Low plant available phosphorus limits legume growth and biological nitrogen fixation (BNF). This study examined, under controlled conditions, the relationship between soil phosphorus and alfalfa and soybean BNF on two contrasting low-P soils (Ontario and Nova Scotia) from organic dairy farms. Soluble P was applied up to 135 mg P kg-1. An optimum range of 45 to 90 mg kg-1 applied P increased soybean plant growth, nodulation, N and P uptake and BNF. Significant effects of soil type reflected greater N supplying ability and lower P sorption for the Ontario soil. Alfalfa response to soluble P application was not as apparent. In addition three potentially organically acceptable amendments (MSW compost, Crystal Green® struvite and partially solubilized rock phosphate) were evaluated as alternate sources of plant available P. Compost and struvite, applied at moderate rates, sufficiently supplied P to increase plant growth and BNF comparably to that found for soluble P fertilizer.

No description available.

Symbiotic interaction of Rhizobium with roots of legume plants leads to the formation of root nodules that are capable of reducing atmospheric nitrogen. In nodule cells, bacteria are always enclosed inside the peribacteroid membrane (pbm) which is derived from the plant cell plasma membrane. The pbm mediates all the molecular exchanges between the host plant and bacteria. The pbm of soybean (Glycine max) nodules induced by Bradyrhizobium japonicum was characterized for the presence of nodule-specific proteins (nodulins), which may be involved in some of the nodule-specific roles the pbm plays. The pbm was found to contain a number of nodulins, and most of them were shown to be of plant origin. The peribacteroid fluid (pbf) was also purified and shown to contain nodulins. Some of the pbf proteins were highly glycosylated. Immunological localization at the electron microscopic level has shown that pbm nodulins are inserted specifically in the pbm; almost none are detected in the plasma membrane (pm) of infected cells even though the pbm is derived from it. The amino acid sequences of three different pbm nodulins (nodulin-23, -24 and -26) do not share significant primary structure homology, neither do their tentative secondary structures. Our results do not preclude targeting based on pbm-nodulin-specific feature(s) of the proteins; but it is possible that pbm-specific localization is achieved by differential rates of membrane synthesis, or turnover, in pbm as compared to the plasma membrane.

Soybean [Glycine max (L.) Merr.] is a tropical to subtropical legume that requires root zone temperatures (RZTs) in the 25 to 30°C range for optimal symbiotic activity. The inability of soybean to adapt to cool soil conditions limits its development and yield in short season areas. In particular, nodulation and nitrogen fixation by this crop species is sensitive to cool RZTs. The isoflavone genistein, which is the most effective plant-to-bacterium signal in the soybean nitrogen fixation symbiosis, has been used to pretreat Bradyrhizobium japonicum inocula. This resulted in increased soybean nodulation and nitrogen fixation in several studies, indicating that genistein preincubated inocula could overcome low RZT inhibition of plant growth and yield. The effectiveness of isoflavones was found to vary among soybean cultivars. Some legume cultivars apparently supply limiting amounts of the flavonoids. The objective of this thesis was to determine how soybean cultivars of different maturity groups would respond to genistein incubation of B. japonicum prior to inoculation. Two field experiments were conducted in 1997 and 1998 involving eleven soybean cultivars of three maturity groups organized in a randomized complete block design. Cells of B. japonicum, treated with genistein or not, were applied onto seeds in the furrow at the time of planting. The results of this study indicated that genistein application increased nodule number and nodule dry matter per plant, increased plant nitrogen content, grain protein and grain yield of all cultivars. There was no interaction between maturity group and genistein application, and there was no correlation between maturity groups and increase in nodulation, total biomass, nitrogen content and yield due to genistein treatment. Thus, responses of soybean cultivars of different maturity groups to genistein treatment are not related to maturity and/or yield potential.

