The acetylene reduction technique as an assay for nitrogenase activity in the methane oxidizing bacterium Methylococcus capsulatus strain bath

1976 ◽  
Vol 109 (1-2) ◽  
pp. 147-151 ◽  
Author(s):  
Howard Dalton ◽  
Roger Whittenbury
Keyword(s):  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Reduction Technique ◽  
Methylococcus Capsulatus

A survey of angiosperms in Nova Scotia for rhizosphere nitrogenase (acetylene-reduction) activity

1978 ◽  
Vol 56 (18) ◽  
pp. 2218-2223 ◽  
Author(s):  
D. Smith ◽  
D. G. Patriquin
Keyword(s):  
Nova Scotia ◽  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Reduction Technique ◽  
Acetylene Reduction Activity ◽  
Forage Grasses ◽  
Reduction Activity ◽  
Tropical Forage ◽  
Poorly Drained Soils

Excised root samples from 901 plants, representing 130 species of nonnodulated angiosperms largely in upland, pioneering habitats, were assayed for nitrogenase activity by the acetylene-reduction technique after overnight preincubation of the samples under low pO2. Most samples and most species exhibited very low excised root acetylene-reducing activities, but for 19 species, maximum values were greater than 50 nmol C2H4∙g−1∙h−1. In situ C2H2 assays, conducted on 10 species which had exhibited maximum excised root activities greater than 10 nmol C2H4∙g−1∙h−1, indicated average belowground N2-fixation rates of 3 to 92 g N∙ha−1∙day−1 and maxima greater than 100 g N∙ha−1∙day−1 for 3 of the 10 species. The highest values were for grasses characteristic of poorly drained soils and for some dicotyledonous weeds. It is concluded that the potential of temperate-zone angiosperms for nitrogenase activity by 'associative symbioses' approaches that of tropical forage grasses.

Nitrogenase activity during chinook snowmelt sequences by Nostoc commune in Stipa–Bouteloa grassland

1983 ◽  
Vol 29 (8) ◽  
pp. 938-944 ◽  
Author(s):  
D. S. Coxson ◽  
K. A. Kershaw
Keyword(s):  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Reduction Technique ◽  
Ambient Light ◽  
Nostoc Commune ◽  
Temperature Optimum ◽  
Active Radiation ◽  
Cumulative Activity ◽  
Southern Alberta ◽  
Field Activity

Inactivation of the nitrogenase enzyme in winter has been observed for many terrestrial cyanophytes of north temperate habitats. In southern Alberta frequent winter chinook activity causes repeated snowmelt sequences. The pattern of nitrogenase activity in these periods has been examined (acetylene-reduction technique) over several snowmelt periods. Nostoc collected from snowmelt pockets was immediately assayed for nitrogenase activity under laboratory (20 °C, 200 E∙m−2s−1 photosynthetically active radiation (PAR)) and field (ambient light and temperature) conditions. Air and thallus temperatures for a typical snowmelt sequence rose from −20 to +5 and −5 to +20 °C, respectively, as chinook warming progressed. Activity on initial snowmelt emergence was 420 nmol C2H4∙g−1∙h−1, rising to 1050 nmol after 7 h, monitored in lab conditions. Field activity reached 90 nmol after 4 h, ceasing at 1800 as temperatures fell below 0 °C. Laboratory rates at morning thaw, day 2, were 1040 nmol, while field activity (3 °C, 900 μE) was 800 nmol. By midday laboratory rates reached 1198 nmol, while field rates (10 °C, 1500 μE) were 1077 nmol. Although these rates are well below those at the temperature optimum of 35 °C (6805 nmol), clearly no winter inactivation of the nitrogenase enzyme occurs here. Cumulative activity over winter chinook sequences may well exceed that occurring during infrequent summer hydration.

Estimating Nitrogenase Activity of Faba Bean (Vicia faba) by Acetylene Reduction (Ar) Assay

1990 ◽  
Vol 17 (5) ◽  
pp. 489 ◽  
Author(s):  
Herdina ◽  
JH Silsbury
Keyword(s):  
Diurnal Variation ◽  
Vicia Faba ◽  
Faba Bean ◽  
Closed System ◽  
N2 Fixation ◽  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Electron Flow ◽  
Specific Rate ◽  
Rooting Medium

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%.

Dinitrogen fixation and nodulation by Frankia in Alnus incana as affected by inorganic nitrogen in pot experiments with peat

1983 ◽  
Vol 61 (11) ◽  
pp. 2956-2963 ◽  
Author(s):  
U. Granhall ◽  
T. Ericsson ◽  
M. Clarholm
Keyword(s):  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Inorganic Nitrogen ◽  
Experimental Period ◽  
Alnus Incana ◽  
Nitrogen Application ◽  
Noticeable Effect ◽  
Essential Nutrients ◽  
Pot Experiments ◽  
Ph Level

