Effects of Protein Synthesis Inhibitors on Acetylene Reduction Activity of Lupin Root Nodules

1980 ◽  
Vol 7 (3) ◽  
pp. 261 ◽  
Author(s):  
WD Sutton

Rifampicin and D-threo-chloramphenicol inhibited the incorporation of [35S]methionine into purified bacteroid suspensions, and into the bacteroid fraction but not the plant cytoplasmic fraction of cultured nodules. Cycloheximide and anisomycin inhibited [35S]methionine incorporation into the plant cytoplasmic fraction of cultured nodules; at early times they inhibited incorporation into the bacteroid fraction, but at later times this effect was reversed. Chloramphenicol, rifampicin, spectinomycin, cycloheximide and anisomycin all prevented the induction of acetylene reduction activity in immature nodules; spectinomycin did not prevent induction in nodules containing a spectinomycin-resistant Rhizobium. Neither rifampicin nor chloramphenicol inhibited the acetylene reduction activity of mature nodules, but cycloheximide and anisomycin caused rapid loss of activity. Cycloheximide did not inhibit the acetylene reduction activity of Rhizobium strain 32H1 in pure cultures. The results suggest that both plant cytoplasmic protein synthesis and bacteroid protein synthesis are needed for the induction of nitrogenase activity in developing lupin nodules, and that plant cytoplasmic protein synthesis but not bacteroid protein synthesis is needed for the maintenance of nitrogenase activity at high levels.


1992 ◽  
Vol 38 (6) ◽  
pp. 577-583 ◽  
Author(s):  
J. K. Ladha ◽  
Minviluz Garcia ◽  
R. P. Pareek ◽  
G. Rarivoson

Six experiments, two each in the phytotron, greenhouse, and field, were conducted to assess the contribution of nitrogenase activity (acetylene reduction) by stem nodules in the presence and absence of root nodules of Sesbania rostrata (Brem & Oberm). In a greenhouse experiment, the effect of detaching already formed aerial stem nodules on the restoration of root nodules and nitrogenase activity was studied. The field experiment compared nodulation and acetylene-reduction activity by dual-nodulating S. rostrata and root-nodulating Sesbania cannabina. Acetylene-reduction activity expressed per gram of nodule dry weight was higher for stem nodules than for root nodules. Root nodule dry weight and acetylene-reduction activity failed to increase after stem inoculation, but root nodule dry weight and acetylene-reduction activity increased several fold within 15 days of detachment of aerial stem nodules. Stem nodulation, which occurred without inoculation under lowland field condition, suppressed root nodulation, thus accounting for more than 75% of total nitrogenase activity. Sesbania rostrata showed higher acetylene-reduction activity than S. cannabina. In dual-nodulating plants, root and stem nodules appeared to strike a balance in competition for energy, which may be controlled by stem nodulation. Key words: Sesbania rostrata, Azorhizobium caulinodans, stem nodule, root nodule, acetylene-reducing activity.



1978 ◽  
Vol 56 (18) ◽  
pp. 2218-2223 ◽  
Author(s):  
D. Smith ◽  
D. G. Patriquin

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.



1992 ◽  
Vol 72 (2) ◽  
pp. 383-390 ◽  
Author(s):  
A. Djekoun ◽  
C. Planchon

Yield limitation in soybean (Glycine max L. Merr.) can result from decreases in photosynthesis and N2 fixation during periods of water deficiency. In this study, the relationships among stomatal conductance, photosynthesis and N2 fixation were analyzed in connection with drought tolerance of genotypes. Plants were grown in pots and exposed to field conditions. Carbon dioxide exchange rate was measured by gas analysis and nodule activity by the acetylene reduction method. Leaf water status was determined with a pressure bomb, and nodule water potential and leaf osmotic potential were measured psychrometrically. The differing tolerances of the cultivars Kingsoy and Hodgson to leaf water deficit resulted in a more or less developed ability of the lower side of the leaf to maintain good stomatal conductance during water stress. Stomatal conductance affects photosynthetic rate directly and acetylene reduction activity indirectly. Early stomatal closure, by limiting H2O exchange, contributes to conservation of nitrogenase activity. On the contrary, maintenance of high conductance during a water stress decreases soil water availability and nodule water content, which in turn has a decisive and limiting effect on acetylene reduction activity. Thus, if tolerance at low leaf water potentials associated with osmotic adjustment is an important drought mechanism for maintaining photosynthetic processes under water-limited conditions, the result would be obtained at the expense of symbiotic N2 fixation.Key words: Glycine max L. Merr., nitrogenase activity, photosynthesis, drought stress, soybean



Author(s):  
C. J. Emerson ◽  
A. K. Bal

Nitrogen-fixing peanut root nodules have highly specialized, large spherical bacteroids and show higher acetylene-reduction activity than the nodules of other legumes induced by the same strain of rhizobia. Differences are also found in their anatomical organization and with regard to bacteroid-associated organelles and other structures (dense bodies) found on the host-symbiont interface. In this investigation, further description of these organelles and structures was made at different stages of development in a correlated SEM/TEM study.Arachis hypogaea L. cv. Jumbo Virginia plants were grown in environmental growth chambers and inoculated with Rhizobium sp. 32Hl. For TEM, slices of nodules were fixed in Karnovsky's fixative in Sorensen's phosphate buffer, pH 7.2 for 1 h at 4°C. Tissues were rinsed in buffer, post-fixed with 1% phosphate-buffered osmium tetroxide for 1 h at 4°C and dehydrated in ethanol. 1% p-phenylenediamine was added at the 70% ethanol step (30 min) to preserve lipids.





