Seasonal and diurnal variation of nitrogenase activity (acetylene reduction) in barrel medic (Medicago truncatula Gaertn.) grown in pots

1978 ◽  
Vol 29 (5) ◽  
pp. 951 ◽  
Author(s):  
JJ Ruegg ◽  
AM Alston

Seasonal and diurnal variation of nitrogenase activity in Medicago truncatula Gaertn. was measured by means of the acetylene reduction assay on plants grown in pots. In a glasshouse set at 20°C, the seasonal pattern of acetylene reduction (AR) activity was closely correlated with dry weight and photosynthetic area. Short-term fluctuations in AR activity were mainly associated with irradiance. Measurements made of the diurnal variation of AR activity showed that rates of AR at noon were 10–60% (average 33%) higher than the mean daily rates. Effects of defoliation and shading gave further evidence for the importance of light and recent photosynthate for nitrogen fixation in root nodules of legumes. Values for acetylene reduction integrated over time were highly correlated with the total amount of nitrogen in the plant. The molar ratio of acetylene reduced to nitrogen accumulated by the plants at the end of the experiment was 1.2 : 1. The significance of this value is discussed. Multiple use of the same plant material to study the time course of nitrogen fixation by the AR assay was found to be feasible under certain conditions.


1977 ◽  
Vol 34 (3) ◽  
pp. 416-420 ◽  
Author(s):  
M. L. Guerinot ◽  
W. Fong ◽  
D. G. Patriquin

Sea urchins feeding in different macrophyte zones in St. Margaret's Bay, Nova Scotia, in November and December 1975 were examined for nitrogenase activity using the acetylene reduction technique. For sea urchins feeding on Laminaria digitata and L. longicruris, the average rate of acetylene reduction was equivalent to N2 fixation of 55 μg N2 per sea urchin per day assuming a 3:1 molar ratio of C2H2 reduction to N2 fixation. Lower nitrogenase activities were observed for sea urchins feeding on Agarum cribrosum and Zostera marina, and no nitrogenase activity was observed for sea urchins taken from bare rock substrate on which kelp had been absent for 4 yr. Samples of separated digestive tracts but not the degutted bodies of sea urchins exhibited nitrogenase activity, suggesting that the digestive tract is the site of nitrogen fixation in whole sea urchins.



1990 ◽  
Vol 17 (5) ◽  
pp. 489 ◽  
Author(s):  
Herdina ◽  
JH Silsbury

Methods of conducting acetylene reduction (AR) assay were appraised for estimating the nitrogenase activity of nodules of faba bean (Vicia faba L.). Factors considered were: (i) disturbance of plants when removing the rooting medium; (ii) assay temperature; (iii) the use of whole plants rather than detached, nodulated roots; (iv) diurnal variation in nodule activity; and (v) a decline in C2H4 production after exposure to C2H2. Plants growing in jars of 'oil dry' (calcined clay) had the same AR activity when assayed in situ in a closed system as when assayed after removal of the rooting medium. Assay temperatures of 12.5, 17.5 and 22.5°C influenced the specific rate of AR with the optimum at 17.5°C. Removal of the shoot resulted in a rapid decrease in AR activity in both vegetative and reproductive plants but the effect was much larger in the latter. AR and respiration by nodulated roots were closely linked and both varied markedly over a diurnal 12 h/12 h cycle. Since no fluctuation was found after nodules were detached, diurnal variation in the respiration of nodulated roots is attributed to change in nodule activity. Half of the dark respiration of nodulated roots was associated with respiration of the nodules and thus largely with N2 fixation. Since the AR assay provides no information on how electron flow in vivo is partitioned between reduction of N2 and reduction of protons, diurnal variation in hydrogen evolution (HE) in air and Ar/O2 in an open system was used to estimate this partitioning. Diurnal variation in apparent N2 fixation estimated in this manner was examined at a 'low' PPFD (300 μmol m-2 s-1) and at 'high' (1300 μmol m-2 s-1) to explore whether variation could be attributed to change in carbohydrate supply. Although HE in air and in Ar/O2 were both closely linked with the respiration of the nodulated root, apparent N2 fixation showed only a slight diurnal variation at 'low' light and almost none at 'high'. Vegetative plants showed no C2H2-induced decline in activity with exposure to C2H2 but reproductive plants did. This difference appears to be an age effect rather than attributable to flowering per se, since a decline occurred even when plants were kept vegetative by disbudding. A closed system for AR assay appears satisfactory for vegetative faba bean but such an assay over a 40-min period during the reproductive stage would underestimate nitrogenase activity by about 20%.



