Acetylene reduction and hydrogen evolution by nitrogenase in a Rhizobium–legume symbiosis

1983 ◽  
Vol 61 (3) ◽  
pp. 780-785 ◽  
Author(s):  
Scott A. Edie
Keyword(s):  
Hydrogen Evolution ◽  
Acetylene Reduction ◽  
Rhizobium Leguminosarum ◽  
High Light ◽  
Hydrogenase Activity ◽  
Uptake Hydrogenase ◽  
Vegetative Phase ◽  
Pisum Sativum L ◽  
Legume Symbiosis ◽  
The Relationship

The relationship between acetylene reduction and hydrogen evolution in air was examined in peas (Pisum sativum L. cv. Alaska) inoculated with Rhizobium leguminosarum strain 3740, which lacked uptake hydrogenase activity (Hup−). In the absence of a system for recycling hydrogen, changes in the relative efficiency of N2 fixation (RE), which is defined as RE = 1 − (hydrogen evolved in air)/(acetylene reduced), presumably reflect an altered capacity of nitrogenase to allocate electrons between protons and N2. The RE of plants grown without combined nitrogen declined during the vegetative phase of growth and increased after flowering. Continuous high light or elevated CO2 conditions in the absence of nitrate maintained throughout ontogeny accentuated decreases in RE. When nitrate was present in the growth medium declines in RE during the vegetative phase were lessened. These results are consistent with the concept that the electron allocation coefficient of nitrogenase varies in the absence of uptake hydrogenase activity.

Effects of continuous combined nitrogen supply on symbiotic dinitrogen fixation of faba bean and pea inoculated with different rhizobial isolates

1987 ◽  
Vol 65 (12) ◽  
pp. 2542-2548 ◽  
Author(s):  
François-P. Chalifour ◽  
Louise M. Nelson
Keyword(s):  
Faba Bean ◽  
Acetylene Reduction ◽  
Rhizobium Leguminosarum ◽  
Combined Nitrogen ◽  
Pisum Sativum L ◽  
Legume Symbiosis ◽  
Total Plant ◽  
Continuous Presence ◽  
Symbiotic Partner ◽  
Rhizobial Isolates

Combined nitrogen (N) has adverse effects on virtually all stages of the Rhizobium–legume symbiosis. Tolerance to combined N varies among legume hosts and rhizobial isolates, but the contribution of each symbiotic partner is not well established. The effects of combined N were studied in faba bean (Vicia faba L.) and pea (Pisum sativum L.), using the same Rhizobium leguminosarum isolates for both hosts. In one experiment, faba bean and pea were inoculated individually with four rhizobial isolates and grown for 28 days in the continuous presence of 0, 2.5, 5.0, or7.5 mol m−3 NH4NO3. For each isolate, faba bean was consistently more tolerant to combined N than pea as shown by significantly smaller rates of decrease in N2-fixing activity (acetylene reduction) in faba bean than in pea. The results were substantiated by those of a similar experiment in which increasing levels of 15N-labeled [Formula: see text] (5, 10, or 15 mol m−3) were supplied continuously to faba bean and pea inoculated individually with two rhizobial isolates. Comparisons of the different symbioses based on the proportion of total plant N derived from N2 fixation confirmed the conclusions reached using acetylene reduction activities.

Pisum sativum cultivar effects on hydrogen metabolism in Rhizobium

1984 ◽  
Vol 62 (8) ◽  
pp. 1682-1686 ◽  
Author(s):  
Eulogio J. Bedmar ◽  
Donald A. Phillips
Keyword(s):  
Pisum Sativum ◽  
Relative Efficiency ◽  
Rhizobium Leguminosarum ◽  
Root Nodules ◽  
Developmental Differences ◽  
The Other ◽  
Hydrogenase Activity ◽  
Uptake Hydrogenase ◽  
Hydrogen Metabolism ◽  
Pisum Sativum L

