The role of oxygen diffusion from the shoots and nodule roots in nitrogen fixation by root nodules of Myrica gale

1978 ◽  
Vol 56 (11) ◽  
pp. 1365-1371 ◽  
Author(s):  
John Tjepkema

Nitrogenase activity (acetylene reduction) and oxygen uptake by root nodules of Myrica gale L. were measured before and after removal of nodule roots. There was no significant effect of nodule root removal when respiration was measured in the gas phase (0.05–0.2 atm pO2 (1 atm = 101.325 kPa)) or acetylene reduction in a stirred aqueous phase at 0.2 atm pO2. However, when acetylene reduction was measured in 0.05 atm pO2 in an unstirred aqueous phase, there was a 66 to 76% reduction in activity. These results indicate that nodule roots are important for oxygen uptake when the nodules are present in an aqueous phase at low pO2, which is probably the normal environmental conditions for many of the nodules. Other measurements showed that diffusion of oxygen from the shoot to the root nodules is not important for nitrogen fixation. These measurements were done on whole plants with the shoots in air (0.20 atm pO2) and the roots in water at the desired pO2 value. With 0.0 atmpO2 in the root environment, the rate of acetylene reduction was only 4% of the rate at 0.2 atmpO2. Thus, only small amounts of oxygen are transported from the shoot to the nodules.


2001 ◽  
Vol 79 (9) ◽  
pp. 1010-1018 ◽  
Author(s):  
Christa R Schwintzer ◽  
John D Tjepkema

In wet soils, nitrogen-fixing root nodules are subjected to elevated CO2. Only a few studies have examined the effect of elevated CO2 on nitrogenase activity, and the results have been mixed. We examined intact black alder (Alnus glutinosa (L.) Gaertn.) and sweet gale (Myrica gale L.) seedlings and field-collected speckled alder (Alnus incana ssp. rugosa (Du Roi) Claus.) nodules to clarify the effects of elevated CO2. Nitrogenase activity was measured via acetylene reduction in an open, flow-through system. We found that repeated measurements of the peak rate of nitrogenase activity, the only reliable measure of nitrogenase activity, could be made on the same plant via 150-s exposures to acetylene separated by 20 min without acetylene. Our results for elevated CO2 consistently showed that it had little effect on nitrogenase activity at low concentrations and increasingly inhibited nitrogenase activity as the CO2 concentration increased. In black alder, 0.5 kPa CO2 had little effect, whereas 3.0 kPa CO2 reduced nitrogenase activity 31–35%. Sweet gale, was less sensitive to elevated CO2 and was unaffected by 1.5 kPa CO2. Black alder grown with the roots, but not the shoots, in 1.3 kPa CO2 showed only minimal acclimation to elevated CO2.Key words: acetylene reduction technique, actinorhizal plants, Alnus, carbon dioxide, Myrica gale, nitrogen fixation.



1981 ◽  
Vol 29 (2) ◽  
pp. 143 ◽  
Author(s):  
AC Lawrie

Nitrogen fixation was studied by the acetylene-reduction technique in root nodules of 10 native Australian legumes in three habitats in Victoria: low open-forest. sandy heathland and coastal sand dunes. Nodular activity reached a maximum in spring (and in autumn in sand-dune species), declined to a minimum in late summer and continued at a low level throughout winter. Mean nitrogenase activity was 1.73 μmol C2H2 reduced g-1 fresh wt nodules h-1, with spring maxima 8-250 times summer minima. It is suggested that variations in rainfall and temperature were the major factors influencing seasonal variations in nodular activity. Estiniates of minimum acetylene reduction per hectare per year were derived from measurements of seasonal and diurnal variations in acetylene reduction and abundance of nodules per hectare, and were used to derive approximate minimum values for nitrogen fixation of 0.004-0.746 kg ha-1 yr-1. The quantities of nitrogen fixed are compared with other reports and the significance of nitrogen fixation by these native legumes is discussed.



