The effect of light-dependent oxygen consumption on nitrogenase activity in Anabaena cylindrica

1986 ◽  
Vol 64 (9) ◽  
pp. 1843-1848 ◽  
Author(s):  
Donald L. Smith ◽  
David G. Patriquin ◽  
Margareta Dijak ◽  
George M. Curry
Keyword(s):  
Oxygen Consumption ◽  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Dark Respiration ◽  
Anabaena Cylindrica ◽  
Heat Treated ◽  
Carbonyl Cyanide ◽  
Photosynthetic Oxygen ◽  
Similar Decrease ◽  
Effect Of Light

Light-dependent oxygen consumption (LDOC) was observed in isolated heterocysts and in intact and sonicated CO2-fixing Anabaena cylindrica cells. The rate of LDOC in heterocysts was about three times that of CO2-fixing cells. Photosynthetic oxygen production by A. cylindrica became light saturated at 0.3 to 0.5 mW cm−2. LDOC and nitrogenase activity (acetylene reduction) increased with light intensity up to 2.5 mW cm−2 and incubation under air resulted in much larger relative acetylene reduction increases than incubation under N2. Carbonyl cyanide-m-chlorophenyl-hydrazone, 3-(3,4-dichlorophenyl)-1,1-dimethylurea, and cyanide did not affect the rate of LDOC in isolated heterocysts or cell-free preparations of CO2-fixing cells. However, all three substances induced LDOC in CO2-fixing cells. Heat treatment (100 °C for 1 min) caused a doubling of LDOC. Depletion of reduced carbon reserves by dark incubation caused a similar decrease in LDOC and dark respiration. The higher rates of LDOC observed in heat-treated materials were removed by catalase, but not by superoxide dismutase. Catalase injection released half of the O2 consumed through LDOC by heated preparations. LDOC increased with temperature up to 85 °C, and increased threefold with pH between pH 10 and 11.5. The possibility that LDOC may act to protect the nitrogenase of the heterocyst from oxygen inactivation is discussed.

Photorespiration and nitrogenase activity in the blue-green alga, Anabaena cylindrica

1972 ◽  
Vol 180 (1058) ◽  
pp. 87-102 ◽  
Keyword(s):  
Green Alga ◽  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Dark Respiration ◽  
Reducing Power ◽  
Blue Green Alga ◽  
Anabaena Cylindrica ◽  
Oxygen Inhibition ◽  
Highly Sensitive ◽  
Photosynthetic Oxygen

Oxygen uptake in the light (photorespiration) by the nitrogen-fixing blue-green alga Anabaena cylindrica may be up to twenty times the dark respiration rate. The rate of uptake in the light increases linearly with increasing p O 2 while dark respiration is saturated at a p O 2 near 0.05 atm. Photorespiration is inhibited rapidly and completely by DCMU (3 x 10 -5 m) but KCN (10 -4 m) has little effect. Exogenously supplied hydroxyethane sulphonate (10 -5 m), an inhibitor of glycollate oxidase activity, and glycollate do not affect respiration, although 14 C-labelled glycollate is assimilated in the light and in the dark. Photorespiration is highly sensitive to p CO 2 and to NaHCO 3 concentration and approaches true photosynthetic oxygen production at the CO 2 compensation point of 10 parts/10 6 . A CO 2 concentration of 0.02 atm completely inhibits photorespiration whereas true photosynthesis is scarcely affected. Conditions which stimulate photorespiration (low p CO 2 and high p O 2 ) progressively inhibit acetylene reduction. In short-term studies DCMU inhibits acetylene reduction under condi­tions which stimulate photorespiration but has little effect under conditions which inhibit photorespiration. The results suggest that photorespiration and nitrogenase activity com­pete indirectly for reducing power and that at least one mechanism of oxygen inhibition of nitrogenase activity is via a stimulation of photorespiration.

Reversibility of oxygen switch-off effect on Bacillus polymyxa nitrogenase

1991 ◽  
Vol 37 (10) ◽  
pp. 775-779 ◽  
Author(s):  
Hans-Paul Holzmann ◽  
Heinrich A. Berghammer ◽  
Martin Ortner ◽  
Kurt Haselwandter
Keyword(s):  
Oxygen Consumption ◽  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Cellular Respiration ◽  
Logarithmic Phase ◽  
Respiratory Protection ◽  
Bacillus Polymyxa ◽  
Closed Chamber ◽  
Oxygen Inhibition

The objective of this study was to analyse in vivo the effect of oxygen on the nitrogenase of Bacillus polymyxa. The culture technique employed in this study prevented spore formation by B. polymyxa during the entire period of exposure to acetylene. Under these conditions the acetylene-reduction assay allowed quantification of nitrogenase activity over long incubation periods (44 h). Nitrogenase activity was highest in cells harvested in the late logarithmic phase. At [Formula: see text] of 0.19 and 0.37 kPa, acetylene reduction was inhibited by 80 and 100%, respectively. This switch-off effect could be reversed through oxygen exhaustion, either by flushing the culture with N2 or by cellular respiration, suggesting a respiratory protection mechanism for the nitrogenase complex in B. polymyxa. Oxygen consumption measured by a closed-chamber respirometer showed a linear increase up to a [Formula: see text] of 0.2 kPa. Above 0.3 kPa a saturation in oxygen consumption was observed. Exposure to high oxygen pressures resulted in an irreversible loss of nitrogenase activity. The oxygen inhibition pattern was shown to be similar to that in other microaerophilic and anaerobic nitrogen-fixing microorganisms. Key words: Bacillus polymyxa, nitrogenase, oxygen inhibition, reversibility, respiration.

