Seeing the forest for the trees: Tree species effects on soil microbial communities and nutrient cycling dynamics

Tree species influence soils above and belowground communities through leaf litter and root inputs. Soil microbial communities can directly influence tree growth and development through processes such as decomposition of leaves, and indirectly through chemical transformation of nutrients in soils as an influence of altered C:N ratios due to leaf litter and root inputs. This thesis aims to document some of the mechanisms by which trees influence soil microbial communities and nitrogen cycling processes like gross and net ammonification and nitrification. This thesis also aims to determine the role of site nitrogen status in modulating those tree species effects. The effects of tree species on ammonification and nitrification rates in forest floors and mineral soils were explored, and related to functional genetic markers for ammonia-oxidation by archaea and bacteria (amoA AOA and AOB), bacterial denitrification (nirS and nirK), and the general markers for bacteria (16S) and fungi (ITS). Two paired high-resolution laboratory methods were used to investigate the relationships between trees, soils, and the microbial communities, including molecular techniques such as quantitative polymerase chain reaction (qPCR) to target gene abundances in soils, and 15N pool-dilution experiments to understand how ammonium and nitrate are produced and consumed in soils. Soil samples were collected from two common garden experiments, named EP571 in Canada (Ribbons et al. 2016), and in Denmark, and both 15N and qPCR-based techniques were used to determine tree species effects and attribute N cycling processes to the abundances of functional genes. At EP571, western red cedar (Thuja plicata) forest floor nitrogen transformation rates differed from Douglas-fir (Pseudotsuga menziesii), Sitka spruce (Picea sitchensis), and western hemlock (Tsuga heterophylla), which corresponded with western red cedar having highest abundances of bacterial 16S and amoA AOA genes. A manipulative mesocosm (the Rhizotron) in Wales was used to determine how mixtures and monocultures of seedling species influenced tree growth, soil physical properties and soil microbial community structure and function within the first three years of growth. Within the Rhizotron experiment both alder (Alnus rubra) and Douglas-fir grew taller when grown in mixtures compared with monocultures of each species. Sycamore maple (Acer pseudoplatanus) acquired slightly greater aboveground biomass when planted in mixtures with common oak (Quercus robur), which attained less biomass in mixtures than monocultures. C:N ratios of leaves, stems, roots, and rhizospheric soils were determined to see if mixtures influenced C:N in trees. Rhizospheric soil microbial communities (including bacterial and fungal markers and the 4 genes tied to N cycling) were compared among the 4 tree species in the Rhizotron. Soil samples for microbial analyses were collected before seedlings were planted, and just before the experiment was harvested. These data show differences in height, biomass and C:N ratios between species can be observed at a seedling growth stage, but microbial communities may require longer exposure to develop. Lastly, the Bangor Diverse experiment was used to further explore diversity and mixture effects on soil microbial communities and N transformations. We found few mixture or monoculture tree species effects on mineral soil microbial communities or net nitrification or ammonification rates. Collectively, these stories shed light on the important functional role of soil microbes in forest soil N cycling. This thesis also highlights the use of isotope and microbial techniques for parsing out relationships between site, tree species identity and ecosystem functions, with the largest links observed between gross ammonification and microbial communities

Ant-mediated seed dispersal in a warmed world

Climate change affects communities both directly and indirectly via changes in interspecific interactions. One such interaction that may be altered under climate change is the ant-plant seed dispersal mutualism common in deciduous forests of eastern North America. As climatic warming alters the abundance and activity levels of ants, the potential exists for shifts in rates of ant-mediated seed dispersal. We used an experimental temperature manipulation at two sites in the eastern US (Harvard Forest in Massachusetts and Duke Forest in North Carolina) to examine the potential impacts of climatic warming on overall rates of seed dispersal (using Asarum canadense seeds) as well as species-specific rates of seed dispersal at the Duke Forest site. We also examined the relationship between ant critical thermal maxima (CTmax) and the mean seed removal temperature for each ant species. We found that seed removal rates did not change as a result of experimental warming at either study site, nor were there any changes in species-specific rates of seed dispersal. There was, however, a positive relationship between CTmax and mean seed removal temperature, whereby species with higher CTmax removed more seeds at hotter temperatures. The temperature at which seeds were removed was influenced by experimental warming as well as diurnal and day-to-day fluctuations in temperature. Taken together, our results suggest that while temperature may play a role in regulating seed removal by ants, ant plant seed-dispersal mutualisms may be more robust to climate change than currently assumed

Measures that can be used to teach critical thinking skills in nurse prescribers

Critical thinking is a pervasive skill that involves scrutinizing, differentiating, and appraising information and reflecting on the information gained in order to make judgements and inform clinical decisions. Studies inform us of the need for agreement on the approaches used to teach and measure critical thinking. Nurse prescribers undertake an advanced role that encompass the need to be able to make clinically based decisions about the appropriateness of a specific medication. This requires critical thinking attributes. A variety of teaching and learning approaches are offered which can be used by nurse educators to develop critical thinking skills in nurse prescribers

Bacterial metabolism of side chain fluorinated aromatics: cometabolism of 4-trifluoromethyl(TFM)-benzoate by 4-isopropylbenzoate grown Pseudomonas putida JT strains

Enzymes of the p-cymene pathway in Pseudomonas putida strains cometabolized the intermediate analogue 4-trifluoromethyl(TFM)benzoate. Three products, 4-TFM-2,3-dihydro-2,3-dihydroxybenzoate, 4-TFM-2,3-dihydroxy-benzoate and 2-hydroxy-6-oxo-7,7,7-trifluorohepta-2,4-dienoate (7-TFHOD) were identified chemically and by spectroscopic proterties.Certain TFM-substituted analogue metabolites of the p-cymene pathway were transformed at drastically reduced rates. Hammett type analysis of ring cleavage reactions of 4-substituted 2,3-dihydroxybenzoates revealed the negative inductive and especially mesomeric effect of substituents to be rate determining. Whereas decarboxylation of 3-carboxy-7-TFHOD was not affected by fluorine substitution the subsequent hydrolysis of 7-TFHOD proceeded very slowly. The negative inductive effect of the TFM-group probably inhibited heterolysis of the carbon bond between C5 and C6 of 7-TFHOD

Interactions of substrate and non-substrate effectors with p-hydroxybenzoate hydroxylase from

3,4-Dihydroxybenzoate (3,4-DOHB), 2,4-dihydroxybenzoate (2,4-DOHB), and benzoate facilitate the interaction of p-hydroxybenzoate hydroxylase with TPNH. The two dihydroxybenzoate effectors form 1:1 complexes with the enzyme, inducing large spectral perturbations and fluorescence quenching. The dissociation constants for 2,4-DOHB and 3,4-DOHB are 0.15 and 0.50 mM respectively. During the reaction of enzyme with TPNH and oxygen, all the 2,4-DOHB, <5% of the benzoate, and none of the 3,4-DOHB is hydroxylated.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/33509/1/0000006.pd