Temperature sensitivity of nitrogen fixation in Azospirillum spp.

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.


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.



1992 ◽  
Vol 38 (8) ◽  
pp. 794-797 ◽  
Author(s):  
K. M. Khammas ◽  
P. Kaiser

Cocultures of different Azospirillum species with Bacillus polymyxa or Bacillus subtilis allow the efficient utilization of pectin as carbon and energy sources for nitrogen fixation. The nitrogenase activity obtained with cocultures was as high as 30–80 nmol C2H4 h−1 mL−1, a much higher value than that obtained with pure cultures of either Azospirillum (up to 13 nmol C2H4 h−1 mL−1) or B. polymyxa (up to 2 nmol C2H4 h−1 mL−1) alone. To establish to what extent each partner contributed to nitrogenase activity, acetylene reduction was assayed as a function of time and it was also measured on Azospirillum cultivated in the cultures filtrates of the Bacillus. The results suggested that the nitrogenase activity was mostly produced by Azospirillum. The nitrogenase activity occurred at the expense of the degradation and fermentation products of the pectin. The new pectinolytic species, Azospirillum irakense, utilized both degradation and fermentation products of pectin, whereas the nonpectinolytic strains (Azospirillum brasilense, Azospirillum lipoferum, Azospirillum amazonense) utilized only the fermentation products of pectin, including acetic and succinic acids. These cocultures can be considered as metabolic associations, where the Bacillus produces degradation and fermentation products of pectin, which can be used by Azospirillum species. Key words: cocultures, nitrogen fixation, pectin degradation, Azospirillum, Bacillus, metabolic association.



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.



1985 ◽  
Vol 104 (1) ◽  
pp. 207-215 ◽  
Author(s):  
R. Rai

SummaryNitrosoguanidine-induced mutation frequencies for resistance to streptomycin, spectinomycin, erythromycin and novomycin were studied inAzospirillum brasilense.Lentil inoculated withA. brasilenseand its mutants andRhizobiumstrains produced increased nodule dry weight, nitrogenase activity of nodules and roots and grain yield compared with an uninoculated control.



1986 ◽  
Vol 32 (4) ◽  
pp. 285-288 ◽  
Author(s):  
Devender K. Jain ◽  
R. J. Rennie

The spermosphere model showed very high seedling-to-seedling variation in the ability to induce acetylene-reducing activity in bacteria associated with wheat roots. Several seedlings failed to induce nitrogenase in N2-flxing strains of Bacillus and Azospirillum brasilense. Increasing the concentration of yeast extract or the inoculum density, adding NaOH solution in the side arm of Pankhurst tubes to absorb CO2, and vernalizing the seeds did not increase the percentage of nitrogenase-positive tubes and did not decrease variation in acetylene-reducing activity. Increasing the incubation temperature from 25 to 27 °C induced nitrogenase in Bacillus, but not in Azospirillum strains; at 30 °C nitrogenase was also induced in Azospirillum strains. It appears that the spermosphere model is not a practical technique for screening nitrogenase induction by wheat, unless the use of genetic lines of unimpeachable uniformity can provide control of variability.



HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1061A-1061
Author(s):  
Laura M. R. Rinaldi ◽  
Maria C. Margheri ◽  
Alba Ena

Cycasrevoluta Thunb., cultivated as an ornamental plant for indoor and outdoor use, is characterized by an extremely slow rate of growth. In spite of the occurrence in its coralloid roots of the nitrogen-fixing cyanobiont Nostoc, N fertilization is commonly used to accelerate Cycas growth. A greenhouse experiment was conducted to examine the effects of two forms of combined N on growth of Cycas plants and cyanobacterial nitrogenase activity, measured on intact plants. Cycas plants grown in pots were fertilized from June to September with nutrient solution containing macronutrients as P, K, Mg, and Hoagland's micronutrients. N (700 mg/plant) was supplied as KNO3, or applied as NH4NO3; control plants received nutrient solution without nitrogen. Treatments were applied monthly and repeated for three times. Each treatment consisted of 15 plants. The length of the new leaves was recorded during the growth cycle of the plants. The nitrogenase activity, based on acetylene reducing activity (ARA), was measured on the plants in situ in July and in October. N fertilization stimulated both the nitrogenase activity and the growth of the plants. In comparison with the control, average increases in ARA of more than 20% were observed in the treated plants. Nitrogenase activity was slightly better in the presence of NH4NO3 in July, whereas the values measured in October were about the same for two treatments. The two forms of nitrogen were the same also regarding the stimulus on growth: in N-treated plants the total length of the new leaves was more than double with respect to the control at the end of the growing season. Control plants grown without fertilizer N had a slow start to their growth cycles and were unable to recover and compensate later for the lack of transient N.



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.



1976 ◽  
Vol 22 (4) ◽  
pp. 523-530 ◽  
Author(s):  
Arthur D. Nelson ◽  
Lynn E. Barber ◽  
John Tjepkema ◽  
Sterling A. Russell ◽  
Robert Powelson ◽  
...  

Nitrogen fixation associated with both natural grasslands and grain crops of Oregon was studied using the acetylene-reduction assay. A number of the grasses collected had some acetylene-reducing activity. Agrostis tenuis Sibth. had substantially greater activity than any of the other species, with a mean rate estimated at 37 g N2 fixed per hectare per day. Assuming 100 days of activity, about 3 kg of N2 would be fixed per hectare per year. This quantity of nitrogen may be important in the maintenance of this species under natural conditions. Nitrogen-fixing microorganisms were isolated from the root surfaces of some of the grasses. Cultures of Bacillus macerans, Bacillus polymyxa, and Enterobacter cloacae were isolated from wheat roots as were two cultures which have not been assigned a specific taxonomic classification. Strains of N2-fixing Bacillus species and Gram-negative aerobic bacteria were isolated from Festuca and Agrostis.



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