1981 ◽  
Vol 27 (1) ◽  
pp. 52-56 ◽  
Author(s):  
L. V. Wood ◽  
R. V. Klucas ◽  
R. C. Shearman

Turfs of 'Park' Kentucky bluegrass reestablished in the greenhouse and inoculated with Klebsiella pneumoniae (W6) showed significantly increased nitrogen fixation (acetylene reduction) compared with control turfs. Mean ethylene production rates per pot were 368 nmol h−1 for K. pneumoniae treated turfs, 55 nmol h−1 for heat-killed K. pneumoniae treated turfs, and 44 nmol h−1 for untreated turfs. Calculated lag periods before activity was observed were generally very short (less than 1 h).When 'Park' Kentucky bluegrass was grown from seed on soil-less medium of Turface, a fired aggregate clay, inoculation with K. pneumoniae (W6) resulted in 9 of 11 turfs showing nitrogenase activity (mean ethylene producion rate per pot was 195 nmol h−1). Only 3 of 11 turfs treated with heat-killed K. pneumoniae showed any activity and their mean rate of ethylene production (40 nmol h−1 per pot) was significantly lower than that for turfs treated with K. pneumoniae.Using the 'Park'–Turface soil-less model system it was shown that acetylene reducing activity was (i) root associated, (ii) generally highest at a depth of 1–4 cm below the surface, (iii) enhanced by washing excised roots, and (iv) inhibited by surface sterilization of excised roots. Klebsiella pneumoniae was recovered from Turface and roots showing acetylene reducing activity.



1979 ◽  
Vol 57 (11) ◽  
pp. 1285-1291 ◽  
Author(s):  
M. T. Ogan

The potential for nitrogen fixation in the rhizosphere and habitat of natural stands of Zizania aquatica (L) was studied by the acetylene reduction method. The data obtained suggested that this potential exists in the water column, the rhizosphere soil of the wild rice habitat, and on the root surfaces of the plants.In situ determination of rates of nitrogen fixation in the water column showed low but significant levels only in late spring – early summer and the rate was thought to be dependent on the presence of the blue-green algae Aphanizomenon. Laboratory experimental evidence showed that acetylene reduction by rhizosphere surface soil was attributable to Oscillatoria species while bacteria were more active in the subsurface soil and on the root surfaces. The bacteria-mediated nitrogenase activity was often preceded by a long lag period. The heterotrophic bacteria involved were enumerated, isolated, and characterised and they belong to the genera Azotobacter and Clostridium. Algal components of blooms occurring within the stands of Zizania at various times were identified.



2007 ◽  
Vol 85 (10) ◽  
pp. 1003-1006 ◽  
Author(s):  
P. D. Crittenden ◽  
X. Llimona ◽  
L. G. Sancho

Diurnal variation in N2-fixation (acetylene reduction) rate was measured in Thyrea girardii (Durieu & Mont.) Bagl. & Carestia and Thyrea confusa Henssen, lichens containing a unicellular cyanobacterial photobiont. In field assays, mean acetylene-reduction rates in the light were 25.8 ± 8.9 (n = 11) and 21.0 ± 5.6 nmol C2H4·g–1·h–1 (n = 13) for T. girardii and T. confusa, respectively, and the respective mean rates in the dark were 8.2 ± 1.8 (n = 26) and 13.5 ± 5.4 (n = 8) nmol C2H4·g–1·h–1. In laboratory assays under relatively isothermal conditions (ca. 19–22 °C), the maximum acetylene reduction rate (52.0 ± 6.0 nmol C2H4·g–1·h–1) was recorded in the light and the minimum rate (20.2 ± 6.0 nmol C2H4·g–1·h–1) in the dark. This diurnal pattern is contrary to expectations for unicellular cyanobacteria. We suggest carbon flow to the fungal symbiont reduces the potential for nitrogenase activity in the dark.



