Nitrogen fixation by Myrica gale root nodules Massachusetts wetland

Summary Nitrogenase activity was measured by acetylene reduction in excised Myrica gale nodules collected throughout the growing season at two sites associated with a small lake in central Massachusetts. One site was in an open, weakly minerotrophic peatland dominated by M. gale and the other was on the lakeshore. Nitrogenase activity appeared in late May when the leaves unfolded, reached a peak in July when the maximum number of leaves was present, and declined until it was no longer measurable in late October several days after all leaves had fallen. Summer activities were substantially higher at the peatland site than the lakeshore. Maximum activities were 19.8±1.9 and 8.1±0.7 mol/h x g dry weight (x±SE; N=20) at the peatland and lakeshore sites respectively.Nitrogenase activities were very low at 5° C and increased linearly from 10 to 30° C, the highest temperature examined. The maximum soil temperature measured was 20° C, and no significant diurnal fluctuation in activity was detected.Annual nitrogen fixation calculated from the seasonal nitrogenase activity curve was 34 kg N/ha x yr at the peatland site with mean dry weight nodule biomass of 104 kg/ha, and 24 kg N/ha x yr at the lakeshore with 111 kg/ha nodule biomass. These rates of nitrogen fixation are equivalent to 4–5x the amount of nitrogen contained in bulk precipitation and are major components in the nitrogen budgets of the M. gale plants and wetlands in which they grow.     

Significant nonsymbiotic nitrogen fixation in Patagonian ombrotrophic bogs

Nitrogen (N) nutrition in pristine peatlands relies on the natural input of inorganic N through atmospheric deposition or biological dinitrogen (N₂) fixation. However, N₂ fixation and its significance for N cycling, plant productivity, and peat buildup are mostly associated with the presence of Sphagnum mosses. Here, we report high nonsymbiotic N₂‐fixation rates in two pristine Patagonian bogs with diversified vegetation and natural N deposition. Nonsymbiotic N₂ fixation was measured in samples from 0 to 10, 10 to 20, and 40 to 50 cm depth using the ¹⁵N₂ assay as well as the acetylene reduction assay (ARA). The ARA considerably underestimated N₂ fixation and can thus not be recommended for peatland studies. Based on the ¹⁵N₂ assay, high nonsymbiotic N₂‐fixation rates of 0.3–1.4 μmol N₂ g^?1 day^?1 were found down to 50 cm under micro‐oxic conditions (2 vol.%) in samples from plots covered by Sphagnum magellanicum or by vascular cushion plants, latter characterized by dense and deep aerenchyma roots. Peat N concentrations point to greater potential of nonsymbiotic N₂ fixation under cushion plants, likely because of the availability of easily decomposable organic compounds and oxic conditions in the rhizosphere. In the Sphagnum plots, high N₂ fixation below 10 cm depth rather reflects the potential during dry periods or low water level when oxygen penetrates the top peat layer and triggers peat mineralization. Natural abundance of the ¹⁵N isotope of live Sphagnum (5.6 δ‰) from 0 to 10 cm points to solely N uptake from atmospheric deposition and nonsymbiotic N₂ fixation. A mean ¹⁵N signature of ?0.7 δ‰ of peat from the cushion plant plots indicates additional N supply from N mineralization. Our findings suggest that nonsymbiotic N₂ fixation overcomes N deficiency in different vegetation communities and has great significance for N cycling and peat accumulation in pristine peatlands.     

