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

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 25 (10) ◽  
pp. 1197-1200 ◽  
Author(s):  
R. C. Shearman ◽  
W. L. Pedersen ◽  
R. V. Klucas ◽  
E. J. Kinbacher

Associative nitrogen fixation in Kentucky bluegrass (Poa pratensis L.) turfs inoculated with five nitrogen-fixing bacterial isolates was evaluated using the acetylene reduction assay and nitrogen accumulation as indicators of fixation. 'Park' and 'Nugget' Kentucky bluegrass turfs were grown in controlled environment chambers and inoculated with Klebsiella pneumoniae (W-2, W-6, and W-14), Erwinia herbicola (W-8), and Enterobacter cloacae (W-11). 'Park' inoculated with K. pneumoniae (W-6) had significant acetylene reduction activity using undisturbed turfs. Other treatments including turfs treated with heat-killed cells had no significant difference in acetylene reduction. In a second study, 'Park' and 'South Dakota Certified' turfs were grown in a greenhouse and inoculated with K. pneumoniae (W-6) and E. herbicola (W-8). 'Park' inoculated with K. pneumoniae (W-6) had increased acetylene reduction activity rates and also a greater nitrogen accumulation in aerial tissues when compared to controls. Acetylene reduction activity was correlated (r = 0.92) to nitrogen accumulation. Other treatments did not effectively increase acetylene reduction activity or nitrogen accumulation.


1974 ◽  
Vol 139 (1) ◽  
pp. 211-214 ◽  
Author(s):  
Roger N. F. Thorneley ◽  
Keith R. Willison

Acetylene-reducing activity of purified nitrogenase from Klebsiella pneumoniae was studied over a range of ATP and Mg2+ concentrations at 15°C, pH7.8. Inhibition at Mg2+ concentrations of 2.5–30mm was due to the formation of the inactive complex, Mg2ATP. At higher Mg2+ concentrations an additional inhibitory effect was observed. The results were consistent with a MgATP complex being the active substrate with an apparent Km(MgATP)=0.4mm.


1981 ◽  
Vol 27 (8) ◽  
pp. 815-823 ◽  
Author(s):  
John D. Tjepkema ◽  
William Ormerod ◽  
John G. Torrey

Vesicle formation and acetylene reduction (nitrogenase activity) were observed when washed hyphae from cultures of Frankia sp. CpI1 were transferred to a nitrogen-free medium containing ethylenediaminetetraacetic acid and succinate. Succinate could be replaced by malate or fumarate, but not other carbon sources. Maximum acetylene reduction and vesicle numbers were observed at a pH of 6.0–6.5, at 25–30 °C, and at atmospheric [Formula: see text] or somewhat less (5–20 kPa). Addition of 1 mM NH4Cl almost completely inhibited vesicle formation and acetylene-reducing activity, but did not immediately inhibit such reducing activity by cultures with preexisting vesicles. Acetylene-reducing activity was never observed in the absence of vesicle formation.


1992 ◽  
Vol 38 (12) ◽  
pp. 1238-1241 ◽  
Author(s):  
Anil K. Tripathi ◽  
Walter Klingmüller

Azospirillum brasilense and Azospirillum lipoferum showed optimum acetylene-reducing activity at 25 and 30 °C, respectively, although both the bacteria grew optimally at 35 °C. Azospirillum halopraeferens displayed optimum growth and acetylene-reducing activity at 40–41 °C. Our experiments indicated that expression of nif genes was generally more sensitive to temperature than was nitrogenase activity. The NifA-dependent activation of a heterologous nifH–lacZ fusion was used to assess the impact of temperature on native NifA activity of A. brasilense and A. lipoferum. Maximum NifA activity was observed at 25 °C in A. brasilense and at 30 °C in A. lipoferum. Key words: temperature, nitrogen fixation, nifH–lacZ fusion, NifA activity, Azospirillum.