1983 ◽  
Vol 40 (11) ◽  
pp. 1853-1860 ◽  
Author(s):  
B. G. Brownlee ◽  
T. P. Murphy

Nitrogen fixation by Aphanizomenon flos-aquae in a prairie lake in southwestern Manitoba was dependent on the light intensity and in situ oxygen concentrations. The mean molar ratio of acetylene reduction to nitrogen reduction was 5.8:1. High external ammonium concentrations did not appear to inhibit nitrogen fixation over the short term. Nitrogen fixation was not directly initiated by the bloom collapse. We propose that the coupled sequence of ammonia volatilization and nitrogen fixation was triggered by the bloom collapse and that the bloom collapse was caused by coprecipitation of orthophosphate with carbonates. 32PO4 turnover was most rapid during periods when the lake was opalescent, presumably due to carbonate precipitaton.



1992 ◽  
Vol 70 (8) ◽  
pp. 1537-1544 ◽  
Author(s):  
Kerstin Huss-Danell ◽  
Per-Olof Lundquist ◽  
Helene Ohlsson

N2 fixation by grey alder, Alnus incana (L.) Moench, was studied in the field during two growing seasons in northern Sweden. Alders were planted in a nitrogen-poor soil. Each alder had its root system enclosed in an open-ended cylinder that was closed with a gas-tight lid around the stem base to serve as cuvette during nitrogenase activity (acetylene reducing activity) measurements. To follow the seasonal variation, nitrogenase activity was measured at noon on 15 occasions for each alder in 1987 and on 15 occasions in 1988. Diurnal variation in nitrogenase activity was studied at six occasions, but no obvious pattern in the diurnal variation was found. Nitrogenase activity began shortly after leaf emergence at the very end of May, increased in June, stayed high although with some variation through July and August, declined during September, and was zero in early October. Cumulative nitrogenase activity over the season was converted to cumulative N2 fixation after determination of molar ratio nitrogenase activity to N2 fixation. This conversion was facilitated as the Frankia chosen as symbiont was lacking hydrogenase activity. Control experiments showed that the introduced symbiont was the only infective Frankia in the soil. N2 fixation was estimated to be 0.23 and 2.83 g N/(alder∙year) in the 1st and 2nd year, respectively. Despite its young age, A. incana was apparently capable of high N2 fixation rates at the high latitude studied. Key words: Alnus incana, hydrogenase, intact plants, N2 fixation, seasonal variation, spreading of Frankia.



1975 ◽  
Vol 55 (2) ◽  
pp. 273-276 ◽  
Author(s):  
Larry N. Vanderhoef ◽  
Paul J. Leibson ◽  
Robert J. Musil ◽  
Chi-Ying Huang ◽  
Robert E. Fiehweg ◽  
...  


1979 ◽  
Vol 57 (1) ◽  
pp. 11-16 ◽  
Author(s):  
John Tjepkema

The acetylene reduction method was used to measure nitrogen fixation in soil cores of 16 cm diameter and 16 cm depth that included A and B horizon soil, roots, and decaying litter. Forty-three combinations of location and associated tree species were sampled. The rate of nitrogen fixation for most soil cores was 1 g N ha−1 day−1 or less, which extrapolates to less than 0.2 kg N ha−1 year−1. The highest rates, with values of up to 23 g N ha−1 day−1, were observed in old fields being invaded by trees. The time course of acetylene reduction was usually linear for a 24-h period, most of the activity was in the upper 15 cm of soil, and the maximum rates were observed in midsummer. No significant nitrogen fixation was observed in preliminary measurements of decaying woody litter or of aerial surfaces of trees and rocks. It is suggested that nitrogen fixation is not a significant input of nitrogen for the forests studied. If so, there may be significant unrecognized nitrogen inputs to forests, such as dry absorption of ammonia from the atmosphere.



1977 ◽  
Vol 30 (6) ◽  
pp. 593 ◽  
Author(s):  
IC MacRae

Acetylene reduction and nitrogen fixation by strains of Beijerinckia indica and B. lacticogenes increased with increased partial pressures of acetylene and nitrogen up to 80 kPa. The optical emission spectrophotometric method was used for the determination of 14N : 15N ratios. The molar ratios of acetylene to nitrogen varied greatly from the theoretical value.



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