Consequences of Sporangial Development for Nodule Function in Root Nodules of Comptonia peregrina and Myrica gale

Frankia sp., the actinomycetous endophyte in nitrogen-fixing actinorhizal nodules, may differentiate two forms from its hyphae: vesicles and sporangia. In root nodules of Comptonia peregrina (L.) Coult. and Myrica gale L., sporangia may be either absent or present. Nitrogenase activity and symbiotic efficiency were contrasted in spore(+) and spore(−) nodules of these two host genera. Seedlings of C. peregrina nodulated with the spore(+) inoculum showed only 60% of the nitrogenase activity and 50% of the net size of their spore(−) counterparts after 12 weeks of culture. Measurements of acetylene reduction (i.e., nitrogenase activity) were coordinated with samplings of nodules for structural studies. Significant differences in acetylene reduction rates were discernible between spore(+) and spore(−) nodules commencing 4 weeks after nodulation, concomitant with the maturation of sporangia in the nodule. Spore(+) nodules ultimately reached less than half of the rate of nitrogenase activity of spore(−) nodules. Both types of nodules evolved only small amounts of molecular hydrogen, suggesting that both were equally efficient in recycling electrons lost to the reduction of hydrogen ions by nitrogenase. Respiratory cost of nitrogen fixation, expressed as the quotient of micromole CO₂ to micromole ethylene evolved by excised nodules, was significantly greater in spore(+) than in spore(−) nodules. M. gale spore(−) nodules showed variable effectivity, though all had low CO₂ to ethylene evolution ratios. M. gale spore(+) nodules resembled C. peregrina spore(+), with low effectivity and high respiratory cost for nitrogen fixation.     

Phosphorus regulation of nitrogen fixation in a traditional Mexican agroecosystem

Although nitrogen is considered to be the nutrient that most commonly limits production of natural and managed terrestrial ecosystems, I propose that phosphorus may regulate productivity in many continuously cultivated agroecosystems that do not receive applications of synthetic fertilizers. One way P may limit agroecosystem productivity is by controlling nitrogen fixation of legume crops, thus affecting nitrogen availability in the overall agroecosystem. I tested this hypothesis in two studies by examining the effect of phosphorus nutrition on nitrogen fixation of alfalfa in traditional Mexican agroecosystems. All farms used in the research relied on alfalfa as the primary nitrogen source for maize cultivation and other crops, and had minimal or no reliance on synthetic fertilizers.In one study, I used the natural abundance of¹⁵N to estimate nitrogen fixation in five alfalfa plots with soils representing a wide range of P fertility. I found a correlation of r = 0.85 between foliage P concentrations and nitrogen fixation in the alfalfa plots. Mean nitrogen fixation in alfalfa plots ranged between 232–555 kg ha^–1 yr^–1 as estimated by the¹⁵N-natural abundance method.In a second study, I sampled soils from alfalfa plots on traditional farms located in 5 different physiographic regions of Mexico. Half of each soil sample was augmented with phosphorus in a greenhouse experiment. I grew alfalfa on the fertilized and unfertilized soils from each site and then determined nitrogenase activity (acetylene reduction) of the Rhizobium on the plant roots. Nitrogenase activity increased in the alfalfa grown on all soils with added phosphorus, with two of the five differences being statistically significant at P < 0.01, 0 and one at P < 0.05. Foliage P concentrations and nitrogenase activity were 0 positively correlated (r = 0.81,P < 0.01).0     

Nitrogen Fixation (Acetylene Reduction) Associated with Decaying Leaves of Pond Cypress (Taxodium distichum var. nutans) in a Natural and a Sewage-Enriched Cypress Dome

Surface litter from a natural and a sewage-enriched cypress dome in north-central Florida showed a pronounced seasonal pattern of nitrogenase (acetylene reduction) activity associated with seasonal leaf fall from deciduous trees in the domes. Samples of peat from cores indicated negligible nitrogenase activity below the surface layer. Integrating the monthly rates of nitrogen fixation (based on the theoretical molar ratio of 3:2 for C₂H₄/NH₃) yielded 0.39 and 0.12 g of N/m² per year fixed in the litter of the natural and sewage-enriched domes, respectively. The nitrogen fixed in the first 3 months after leaf fall in the natural dome represented about 14% of the nitrogen increment in the decomposing cypress leaves, but fixation contributed a negligible amount of nitrogen (<1%) to decomposing litter in the sewage-enriched dome.     