Data from 14 Pisum sativum L. cultivars establish that three pea genotypes, which were previously reported to affect net H2 evolution from root nodules in air and uptake hydrogenase activity of Rhizobium leguminosarum 128C53, are not unique. Two pea lines, 'JI1205' and 'Green Arrow,' produced very active uptake hydrogenase activity in strain 128C53, and essentially no H2 was evolved in air from root nodules capable of reducing 20 μmol C2H2 ∙ plan−1 ∙ h−1. Five other cultivars produced significantly lower uptake hydrogenase activities in the same bacterial strain and had much higher rates of net H2 evolution with similar C2H2-reduction capabilities. Parallel experiments with the same cultivars nodulated by R. leguminosarum 300, an organism with no convincing uptake hydrogenase activity in any pea line, showed that 'JI1205' and 'Green Arrow' had a significantly lower relative efficiency (RE) of N2 fixation (1 − (H2 evolved in air/C2H2 reduced)) than the other five cultivars. Developmental differences among the pea lines prevented any conclusion about the advantages or disadvantages of uptake hydrogenase activity for plant growth, but in general, cultivars with high uptake hydrogenase activity and low net H2 evolution grew more slowly than those evolving large amounts of H2.

scholarly journals Generation of New Hydrogen-RecyclingRhizobiaceae Strains by Introduction of a Novelhup Minitransposon

2000 ◽  
Vol 66 (10) ◽  
pp. 4292-4299 ◽  
Author(s):  
Elena Báscones ◽  
Juan Imperial ◽  
Tomás Ruiz-Argüeso ◽  
Jose Manuel Palacios
Keyword(s):  
Sinorhizobium Meliloti ◽  
Rhizobium Leguminosarum ◽  
General Procedure ◽  
Large Subunit ◽  
Antibiotic Resistance Gene ◽  
Insertion Site ◽  
Immunoblot Analysis ◽  
Hydrogenase Activity ◽  
Mesorhizobium Loti ◽  
Legume Symbiosis

ABSTRACT Hydrogen evolution by nitrogenase is a source of inefficiency for the nitrogen fixation process by the Rhizobium-legume symbiosis. To develop a strategy to generate rhizobial strains with H2-recycling ability, we have constructed a Tn5derivative minitransposon (TnHB100) that contains the ca. 18-kb H2 uptake (hup) gene cluster fromRhizobium leguminosarum bv. viciae UPM791. Bacteroids from TnHB100-containing strains of R. leguminosarum bv. viciae PRE, Bradyrhizobium japonicum, R. etli, and Mesorhizobium loti expressed high levels of hydrogenase activity that resulted in full recycling of the hydrogen evolved by nitrogenase in nodules. Efficient processing of the hydrogenase large subunit (HupL) in these strains was shown by immunoblot analysis of bacteroid extracts. In contrast, Sinorhizobium meliloti,M. ciceri, and R. leguminosarum bv. viciae UML2 strains showed poor expression of the hup system that resulted in H2-evolving nodules. For the latter group of strains, no immunoreactive material was detected in bacteroid extracts using anti-HupL antiserum, suggesting a low level of transcription ofhup genes or HupL instability. A general procedure for the characterization of the minitransposon insertion site and removal of antibiotic resistance gene included in TnHB100 has been developed and used to generate engineered strains suitable for field release.

scholarly journals Development and Partial Characterization of Nearly Isogenic Pea Lines (Pisum sativum L.) that Alter Uptake Hydrogenase Activity in Symbiotic Rhizobium

1990 ◽  
Vol 92 (4) ◽  
pp. 983-989 ◽  
Author(s):  
Donald A. Phillips ◽  
Yoram Kapulnik ◽  
Eulogio J. Bedmar ◽  
Cecillia M. Joseph
Keyword(s):  
Pisum Sativum ◽  
Partial Characterization ◽  
Hydrogenase Activity ◽  
Uptake Hydrogenase ◽  
Pisum Sativum L

scholarly journals Uptake Hydrogenase Activity Determined by Plasmid pRL6JI in Rhizobium leguminosarum Does Not Increase Symbiotic Nitrogen Fixation