This project was designed to assess the ability of natural sediment microbial communities and single species microbial populations to preferentially utilize inorganic forms of nitrogen (ammonium, NH4-N, and nitrate, NO3-N, specifically). The first chapter addressed two primary questions: 1) Do sediment microbial communities preferentially assimilate NH4-N or NO3-N?; and, 2) Does preferential uptake of nitrogen change with increased NH4-N or NO3-N availability? The second chapter furthered these analyses by assessing shifts in microbial nitrogen assimilation in response to sustained nitrogen enrichments. Primary questions addressed were: 1) Are microbial communities able to adapt to nitrogen enrichment and preferentially utilize a more available source?; and, 2) Are initial microbial responses to nitrogen enrichment different from sustained responses? Questions were addressed with in vitro laboratory experiments quantifying microbial activity. Overall, microbial community activity changed in response to the form of nitrogen available, enrichment type, and duration of exposure. Data demonstrate sediment microbial communities in the Midwestern US may prefer NO3-N over other forms of nitrogen. However, microbial communities became saturated with NO3-N with increases in concentrations >0.75 mg NO3-N/L. Microbial communities were able to adapt to higher nitrogen concentration and increase rates of assimilation for both NH4-N and NO3-N. Thus, microbial communities are robust in response to nitrogen increases in and ecosystem, even in high nitrogen environments like the Midwestern US. / Preferential uptake of available nitrogen forms -- Adaptive uptake in microbial communities. / Department of Biology

No description available.

Pseudomonas saccharophila could fix N$ sb2$ under micro-aerobic conditions, heterotrophically and chemolithotrophically. Uptake hydrogenase activity under heterotrophic conditions had no effect on the O$ sb2$ sensitivity of nitrogenase. H$ sb2$ induced whereas sucrose and O$ sb2$ repressed hydrogenase synthesis. Sucrose and O$ sb2$ did not inhibit hydrogenase activity. Hydrogenase and urease were located in the membrane and soluble fractions, respectively. Nickel stimulated growth, hydrogenase expression, and nitrogenase activity under N-limited chemolithotrophic conditions. Hydrogenase synthesis specifically required nickel and its repression by O$ sb2$ was alleviated by increasing the nickel concentration. Incorporated $ sp{63}$Ni$ sp{2+}$ was about 3 times higher in the soluble than in the membrane fraction. The short-term uptake of nickel was energy-independent and had an apparent $K sb{m}$ of 31.7 uM and $V sb{max}$ of 3.8 nmol Ni$ sp{2+}$ (mg protein)$ sp{-1}$min$ sp{-1}$. / A counting method for heterotrophic and chemolithotrophic N$ sb2$-fixing H$ sb2$-oxidizing bacteria was developed. The white bean rhizosphere soil showed relatively high numbers of these bacteria.

Graduation date: 1987

The transcriptional promoter sequences for the Rhizobium meliloti nitrogen fixation genes nifA and nifB were cloned to a β-galactosidase gene fusion plasmid vector and transferred by homologous recombination to a specialized transducing phage. The promoter fusions were then transduced to a recombination deficient strain of Escherichia coli as single-copy lysogens and analysed under defined aerobic and anaerobic conditions. The lysogenic strains contained plasmids encoding either of two transcriptional activator proteins, NifA or FixJ, produced from a constitutive plasmid promoter. The expression of the nifA and the nifB promoters was found to be sensitively regulated by the carbon source used for anaerobic fermentation or anaerobic respiration, the redox potential of the terminal electron acceptor used for anaerobic respiration, and the growth phase of anaerobic cultures. The repression of nit promoter expression by oxygen respiration was specifically compared to anaerobic respiration of alternative electron acceptors. Both nifA and nifB promoter expression decreased exponentially as the reduction potential of the terminal respiration reaction increased. The repressive effect of oxygen appears to be due soley to the exponential relationship between nit promoter expression and the redox potential of oxygen respiration. In addition to separate fusions of the nifA and nifB promoters to β-galactosidase, a single-copy fusion of the entire nifA-nifB region was constructed. In this construct, plasmid-encoded FixJ protein stimulated the expression of a chromosomal nifA gene to produce the NifA protein, which then stimulated the expression of the nitB promoter. This strain produced 20-fold lower activity than a strain in which nifB promoter expression was stimulated by plasmid-encoded NifA protein. Finally, the nifA locus was found to contain a transcriptionally active element, oriented opposite to the nifA promoter. / Graduation date: 1992

No description available.