Changes in Asymbiotic, Heterotrophic Nitrogen Fixation on Leaf Litter of Metrosideros polymorpha with Long-Term Ecosystem Development in Hawaii

We measured nitrogenase activity (acetylene reduction) of asymbiotic, heterotrophic, nitrogen-fixing bacteria on leaf litter from the tree Metrosideros polymorpha collected from six sites on the Hawaiian archipelago. At all sites M. polymorpha was the dominant tree, and its litter was the most abundant on the forest floor. The sites spanned a soil chronosequence of 300 to 4.1 million y. We estimated potential nitrogen fixation associated with this leaf litter to be highest at the youngest site (1.25 kg ha^-1 y^-1), declining to between 0.05 and 0.22 kg ha^-1 y^-1 at the oldest four sites on the chronosequence. To investigate how the availability of weathered elements influences N fixation rates at different stages of soil development, we sampled M. polymorpha leaf litter from complete, factorial fertilization experiments located at the 300-y, 20,000-y and 4.1 million–y sites. At the youngest and oldest sites, nitrogenase activity on leaf litter increased significantly in the plots fertilized with phosphorus and “total” (all nutrients except N and P); no significant increases in nitrogenase activity were measured in leaf litter from treatments at the middle-aged site. The results suggest that the highest rates of N fixation are sustained during the “building” or early phase of ecosystem development when N is accumulating and inputs of geologically cycled (lithophilic) nutrients from weathering are substantial. Received 4 February 1999; accepted 29 March 2000.     

Nitrogen-Fixing Activity in Peat Soils from a Raised Bog

The nitrogenase (acetylene reductase) activity in monolithic and minced peat samples was found to be low, no more than 0.014–0.022 mg N/(kg h). Incorporation of the ¹⁵N₂ isotope into organic compounds of peat soil was 2.71–8.13 mg N/kg over 15 days. The nitrogen-fixing activity was the highest in a 10- to 20-cm layer of soil and much lower in the upper (under green moss) and deeper (20- to 30-cm) layers. The addition of glucose to soil samples stimulated nitrogen fixation considerably after 18–26 h. The maximum nitrogenase activity (3.5–3.8 mg N/(kg h)), observed after 60–70 h, coincided with the peak of respiratory activity. A repeated addition of glucose after its exhaustion increased nitrogenase activity, without a lag period, to 8.5 mg N/(kg h). Investigation of the effect of environmental factors (temperature, pH, aeration, and light intensity) on potential nitrogen-fixing activity in peat samples revealed that nitrogen fixation could proceed in a wide range of pH values (from 3.0 to 7.5) and temperatures (from 5 to 35°C). The nitrogen-fixing bacteria belonging to different trophic groups were enumerated by using nitrogen-free media with pH values and mineralization levels close to those in situ. In samples of peat soil, diazotrophic methanol-utilizing bacteria prevailed (2.0–2.5 × 10⁶ cells/g); the second largest group was facultatively anaerobic bacteria of the family Enterobacteriaceae.     

Effect of spring flooding on endophyte differentiation,nitrogenase activity,root growth and shoot growth inMyrica gale

Summary Spring flooding was investigated as a possible limiting factor in the development of nitrogenase activity, root growth, and shoot growth inMyrica gale. Dormant, one year oldMyrica gale plants were placed in a greenhouse in early April and given three treatments: control (not flooded), flooded-water (flooded with water to 2.5 cm above the soil level) and flooded-peat (flooded with water-saturated peat to 4.0 cm above the soil level). Nitrogenase activity was absent at budbreak but appeared concurrently with the differentiation of vesicles by theFrankia sp. endophyte. Flooding delayed the onset of nitrogenase activity, substantially reduced the specific nitrogenase activity of the nodules, and also severely limited the production of the new nodule biomass. Consequently by 67 days past budbreak nitrogenase activity was much greater in the control plants (5.55±0.42 mol C₂H₄/plant.h; ± SE; N=9) than in the flooded-water (1.18±0.29) and flooded-peat (0.15±0.05) plants. Production of new secondary roots was substantially reduced in the flooded plants but adventitious roots were rapidly produced along the flooded portion of the stem in the better aerated zone near the surface. New nodules formed on several adventitious roots by 67 days indicating that the plants are able to replace their largely nonfunctional deeply flooded nodules with new nodules in the aerobic zone. Initially shoot growth was unaffected by flooding but by 67 days the flooded plants had substantially less leaf biomass, lower leaf and stem nitrogen concentrations, and less total shoot nitrogen content than the control plants.     

