. <br /><br /> Results showed that the community in samples taken close together (over a scale of a few centimetres) are similar and that relatively small media samples (0. 5 to 1 g) provide reproducible information. A study of a single biofilter indicated stratification of the community occurring with the community near the inlet diverging from that near the middle and outlet of the unit; this is attributed to the ethanol being degraded in the upper part of the column and the lower part of the column not being subjected to ethanol loading. In a study of two units at a higher loading rate, stratification was not observed over a period of weeks; it is suggested that the stratification may develop over this timescale as a result of the presence or absence of the Volatile Organic Compound (VOC) and not due to differences in concentration. <br /><br /> An acclimation period of 7 to 10 days was observed before near-complete removal of ethanol was attained. Monitoring of the community suggested a subsequent shift in diversity. It is suggested that the initial acclimation period is due to biofilm formation and the subsequent shift in community diversity is due to re-organisation of the community as species specialise. In a portion of the biofilter with minimal ethanol exposure, a sudden shift in community is observed after a period of some weeks. This may reflect changes as a result of starvation and indicates that periods of shut-down (when the biofilter is not loaded) may affect the community. <br /><br /> Two studies of biofilters operating in parallel were carried out. The first provided evidence of a divergence in the communities over a period of two weeks. In the second, communities in the two units underwent changes over time but observations from both units at any one time were similar. This demonstrates that biofilters set-up and operated in a similar manner may maintain similar communities but that this is not necessarily the case. This has implications for the reproducibility of laboratory experiments and for the variation of community structure with horizontal position in industrial units.

A biofilter removes biodegradable contaminants from air by passing it through a biologically-active packed bed. The microorganism community is of fundamental interest but has been the focus of few studies. This work is an investigation of the bacterial community based on the potential functional diversity of the community. <br /><br /> A number of experiments were performed in laboratory-scale biofilters using ethanol as a model contaminant. All biofilters were able to remove the ethanol with elimination capacities in the range 80 to 200gVOCm^-3h^-1; these values are comparable with published literature. Natural organic media (peat or compost) was used as packing. <br /><br /> The potential functional diversity of the community was assessed by Community-Level Physiological Profiling (CLPP) using sole-Carbon Source Utilisation Profile (CSUP). Community samples were used to inoculate Biolog EcoPlates^TM: microplates containing a selection of 31 different carbon-substrates and an indicator dye responding to bacterial growth. This technique was found to be sensitive to changes in the community structure over time and location. <br /><br /> Results showed that the community in samples taken close together (over a scale of a few centimetres) are similar and that relatively small media samples (0. 5 to 1 g) provide reproducible information. A study of a single biofilter indicated stratification of the community occurring with the community near the inlet diverging from that near the middle and outlet of the unit; this is attributed to the ethanol being degraded in the upper part of the column and the lower part of the column not being subjected to ethanol loading. In a study of two units at a higher loading rate, stratification was not observed over a period of weeks; it is suggested that the stratification may develop over this timescale as a result of the presence or absence of the Volatile Organic Compound (VOC) and not due to differences in concentration. <br /><br /> An acclimation period of 7 to 10 days was observed before near-complete removal of ethanol was attained. Monitoring of the community suggested a subsequent shift in diversity. It is suggested that the initial acclimation period is due to biofilm formation and the subsequent shift in community diversity is due to re-organisation of the community as species specialise. In a portion of the biofilter with minimal ethanol exposure, a sudden shift in community is observed after a period of some weeks. This may reflect changes as a result of starvation and indicates that periods of shut-down (when the biofilter is not loaded) may affect the community. <br /><br /> Two studies of biofilters operating in parallel were carried out. The first provided evidence of a divergence in the communities over a period of two weeks. In the second, communities in the two units underwent changes over time but observations from both units at any one time were similar. This demonstrates that biofilters set-up and operated in a similar manner may maintain similar communities but that this is not necessarily the case. This has implications for the reproducibility of laboratory experiments and for the variation of community structure with horizontal position in industrial units.

Phytoremediation systems for petroleum hydrocarbons rely on a synergistic relationship between plants and their root-associated microbial communities. Plants exude organic compounds through their roots, which increase the density, diversity and activity of plant-associated microorganisms, which in turn degrade hydrocarbons. Understanding the mechanisms driving this relationship poses one of the more intriguing challenges in phytoremediation research. This study was designed to address that challenge. Plant-microbe interactions in a weathered-hydrocarbon contaminated soil were examined under controlled growth chamber, and field conditions. In both environments single-species grass treatments initially facilitated greater total petroleum hydrocarbon (TPH) degradation than <i> Medicago sativa </i> (alfalfa), mixed species, or control treatments. In growth chamber studies increased degradation was linked to increased aliphatic-hydrocarbon degrader populations within the rhizosphere. Under field conditions, specific recruitment of endophytic aliphatic-hydrocarbon degraders in response to high TPH levels may have facilitated increased degradation by the grass <i> Elymus angustus</i>(Altai wild rye, AWR). AWR stably maintained these communities during times of local drought, enabling them to act as subsequent source populations for rhizosphere communities. The broad phylogenetic diversity of AWR endophytes, compared to the <i> Pseudomonas</i>-dominated communities of other plants, contributed to the observed stability. The relative composition of exudates released by plants also impacted both degradation activity and potential. Alfalfa released higher concentrations of malonate, which hindered degradation by decreasing metabolic activity and concomitantly inhibiting catabolic plasmid transfer. In contrast, AWR exudates contained high levels of succinate, which was linked to increased catabolic gene expression and plasmid transfer. A reciprocal relationship between exudation patterns and endophytic community structure likely exists, and both parameters have a specific influence on rhizosphere degradation capacity. In this study, grasses were more successful in maintaining the specific balance of all parameters required for the transfer, preservation, and stimulation of hydrocarbon catabolic competency.