Nitrogen Fixation on a Tropical Volcano, La Soufrière (Guadeloupe): The Interaction of Temperature, Moisture, and Light with Net Photosynthesis and Nitrogenase Activity in Stereocaulon Virgatum and Response to Periods of Insolation Shock

1988 ◽  
Vol 20 (1) ◽  
pp. 63-81 ◽  
Author(s):  
R. P. Fritz-Sheridan ◽  
D. S. Coxson
Keyword(s):  
Quantum Yield ◽  
Light Intensity ◽  
Acetylene Reduction ◽  
Light Exposure ◽  
Nitrogenase Activity ◽  
Dark Respiration ◽  
Net Photosynthesis ◽  
High Light Intensity ◽  
High Light ◽  
Apparent Quantum Yield

AbstractThe response of net photosynthesis, dark respiration and acetylene reduction to temperature, moisture and light intensity were examined for Stereocaulon virgatum growing in the cloud/shroud zone on the tropical volcano La Soufrière, Guadeloupe, French West Indies. Rates for both acetylene reduction and net photosynthesis were maximal at saturating water contents, a pattern attributed to the finely branched nature of the phyllocladoid branchlets and the exposed position of spherical cephalodia, both of which minimize the formation of surface and interhyphal water films. Under conditions typical of those during cloud/shroud periods (13–16°C), thalli of S. virgatum exhibit many characteristics seen in other shade-tolerant lichen species. Net photosynthesis was light saturated at 300 μmol m−2 s−1 PAR, while the photocompensation point was less than 25 µmol m−2 s−1 PAR. Net photosynthetic uptake of carbon dioxide was optimal at 27–34°C, at which point light saturation was near 700 µmol m−2 s−1 PAR and the photocompensation point between 50 and 100 µmol m−2 s−1 PAR. Thalli of S. virgatin exhibited temperature-dependent sensitivity to high insolation. Only at 20°C were thalli able to tolerate high light exposure without reduction of apparent quantum yield. Exposure to high light intensity at 40°C inhibited the apparent quantum yield by almost 40% and acetylene reduction by 95%. This suggests brief periods of insolation shock may exert an influence disproportionately higher than either their frequency or duration. Thalli are normally exposed to cloud/shroud conditions but net photosynthetic uptake was maximal only during periods of elevated thallus temperature experienced at the onset of an insolation shock. However, with prolonged high insolation exposure and further elevation of thallus temperatures and thallus desiccation, severe impairment of subsequent photosynthetic activity ensues. S. virgatum may be characterized as a shade-tolerant species but its physiology is more adapted in some respects to conditions experienced during rare periods of full insolation.

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.

Mitochondrial respiratory control can compensate for intracellular O2 gradients in cardiomyocytes at low Po 2

2002 ◽  
Vol 283 (3) ◽  
pp. H871-H878 ◽  
Author(s):  
Eiji Takahashi ◽  
Koji Asano
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Diffusion ◽  
Respiratory Control ◽  
Oxygen Gradients ◽  
Nadh Fluorescence ◽  
Carbonyl Cyanide ◽  
Intrinsic Regulation ◽  
Electrical Pacing ◽  
Mitochondrial Oxidative Metabolism ◽  
Mitochondrial Respiratory

In isolated single cardiomyocytes with moderately elevated mitochondrial respiration, direct evidence for intracellular radial gradients of oxygen concentration was obtained by subcellular spectrophotometry of myoglobin (Mb). When oxygen consumption was increased by carbonyl cyanide m-chlorophenylhydrazone (CCCP) during superfusion of cells with 4% oxygen, Po 2 at the cell core dropped to 2.3 mmHg, whereas Mb near the plasma membrane was almost fully saturated with oxygen. Subcellular NADH fluorometry demonstrated corresponding intracellular heterogeneities of NADH, indicating suppression of mitochondrial oxidative metabolism due to relatively slow intracellular oxygen diffusion. When oxygen consumption was increased by electrical pacing in 2% oxygen, radial oxygen gradients of similar magnitude were demonstrated (cell core Po 2 = 2.6 mmHg). However, an increase in NADH fluorescence at the cell core was not detected. Because CCCP abolished mitochondrial respiratory control while it was intact in electrically paced cardiomyocytes, we conclude that mitochondria with intact respiratory control can sustain electron transfer with reduced oxygen supply. Thus mitochondrial intrinsic regulation can compensate for relatively slow oxygen diffusion within cardiomyocytes.

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