Dry deposition of ammonia gas drives species change faster than wet deposition of ammonium ions: evidence from a long‐term field manipulation

Although the effects of atmospheric nitrogen deposition on species composition are relatively well known, the roles of the different forms of nitrogen, in particular gaseous ammonia (NH₃), have not been tested in the field. Since 2002, we have manipulated the form of N deposition to an ombrotrophic bog, Whim, on deep peat in southern Scotland, with low ambient N (wet + dry = 8 kg N ha^?1 yr^?1) and S (4 kg S ha^?1 yr^?1) deposition. A gradient of ammonia (NH₃, dry N), from 70 kg N ha^?1 yr^?1 down to background, 3–4 kg N ha^?1 yr^?1 was generated by free air release. Wet ammonium (NH₄⁺, wet N) was provided to replicate plots in a fine rainwater spray (NH₄Cl at +8, +24, +56 kg N ha^?1 yr^?1). Automated treatments are coupled to meteorological conditions, in a globally unique, field experiment. Ammonia concentrations were converted to NH₃‐N deposition (kg N ha^?1) using a site/vegetation specific parameterization. Within 3 years, exposure to relatively modest deposition of NH₃, 20–56 kg NH₃‐N ha^?1 yr^?1 led to dramatic reductions in species cover, with almost total loss of Calluna vulgaris, Sphagnum capillifolium and Cladonia portentosa. These effects appear to result from direct foliar uptake and interaction with abiotic and biotic stresses, rather than via effects on the soil. Additional wet N by contrast, significantly increased Calluna cover after 5 years at the 56 kg N dose, but reduced cover of Sphagnum and Cladonia. Cover reductions caused by wet N were significantly different from and much smaller than those caused by equivalent dry N doses. The effects of gaseous NH₃ described here, highlight the potential for ammonia to destroy acid heathland and peat bog ecosystems. Separating the effects of gaseous ammonia and wet ammonium deposition, for a peat bog, has significant implications for regulatory bodies and conservation agencies.     

Sources,fates, and impacts of nitrogen inputs to terrestrial ecosystems: review and synthesis

The relative importance of nitrogen inputs from atmospheric deposition and biological fixation is reviewed in a number of diverse, non-agricultural terrestrial ecosystems. Bulk precipitation inputs of N (l–l2 kg N ha^–1 yr^–1) are the same order of magnitude as, or frequently larger than, the usual range of inputs from nonsymbiotic fixation (< 1=" –=" 5=" kg=" n=">^–1 yr^–1), especially in areas influenced by industrial activity. Bulk precipitation measurements may underestimate total atmospheric deposition by 30–40% because they generally do not include all forms of wet and dry deposition. Symbiotic fixation generally ranges from 10–160 kg N ha^–1 yr^–1) in ecosystems where N-fixing species are present during early successional stages, and may exceed the range under unusual conditions.Rates of both symbiotic and nonsymbiotic fixation appear to be greater during early successional stages of forest development, where they have major impacts on nitrogen dynamics and ecosystem productivity. Fates and impacts of these nitrogen inputs are important considerations that are inadequately understood. These input processes are highly variable in space and time, and few sites have adequate comparative information on both nitrogen deposition and fixation.