The effects of single large or repeated, exponentially increasing applications of nutrients, with or without inorganic nitrogen and at two pH levels, on the growth, nodulation, acetylene reduction, and nutrient uptake in Alnus incana (L.) Moench were investigated in pot experiments with peat under controlled laboratory conditions. The repeated application of inorganic nitrogen did not suppress nitrogenase activity until the last 2 weeks, whereas an initial, large, nitrogen application effectively inhibited nodulation and activity throughout the 40-day experimental period. The mode of nitrogen application was thus found to be more important than the total amounts applied. Shoot length, leaf area, shoot–root relations, dry-matter production, and nitrogen contents of plants were determined at the end of the experiment, as well as the effect of Frankia inoculations. Nitrogenase activity was determined three times, at 0, 3, and 5 weeks. N2 fixation (balance/acetylene reduction) was found to be maximal, 55% of total nitrogen uptake, in minus-N pots with single applications of essential nutrients. The fastest growth was, however, noted in pots with single applications of all nutrients, including N. Among the latter, pots inoculated with Frankia showed the best growth, in spite of low nitrogenase activity. The only noticeable effect of a raised pH level was a reduced endophyte activity in minus-N pots with single applications of essential nutrients, due to increased N mineralization in the peat.

Nitrogen fixation (acetylene reduction) by Klebsiella pneumoniae in association with 'Park' Kentucky bluegrass (Poa pratensis L.)

1981 ◽  
Vol 27 (1) ◽  
pp. 52-56 ◽  
Author(s):  
L. V. Wood ◽  
R. V. Klucas ◽  
R. C. Shearman
Keyword(s):  
Nitrogen Fixation ◽  
Klebsiella Pneumoniae ◽  
Ethylene Production ◽  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Poa Pratensis ◽  
Kentucky Bluegrass ◽  
Surface Sterilization ◽  
Reducing Activity ◽  
Acetylene Reducing Activity

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.

scholarly journals Studies on nitrogenase activity of diazotrophic isolates from different rice production systems

2016 ◽  
Vol 8 (1) ◽  
pp. 284-289
Author(s):  
S. Kanimoli ◽  
K. Kumar
Keyword(s):  
Acetylene Reduction ◽  
Tamil Nadu ◽  
Production Systems ◽  
Nitrogenase Activity ◽  
Rice Production ◽  
Lowland Rice ◽  
Aerobic Rice ◽  
Marked Variation ◽  
Reduction Assay ◽  
First Time

The present study was carried out to evaluate the nitrogen fixing ability of diazotrophs isolated from the rhizosphere soils of rice which were grown in three different rice growing systems. A total of hundred and ten isolates obtained were subjected to Acetylene Reduction Assay (ARA) and ninety eight isolates recorded significant amount of nitrogenase activity in a range of 185.73 to 3794.55 nmoles of ethylene mg of protein-1 h-1. The highest nitrogenase activity was recorded by Derxia (3794.55 nmoles of ethylene mg of protein-1 h-1) isolated from Trichy (lowland). Among the three different rice production systems, isolates obtained from lowland rice (Derxia – 3794.5 nmoles of ethylene mg of protein-1 h-1) recorded higher nitrogenase activity followed by Aerobic (Pseudomonas - 2194.89 nmoles of ethylene mg of protein-1 h-1) and SRI (Azotobacter - 1971.85 nmoles of ethylene mg of protein-1 h-1) rice isolates. The results revealed marked variation in the ARA of the diazotrophic isolates obtained from lowland, SRI and Aerobic rice. The nitrogenase activity of diazotrophs from rice fields have been reported earlier but the nitrogenase activity of diazotrophs from three different rice production systems from various parts of Tamil Nadu is reported for the first time from India.

scholarly journals Factors Influencing Nitrogenase Activity (Acetylene Reduction) by Root Nodules of Groundnut, Arachis hypogaea L.

1983 ◽  
Vol 10 (1) ◽  
pp. 26-29 ◽  
Author(s):  
P. T. C. Nambiar ◽  
P. J. Dart
Keyword(s):  
Soil Moisture ◽  
Environmental Factors ◽  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Root Nodules ◽  
Activity Period ◽  
Diurnal Fluctuation ◽  
Factors Influencing ◽  
Arachis Hypogaea L ◽  
Reduction Assay

Abstract Acetylene reduction assay, used to measure nitrogenase activity of legume root nodules, is influenced by environmental factors, which limit its application. The effects of some of the environmental factors on acetylene reduction by groundnut root nodules are described. The activity was nonlinear during the first hour of incubation. Assay temperature above 25 C decreased the activity. Washing the nodulated roots prior to the assay also decreased the activity. The activity was influenced by light intensity, soil moisture, and moisture content in the incubation bottle. Diurnal fluctuation with one maximum and one minimum activity period during a 24 hour cycle was observed. Nitrogenase activity was higher during the postrainy season compared to that of the rainy season. A virginia cultivar Kadiri-71 had higher nitrogenase activity than a dwarf valencia cultivar, MH 2.

Potential for nitrogen fixation in the rhizosphere and habitat of natural stands of the wild rice Zizania aquatica

1979 ◽  
Vol 57 (11) ◽  
pp. 1285-1291 ◽  
Author(s):  
M. T. Ogan
Keyword(s):  
Nitrogen Fixation ◽  
Water Column ◽  
Acetylene Reduction ◽  
Wild Rice ◽  
Heterotrophic Bacteria ◽  
Nitrogenase Activity ◽  
Early Summer ◽  
Zizania Aquatica ◽  
Natural Stands ◽  
Root Surfaces

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.

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