1985 ◽  
Vol 50 (4) ◽  
pp. 791-794 ◽  
Author(s):  
Scott D. Cunningham ◽  
Yoram Kapulnik ◽  
Nicholas J. Brewin ◽  
Donald A. Phillips
Keyword(s):  
Nitrogen Fixation ◽  
Rhizobium Leguminosarum ◽  
Symbiotic Nitrogen Fixation ◽  
Hydrogenase Activity ◽  
Uptake Hydrogenase

Nickel-Dependent Uptake-Hydrogenase Activity in the Blue-Green Alga Anabaena variabilis

1984 ◽  
Vol 39 (1-2) ◽  
pp. 90-92 ◽  
Author(s):  
Helmar Almon ◽  
Peter Böger
Keyword(s):  
Hydrogen Evolution ◽  
Green Alga ◽  
Nickel Chloride ◽  
Anabaena Variabilis ◽  
Hydrogenase Activity ◽  
Uptake Hydrogenase ◽  
Blue Green Alga ◽  
Hydrogen Formation ◽  
Permeabilized Cells ◽  
Cyanobacterium Anabaena

Abtract Uptake-hydrogenase activity in the blue-green alga (cyanobacterium) Anabaena variabilis is dependent on the presence of nickel. Nickel depletion leads to an enhancement of net lightinduced hydrogen evolution, which is catalyzed by nitrogenase. Addition of nickel chloride to the culture reverses this effect by stimulating hydrogenase activity. Methylviologen/dithionitedriven hydrogen formation of permeabilized cells is decreased by nickel depletion.

scholarly journals Uptake hydrogenase activity and ATP formation in Rhizobium leguminosarum bacteroids.

1982 ◽  
Vol 151 (2) ◽  
pp. 989-995 ◽  
Author(s):  
L M Nelson ◽  
S O Salminen
Keyword(s):  
Rhizobium Leguminosarum ◽  
Hydrogenase Activity ◽  
Uptake Hydrogenase ◽  
Atp Formation

Nitrogen fixation and hydrogen metabolism by Rhizobium leguminosarum isolates in pea root nodules

1981 ◽  
Vol 27 (10) ◽  
pp. 1028-1034 ◽  
Author(s):  
Louise M. Nelson ◽  
J. J. Child
Keyword(s):  
Relative Efficiency ◽  
Rhizobium Leguminosarum ◽  
High Efficiency ◽  
Root Nodules ◽  
Dry Weight ◽  
Hydrogenase Activity ◽  
Uptake Hydrogenase ◽  
Hydrogen Metabolism ◽  
N Content ◽  
Significant Uptake

A survey of 108 isolates of Rhizobium leguminosarum was conducted to determine the variation in H2 uptake and relative efficiency of N2 fixation in Pisum sativum L. root nodules and the relation of relative efficiency to plant dry weight and N content. Only 14 of the isolates exhibited significant uptake hydrogenase activity and none of these had sufficient hydrogenase activity to recycle all of the H2 produced by nitrogenase. In 74 of the isolates tested relative efficiencies of N2 fixation were less than 0.60.Twenty-nine of the isolates were ineffective, since total plant N at harvest did not differ significantly from uninoculated controls. The remaining 79 effective isolates could be divided into low-and high-efficiency groups. Plants which were inoculated with isolates from the two groups and harvested after 4 weeks did not differ significantly in plant dry weight or N content. Among the isolates with high relative efficiency of N2 fixation, two groups could be recognized: one possessing significant uptake hydrogenase activity, the other lacking hydrogenase activity but in which H2 evolution was low. Although the two groups did not differ with respect to plant dry weight or N content, the identification of this latter group may be of some significance for optimizing the efficiency of N2 fixation.

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