Effects of available P and N:P ratios on non-symbiotic dinitrogen fixation in tallgrass prairie soils

Summary Prescribed burning is a major control over element cycles in Tallgrass prairie (Eastern Kansas, USA). In this paper we report potential effects of fire on nonsymbiotic nitrogen fixation. Fire resulted in additions of available P in ash, which may stimulate nitrogen fixation by terrestrial cyanobacteria. Cyanobacterial nitrogenase activity and biomass responded positively to additions of ash or P in laboratory assays using soil. Further assays in soil showed that cyanobacteria responded to changes in available N:available P ratio (aN:P) across a range of concentrations. Nitrogen fixation rate could be related empirically to aN:P via a log-linear relationship. Extrapolation of laboratory results to the field yielded a maximal estimate of 21 kg N ha^-1 y^-1. Results support arguments from the marine and terrestrial literature that P availability is central to regulation of ecosystem N budgets.     

Sensitivity of C Sequestration in Reintroduced Sphagnum to Water-Level Variation in a Cutaway Peatland

The reintroduction of Sphagnum fragments has been found to be a promising method for restoring mire vegetation in a cutaway peatland. Although it is known that moisture controls Sphagnum photosynthesis, information concerning the sensitivity of carbon dynamics on water‐level variation is still scarce. In a 4‐year field experiment, we studied the carbon dynamics of reintroduced Sphagnum angustifolium material in a restored (rewetted) cutaway peatland. Cutaway peatland restored by Sphagnum reintroduction showed high sensitivity to variation in water level. Water level controlled both photosynthesis and respiration. Gross photosynthesis (P_G) had a unimodal response to water‐level variation with optimum level at ?12 cm. The range of water level for high P_G (above 60% of the maximum light‐saturated P_G) was between 22 and 1 cm below soil surface. Water level had a dual effect on total respiration. When the water level was below soil surface, peat respiration increased rapidly along the lowering water level until the respiration rate started to slow down at approximately ?30 cm. Contrary to peat respiration, the response of Sphagnum respiration to water‐level variation resembled that of photosynthesis with an optimum at ?12 cm. In optimal conditions, Sphagnum reintroduction turned the cutaway site from carbon source to a sink of 23 g C/m² per season (mid‐May to the end of September). In dry conditions, lowered photosynthesis together with the higher peat respiration led to a net loss of 56 g C/m². Although the water level above the optimum amplitude restricted CO₂ fixation, a decrease in peat respiration led to a positive CO₂ balance of 9 g C/m².     

Comparison of carbon and nitrogen accumulation rate between bog and fen phases in a pristine peatland with the fen-bog transition

Long-term carbon and nitrogen dynamics in peatlands are affected by both vegetation production and decomposition processes. Here, we examined the carbon accumulation rate (CAR), nitrogen accumulation rate (NAR) and δ¹³C, δ¹⁵N of plant residuals in a peat core dated back to ~8500 cal year BP in a temperate peatland in Northeast China. Impacted by the tephra during 1160 and 789 cal year BP and climate change, the peatland changed from a fen dominated by vascular plants to a bog dominated by Sphagnum mosses. We used the Clymo model to quantify peat addition rate and decay constant for acrotelm and catotelm layers during both bog and fen phases. Our studied peatland was dominated by Sphagnum fuscum during the bog phase (789 to −59 cal year BP) and lower accumulation rates in the acrotelm layer was found during this phase, suggesting the dominant role of volcanic eruption in the CAR of the peat core. Both mean CAR and NAR were higher during the bog phase than during the fen phase in our study, consistent with the results of the only one similar study in the literature. Because the input rate of organic matter was considered to be lower during the bog phase, the decomposition process must have been much lower during the bog phase than during the fen phase and potentially controlled CAR and NAR. During the fen phase, CAR was also lower under higher temperature and summer insolation, conditions beneficial for decomposition. δ¹⁵N of Sphagnum hinted that nitrogen fixation had a positive effect on nitrogen accumulation, particular in recent decades. Our study suggested that decomposition is more important for carbon and nitrogen sequestration than production in peatlands in most conditions and if future climate changes or human disturbance increase decomposition rate, carbon sequestration in peatlands will be jeopardized.     