1983 ◽  
Vol 61 (9) ◽  
pp. 2405-2409 ◽  
Author(s):  
F. D. H. Macdowall

Seedlings of Medicago sativa L. cv. Algonquin were grown in vermiculite and nodulated by Rhizobium meliloti strain 102F70 at two lower levels of N, until flowering when the tops were cut off to leave about 10% shoot stubble. Residual shoot dry matter immediately resumed first-order growth and maintained it throughout regrowth to second flowering. The rate constants of shoot regrowth were 34% lower (at 15 mM NO3−), 25% lower (at 1.5 mM NO3− symbiotically), or 220% higher (at zero NO3− symbiotically) than the values for 1 to 4-week-old seedlings, which indicated a radical change in physiology. Root dry matter resumed exponential growth after a 7-day recession and its recovery and yields were independent of N nutrition. The most pronounced minima occurred in the acetylene-reducing activity of nitrogenase, the kinetics of which paralleled root dry matter except that its redevelopment stopped after two-thirds of the regrowth time. The rate coefficient for the redevelopment of nitrogenase activity equalled that for its development during the seedling stage, which suggested unchanged limitations on that process until its redevelopment stopped.


1988 ◽  
Vol 20 (1) ◽  
pp. 41-61 ◽  
Author(s):  
R. P. Fritz-Sheridan ◽  
D. S. Coxson

AbstractStereocaulon virgatum Ach. has colonized lava flows deposited on the west flank of the volcano La Soufrière. The mean annual rate of acetylene reduction was 43·4 nmol C2H4 gdw−1 h−1 with maximum rates during the prevalent cloud/shroud meteorology of 101 and minimum rates during rare high insolation events of 0·63 nmol gdw−1 h−1. Percentage thallus moisture was the major variable controlling nitrogenase activity. During cloud/shroud conditions the upper 90% of the lichen canopy reduced 85% of the acetylene. Canopy shading reduced intra-canopy temperatures allowing the basal 10% of the canopy to fix nitrogen during insolation shocks. Basal portions of pseudopodetia exhibited reduced rates of ethylene production when exposed to canopy surface light intensities during cloud/shroud conditions. The recovery pattern of nitrogenase following desiccation during an insolation shock is presented. Rates of photosynthesis during cloud/shroud conditions were high, reaching 50% of those attained during saturating light intensities.


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.


1976 ◽  
Vol 22 (10) ◽  
pp. 1561-1566 ◽  
Author(s):  
George John Waughman

Acetylene reduction by the peat microflora was unaffected by light, but was sensitive to temperature, with an optimum of about 30 °C. The nitrogenase was inactivated by exposure to temperatures above about 35 °C. Activity occurred in completely anaerobic conditions; the rate of ethylene production was of the order of 0.5 nmol C2H4 ml−1h−1. Experiments with time courses indicated that exposure to oxygen caused an initial inhibition of activity followed by a period in which ethylene production was stimulated to rates much higher than in the anaerobic tests; both the inhibition and stimulation appeared to be related to the level of oxygenation. It is suggested that these results could be explained by the existence of nitrogen-fixing associations of aerobic and anaerobic bacteria in the peat. The stimulation of activity caused by exposure to air indicates that care must be taken when interpreting results of the assay obtained from waterlogged organic soils.


1988 ◽  
Vol 39 (3) ◽  
pp. 409 ◽  
Author(s):  
GN Mundy ◽  
HR Jones ◽  
WK Mason

The effect of flood irrigation on clover nitrogen fixation (acetylene reduction activity) was investigated in unfertilized pastures and pastures fertilized with 100 kg N/ha as NH4N03. During the experiment acetylene reduction activities were monitored between successive flood irrigations. The rate of ethylene production increased with decreasing soil water content, peaking 13 days after irrigation. Measured nodule activity then declined, possibly owing to water stress. Nitrogen fertilizer reduced acetylene reduction activities to about half that of the unfertilized pastures. For most of the irrigation cycle acetylene reduction by the pasture was below the measured maximum.


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