1986 ◽  
Vol 32 (4) ◽  
pp. 353-358 ◽  
Author(s):  
Mary F. Lopez ◽  
Patricia Young ◽  
John G. Torrey

The carbon source requirements for the growth and nitrogen fixation of two morphologically distinct Frankia isolates were examined. Isolate ArI3 (from Alnus rubra) grew well on propionate, malate, acetate, and trehalose, and isolate CcI2 (from Casuarina cunninghamiana) grew best on pyruvate, acetate, and propionate. In general, the same carbon sources that supported growth supported both the development of vesicles and nitrogenase activity in long-term induction experiments in both isolates. However, ArI3 cultures induced on proprionate had 7 to 26 times the activity of other carbon sources and ArI3 cultures induced on acetate did not develop any detectable acetylene reduction. In a parallel set of experiments, cultures of both isolates were induced for nitrogenase activity on propionate and the resulting nitrogen fixing cultures were washed free of the organic acid by centrifugation. The washed cultures were incubated in the presence of various carbon sources to determine the ability of a particular substrate to supply energy directly for nitrogen fixation when vesicles and nitrogenase were already present. As was observed in the long-term induction experiments, pyruvate, propionate, and acetate supported the greatest activity in CcI2. Succinate and malate supported the greatest activity in ArI3, and propionate had very little stimulation of acetylene reduction. The reason for the lack of stimulation by propionate for washed cells of ArI3 was unclear but may have been due to toxic concentrations of the organic acid. In an attempt to compare the carbon utilization of ArI3 in pure culture with that in the alder symbiosis, oxygen uptake in the presence of various carbon sources of vesicles clusters isolate from Alnus rubra nodules inoculated with ArI3 was compared with the oxygen uptake of nitrogen-fixing pure cultures of ArI3. The oxygen uptake of the isolated vesicle clusters was stimulated by sucrose, trehalose, and glucose, but not by a variety of organic acids. In comparison, nitrogen-fixing pure cultures of ArI3 readily oxidized sugars and organic acids.





1983 ◽  
Vol 61 (11) ◽  
pp. 2937-2942 ◽  
Author(s):  
Christa R. Schwintzer ◽  
John D. Tjepkema

Annual CO2 evolution, H2 evolution, and C2H2 reduction were measured in root nodules from a vigorous Myrica gale stand in a Massachusetts peatland at 3-week intervals in 1980. Nodule activity was approximately the same under the experimental conditions (excised nodules reducing C2H2) as in nature (attached nodules reducing N2) and the CO2 evolution to O2 uptake ratio averaged 1.07. Nitrogenase activity was first detectable in late May, reached its maximum [Formula: see text] in mid-July, and disappeared in late October. The seasonal pattern of CO2 evolution was similar except that it continued at low rates when nitrogenase activity was absent. Hydrogen evolution was barely detectable. The energy cost of nitrogen fixation, expressed as the molar CO2:C2H4 ratio, was relatively low [Formula: see text] throughout the period of substantial nitrogenase activity and had a mean annual value of 4.9. Annual N2 fixation was estimated to be 2.8 g N m−2year−1, contributing about 33% of the annual N requirement measured in 1979. Annual C use by nodules was about 21.0 g C m−2 year−1. If this C were available for additional net production, it would increase it by about 5.5%.





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.



1983 ◽  
Vol 10 (1) ◽  
pp. 26-29 ◽  
Author(s):  
P. T. C. Nambiar ◽  
P. J. Dart

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.



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.



1969 ◽  
Vol 15 (7) ◽  
pp. 671-675 ◽  
Author(s):  
G. S. Hendry ◽  
D. C. Jordan

Under clearly defined conditions one-step acquisition of viomycin resistance by a normally effective strain of Rhizobium meliloti resulted in one-step acquisition of ineffectiveness in nitrogen fixation, which probably occurred with a one-gene change in the R. meliloti genome. Two-step mutants retained their ability to produce root nodules but such nodules also were ineffective. Increased sensitivity of the viomycin-resistant mutants to glycine and D-alanine was not noted. Bacteroids were not seen in nodules formed by the viomycin-resistant mutants on their homologous host plant. Nitrogenase activity was not detected, by acetylene reduction, in detached ineffective nodules, whereas effective nodules formed 10.6 μmoles of ethylene per hour per gram of nodules. Growth of the effective parent strain in a low concentration of viomycin resulted in elongation and swelling of the cells so that they appeared as artificially produced bacteroids. Viomycin-resistant mutants did not undergo this transformation. Antigens could be readily extracted by hot- and cold-saline extraction of wet packed cells of both resistant and sensitive cultures but antigenic differences, which may have indicated cell wall differences, were not noted.



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