Effect of methabenzthiazuron on growth and nitrogenase activity in Vicia faba

The effects of the herbicide methabenzthiazuron (175 and 220 g ha^-1) on vegetative and reproductive growth, nodulation and nitrogenase activity of Vicia faba were studied in the field under Mediterranean conditions. Nitrogenase activity of excised nodules was estimated using the acetylene reduction assay four times during the developmental period. Leaf area index, dry weight and nitrogen content of the different parts of the plants were measured. Methabenzthiazuron-treated plants showed an increase in nodulation, nitrogenase activity and vegetative growth at early pod fill. Methabenzthiazuron also caused an increase in leaf N content and fruits. These were transient effects found during early and mid pot fill. Nevertheless, plants treated with these sublethal doses of herbicide improved seed production and nitrogen content of seeds at harvest time. The stimulatory effect of methabenzthiazuron on N₂ fixation and vegetative growth seems not be related with the transient stimulatory effect on photosynthetic capacity, also caused by the herbicide, since the stimulatory effect on N₂ fixation was apparent during pod fill, when photosynthetic capacity declined and was not modified by methabenzthiazuron.     

NITROGEN FIXATION ASSOCIATED WITH NATURAL AND CULTIVATED STANDS OF TYPHA LATIFOLIA L. (TYPHACEAE)

Acetylene reduction by bacteria associated with Typha latifolia L. roots and rhizomes was studied in the field and in the laboratory. In situ studies indicated that the rate of acetylene reduction in a natural cattail population was ca. 4-fold higher than in a cultivated cattail stand. Nitrogenase activity was found to occur principally in the rhizosphere of roots and rhizomes with the greatest activity occurring in association with mature roots. Scanning electron and light microscopy, and 2, 3, 5 triphenyltetrazolium chloride reduction showed that bacterial associations were limited to the rhizoplane of this angiosperm. One diazotrophic bacterial genus was found to be associated with roots and rhizomes and was identified as the facultative anaerobe Bacillus. Contribution of free-living cyanobacteria to total nitrogen fixation in the natural stand was negligible. Calculations show that a natural stand of cattails may fix 18 kg nitrogen ha^−-1 yr^−-1 or ca. 8.2% of the total nitrogen present in the standing crop.     

Nitrogen fixation in a Mexican coffee plantation

Fertilizer studies in Mexico indicate that coffee production can be stimulated by added nitrogen. One traditional method of coffee cultivation employs leguminous trees for shade, but these species may also play an important role in coffee production by biologically fixing nitrogen. The presence and importance of nitrogen fixation was evaluated in four systems: coffee only, coffee plus the leguminous shade treeInga jinicuil Schletchter, coffee plus the leguminous treeInga vera H.B. and K., and coffee plus banana and orange trees. In all systems coffee leaves with epiphylls, wood litter, soil, roots, and root nodules were assayed for nitrogen fixing activity with the acetylene reduction technique. All components of these systems exhibited activity except roots. Total apparent fixation was highest in theInga jinicuil site, and equivalent to >40 kg N ha^?1 yr^?1 assuming a 3∶1 C₂H₂∶N₂ ratio. The activity was primarily associated withInga jinicuil nodules. Apparent fixation in the other three sites was less than 1 kg N ha^?1 yr^?1. Nitrogen fixed in theI. jinicuil site was 53% of the average amount of fertilizer nitrogen applied annually, suggesting that fixation by non-crop legumes can be an important nitrogen source for coffee agro-ecosystems.     