2005 ◽  
Vol 51 (11) ◽  
pp. 948-956 ◽  
Author(s):  
Chungwoo Kim ◽  
Mihály L Kecskés ◽  
Rosalind J Deaker ◽  
Kate Gilchrist ◽  
Peter B New ◽  
...  

Nitrogen-fixing bacteria were isolated from the rhizosphere of different crops of Korea. A total of 16 isolates were selected and characterized. Thirteen of the isolates produced characteristics similar to those of the reference strains of Azospirillum, and the remaining 3 isolates were found to be Enterobacter spp. The isolates could be categorized into 3 groups based on their ARDRA patterns, and the first 2 groups comprised Azospirillum brasilense and Azospirillum lipoferum. The acetylene reduction activity (ARA) of these isolates was determined for free cultures and in association with wheat roots. There was no correlation between pure culture and plant-associated nitrogenase activity of the different strains. The isolates that showed higher nitrogenase activities in association with wheat roots in each group were selected and sequenced. Isolates of Azospirillum brasilense CW301, Azospirillum brasilense CW903, and Azospirillum lipoferum CW1503 were selected to study colonization in association with wheat roots. We observed higher expression of β-galactosidase activity in A. brasilense strains than in A. lipoferum strains, which could be attributed to their higher population in association with wheat roots. All strains tested colonized and exhibited the strongest β-galactosidase activity at the sites of lateral roots emergence.Key words: Azospirillum, acetylene reduction activity, 16S rDNA, ARDRA patterns, lacZ fusion.



1985 ◽  
Vol 31 (9) ◽  
pp. 804-809 ◽  
Author(s):  
Marcia A. Murry ◽  
Zhang Zhongze ◽  
John G. Torrey

The effect of the partial pressure of oxygen (PO2) on the formation of vesicles, which are thought to be the site of N2 fixation in Frankia, was studied in HFPCcI3, an effective isolate from Casuarina cunninghamiana. Unlike other actinorhizal root nodules, vesicles are not produced by the endophyte in Casuarina nodules. However, in culture under aerobic conditions, large, phase-bright vesicles are formed in HFPCcI3 within 20 h following removal of [Formula: see text] from the culture medium and reach peak numbers within 72 to 96 h. In vivo acetylene reduction activity parallels vesicle formation. Optimum rates of acetylene reduction in short-term assays occurred at 20% O2 (0.2 atm (1 atm = 101.325 kPa)) in the gas phase. O2 uptake (respiration) determined polarographically showed diffusion-limited kinetics and remained unsaturated by O2 until 300 μM O2. In contrast, respiration in [Formula: see text]-grown cells was saturated by O2 between 8 and 10 μM O2. These results indicate the presence of a diffusion barrier associated with the vesicles. Vesicle development was repressed in cells incubated in N-free media sparged with gas mixtures with PO2 between 0.001 and 0.003 atm. Nitrogenase was induced under these conditions, but acetylene reduction was extremely O2 sensitive. The kinetics of O2 uptake as a function of dissolved O2 concentration in avesicular cells were similar to those in [Formula: see text]-grown cells indicating the lack of a diffusion barrier. These results demonstrate that vesicle formation and the development of the O2 protection mechanisms of nitrogenase are regulated by ambient PO2 in HFPCcI3.



1987 ◽  
Vol 14 (5) ◽  
pp. 527 ◽  
Author(s):  
AD Day ◽  
GD Price ◽  
KA Schuller ◽  
PM Gresshoff

The nodule physiology of a supernodulating, nitrate tolerant symbiosis soybean (Glycine max (L.) Merr.) mutant (nts382) was compared to that of its wild-type parent, cv. Bragg. Nodule number and mass were greater in nts382 than cv. Bragg and individual nodule mass, bacteroid and haem content, and acetylene reduction activity per g nodule were less. Acetylene reduction activity expressed per mg bacteroid protein was the same in the two genotypes. In median sections, the ratio of infected to total nodule area was smaller in nts382, infected cell size was smaller and there were fewer bacteroids per peribacteroid envelope. When inoculum dose was decreased from 109 to 103 viable cells per pot, nodule number on nts382 decreased approximately to that on cv. Bragg; nodule size, bacteroid and haem contents increased as did nodule acetylene reduction activity. Application of moderate levels of nitrate, which did not significantly affect symbiotic parameters of cv. Bragg or high inoculum nts382, stimulated nodule growth and nitrogenase activity of low inoculum nts382. A combination of nitrate and low inoculum levels enhanced nodule parameters of nts382 to the level usually seen with cv. Bragg. When supernodulated, plant dry weight of nts382 was less than that of Bragg; decreasing inoculum dose had no significant effect but nitrate application increased plant growth; nitrate plus low inoculum induced similar plant growth to that of cv. Bragg. Nodule carbohydrate content was similar in both genotypes but nodule and xylem sap ureide contents were higher in nts382. In general, nodules of supernodulated nts382 resembled under-developed cv. Bragg nodules; when supernodulation was avoided by using low inoculum doses, ,nts382 nodules resembled those of cv. Bragg. Nitrogen metabolism in the mutant seems to be disturbed, resulting in ureide accumulation.



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.



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