A potential biological role for microcystin in photosynthesis in Microcystis Aeruginosa
- Authors: Phelan, Richard Reginald
- Date: 2009
- Subjects: Microcystis aeruginosa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10320 , http://hdl.handle.net/10948/1285 , Microcystis aeruginosa
- Description: Neither the ecological role nor the metabolic function of microcystin is known. Cellular microcystin concentrations correlate to cellular nitrogen status for a given environmental phosphorous concentration and specific growth rate. Microcystin production is enhanced when the rate of nitrogen accumulation exceeds the relative specific growth rate and/or when cellular N:C ratios exceed the Redfield ratio as a function of reduced carbon fixation, suggesting enhanced production of microcystin under carbon stress. Additionally, a strong correlation between medium phosphate and carbon fixation, and the negative correlation between medium phosphate and microcystin combined with the cellular localization of microcystin in thylakoids supports a possible role for microcystin in enhancement of photosynthesis. Batch cultures of both Microcystis aeruginosa PCC7806 and a mcyA- knockout mutant of PCC7806 were therefore cultured at different light intensities and media treatments, so as to vary cellular N:C ratios and concentrations, and sampled for analysis of microcystin concentration, cell numbers and residual medium nitrates. Inter-strain differences in photosynthetic electron transfer rates and levels were monitored using a Hansatech PEA fluorometer and compared to cellular microcystin concentrations. An enhanced survival was observed at high light, where the toxic strain survived while the nontoxic strain became chlorotic. A strong correlation (r2 = 0.907, p< 0.001, N=22) between microcystin concentration and growth rate was observed at high light conditions. No such advantage was observed at optimal or low-light conditions and media composition had no significant effect on the relationship between toxicity and survival at high light. PCC7806 showed elevated PI(abs) values compared to the mcyA knockout strain, which indicates an increased stability of PSII. A strong correlation between PI(abs) and microcystin (r = 0.88, p< 0.005, N=15) was observed for cultures grown in modified BG11 containing 25 mM under continuous illumination of 37 μmol of photons m-2.s-1. No correlation was observed between PI(abs) and microcystin for the other treatments. The toxin producer had significantly higher values for density of active reaction centers and ii quantum efficiency compared to the mutant. A decrease in F0 in the mutant suggests degradation of the phycobiliproteins, whereas PCC7806 didn’t show a significant decrease in F0 Data indicate that microcystins play a role in photosynthesis by preventing chlorosis in saturating light conditions either by enhancing the redox stability of the phycobiliproteins or PS II, thus preventing photooxidation.
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- Date Issued: 2009
The immobilization of Microcystis aeruginosa PCC7806 on a membrane nutrient-gradostat bioreacator for the production of the secondary metobolites
- Authors: Strong, Peter James
- Date: 2002
- Subjects: Microcystis aeruginosa , Myrocystins , Bioreactors
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:11083 , http://hdl.handle.net/10948/283 , Microcystis aeruginosa , Myrocystins , Bioreactors
- Description: A module and an inoculation technique were developed that would allow for the efficient immobilization of Microcystis aeruginosa PCC7806 on a synthetic membrane. A variety of module types, membranes (ceramic, tubular polyethersulfone and externally skinless polyethersulfone capillary membrane), and methods of immobilization (adsorption, pressure filtration and a developed technique that involved drying a cell slurry on a membrane) were assessed. The morphological properties that affected the immobilization of Microcystis aeruginosa PCC7806, as well as the effects of immobilization upon cell morphology were assessed. Cells in the stationary growth phase, which had a well-developed extra-cellular polysaccharide layer and no gas vesicles, were optimal for immobilization. Microcystin production under immobilized conditions was assessed under different nitrate concentrations, light intensities, biofilm thickness and immobilization times. Additional work included assaying for Microcystin production of two airlift-grown cultures under a high light intensity and complete nutrient deprivation and the inoculation of a ceramic membrane. An immunological technique was used to elucidate where toxin production was greatest within a biofilm immobilized upon an externally skinless polyethersulfone capillary membrane. The externally skinless polyethersulfone capillary membrane was evaluated to assess homogeneity and the physical differences between membrane batches that led to the erratic, incomplete biofilm formation, as a biofilm of a constant thickness could not be immobilized. Microcystis aeruginosa PCC7806 was exposed to a variety of solvents in order to permeabilize the cyanobacteria, as that would have enabled a truly continuous extraction process for the metabolite. FDA hydrolysis had to be optimized in order to use it as an indicator of cell viability. In addition a single-step extraction of Microcystin was attempted using live bacteria. A capillary membrane module, containing the externally skinless polyethersulfone capillary membrane, inoculated using pressure filtration, was the most efficient combination to establish a biofilm. Cells that were no longer actively dividing and that lacked buoyancy displayed superior immobilization to cells that were actively dividing and buoyant. The immobilized cells did produce Microcystin but in much lower concentrations to cells grown in an airlift culture. Biofilms grown with a higher nitrate concentration, a lower biofilm thickness and a lower light intensity had a higher specific microcystin content, while biofilms with a higher nitrate concentration a lower light intensity and a longer growth period displayed the a greater toxin production per mm2 of membrane. Microcystin occurred at its highest concentration in cells just above the pore opening. The diffusion of nutrients occurred relatively quickly to the outside layers of the biofilm, with a true gradient being established laterally from these nutrient veins that were above the pores. Permeabilization of the cells proved unsuccessful, as cells that remained viable did not release the intracellular compound into the surrounding medium.
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- Date Issued: 2002