Nitrogen fixation by Nodularia spumigena Mertens (Cyanobacteriaceae). 2: Laboratory studies

The effects of changes in diurnal light patterns, salinity, and phosphorus on nitrogen fixation (as measured by acetylene reduction) by Nodularia spumigena Mertens were examined. As well, the effects of added inorganic nitrogen on growth, nitrogen fixation and heterocyt frequencies, and changes in nitrogen fixation and heterocyst frequencies during the growth cycle of Nodularia in cultures were determined.The diurnal pattern of nitrogenase activity in Nodularia was primarily light-induced, though dark activity did occur. Nitrogenase activity following a period of darkness exceeded the normal light rate (> 90 compared to 50 nmol · C₂H₂ reduced · ml^–1 · h^–1). Nitrogen fixation was reduced by high and very low salinities (5 to 10 was the optimum range), and added phosphorus stimulated nitrogenase in P-starved cells. Added nitrogen (ammonium or nitrate) had no effect on the growth of Nodularia, but in short term studies, ammonium completely inhibited nitrogenase activity. Heterocyst frequencies were greatest in the log phase of growth (to 40 per mm). During stationary phase, nitrogenase activity was negligable.     

Growth,nitrogen accumulation,and symbiotic dinitrogen fixation in pure and mixed plantings of hybrid poplar and black alder

Summary Growth and N accumulation were assessed in pure and mixed plantings (2 years old) of hybrid poplar and black alder in southern Québec. Symbiotic dinitrogen fixation was evaluated by natural¹⁵N dilution. Growth of hybrid poplar plants and N accumulation in their tissues increased with their decreasing contribution to species ratio whereas no differences among treatments were measured for black alder. Yield and N content per hectare of aboveground components increased with the proportion of black alder in the plantation. Symbiotic dinitrogen fixation was estimated at 68% of alder nitrogen in both pure and mixed treatments. The maximum rate of N-fixation was 53kg ha^–1 yr^–1 in pure alder plots. The amount of nitrogen accumulated in entire plants of black alder from symbiotic fixation could be sufficient to balance the N export in harvested stems and branches of short-rotation plantations containing at least 33% of alder.     

Growth and N2 fixation in an Alnus hirsuta (Turcz.) var. sibirica stand in Japan

To estimate the N₂ fixation ability of the alder (Alnus hirsuta (Turcz.) var. sibirica), we examined the seasonal variation in nitrogenase activity of nodules using the acetylene reduction method in an 18-year-old stand naturally regenerated after disturbance by road construction in Japan. To evaluate the contribution of N₂ fixation to the nitrogen (N) economy in this alder stand, we also measured the phenology of the alder, the litterfall, the decomposition rate of the leaf litter, and N accumulation in the soil. The acetylene reduction activity per unit nodule mass (ARA) under field conditions appeared after bud break, peaked the maximum in midsummer after full expansion of the leaves, and disappeared after all leaves had fallen. There was no consistent correlation between ARA and tree size (dbh). The amount of N₂ fixed in this alder stand was estimated at 56.4 kg ha^?1 year^?1 when a theoretical molar ratio of 3 was used to convert the amount of reduced acetylene to the amount of fixed N₂. This amount of N₂ fixation corresponded to the 66.4% of N in the leaf litter produced in a year. These results suggested that N₂ fixation still contributed to the large portion of N economy in this alder stand.     

Nitrogen fixation by the benthic freshwater cyanobacterium Lyngbya wollei

The ability of the benthic cyanobacterium Lyngbya wollei to fix nitrogen was studied using field samples and axenic cultures. L. wollei was collected and isolated from Lake Okeechobee, Florida, where it forms extensive mats. Rates of acetylene reduction up to 39.1 nmol mg dry wt⁻¹ h⁻¹ were observed for field samples. The maximum observed rate of acetylene reduction in axenic laboratory cultures was 200 nmol mg dry wt⁻¹ h⁻¹. Aerobic conditions limited nitrogen fixation activity, but dark/light cycles promoted the development of activity. Reduced oxygen levels appeared to be required for the development of significant levels of nitrogenase activity. The level of irradiance also had a significant impact on the level of activity. The potential significance of nitrogen fixation to Lyngbya production is discussed.