The acetylene-reduction assay was used for in situ and laboratory assessment of biological nitrogen fixation in the acidic, waterlogged, muskeg ecosystem of the southern James Bay area, in the region of Moosonee, Ontario. In situ assays and subsequent laboratory experimentation revealed that nitrogenase activity was predominately a function of the activities of heterocystic blue-green bacteria associated with surface water, with the phyllosphere of mosses, and with at least one lichen, a species of Peltigera. No such in situ activity was detected in the subsurface organic material, even when such material was amended with glucose. However, under laboratory conditions at 20 degrees C, nitrogenase activity was evident in the subsurface layers after an extended lag and was shown to be higher under anaerobic than under aerobic conditions, to have an optimum temperature range extending about a mean of 20 degrees C, and to be stimulated by glucose. This potential for subsurface nitrogen fixation proved to be related to the presence of microorganisms existing in anaerobic microsites within the organic layers and no microorganisms capable of fixation could be detected under aerobic incubation.
Arranged in descending order of nitrogen-fixing (acetylene-reducing) potential the sites examined were mesic meadow and peat polygon troughs (equal rank), transition zone between mesic meadow and gravel ridge, gravel ridge, polar dessert, and peat polygon tops. The dominant nitrogen-fixing microorganisms, as in other Arctic areas, were blue-green bacteria, especially those epiphytic on Arctic mosses. The epiphytic association exhibited an optimum temperature for fixation of 20 degrees C. Other bacteria potentially able to fix nitrogen were present in the soils examined but their activity was severely restricted by low soil temperatures and lack of readily utilizable energy sources. These bacteria included members of the genera Klebsiella (the most numerous), Bacillus, Clostridium, and Beijerinckia (scarce). Also present at many of the sites was an unidentified yellow-pigmented fixer which was not Mycobacterium flavum. All fixers were psychotrophic rather than psychrophilic, having an optimum temperature greater than 20 degrees C but capable of slow growth at 5 degrees C or lower. The rate of acetylene reduction by the epiphytic system increased with the number of successive exposures to acetylene, a phenomenon of some significance in any calculations designed to measure the amount of nitrogen fixed in certain ecosystems.
A hitherto undescribed species of yellow-pigmented, Gram-negative Clostridium sp., possessing nitrogenase activity, has been isolated from a number of sampling sites on the Truelove Lowland of Devon Island in the Canadian high Arctic. This bacterium, tentatively designated Clostridium arcticum sp. nov., accounted for 19% of all isolates recovered which were capable of anaerobic nitrogen fixation.
An apparently new species of Clostridium was isolated from mud samples obtained from a stagnant lake near Athens, Georgia. These organisms were able to utilize inorganic pyrophosphate as a source of energy for growth, a phenomenon representing the simplest ATP-generating system in the biological world.
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