Nitrogenase activity during chinook snowmelt sequences by Nostoc commune in Stipa–Bouteloa grassland

1983 ◽  
Vol 29 (8) ◽  
pp. 938-944 ◽  
Author(s):  
D. S. Coxson ◽  
K. A. Kershaw

Inactivation of the nitrogenase enzyme in winter has been observed for many terrestrial cyanophytes of north temperate habitats. In southern Alberta frequent winter chinook activity causes repeated snowmelt sequences. The pattern of nitrogenase activity in these periods has been examined (acetylene-reduction technique) over several snowmelt periods. Nostoc collected from snowmelt pockets was immediately assayed for nitrogenase activity under laboratory (20 °C, 200 E∙m−2s−1 photosynthetically active radiation (PAR)) and field (ambient light and temperature) conditions. Air and thallus temperatures for a typical snowmelt sequence rose from −20 to +5 and −5 to +20 °C, respectively, as chinook warming progressed. Activity on initial snowmelt emergence was 420 nmol C2H4∙g−1∙h−1, rising to 1050 nmol after 7 h, monitored in lab conditions. Field activity reached 90 nmol after 4 h, ceasing at 1800 as temperatures fell below 0 °C. Laboratory rates at morning thaw, day 2, were 1040 nmol, while field activity (3 °C, 900 μE) was 800 nmol. By midday laboratory rates reached 1198 nmol, while field rates (10 °C, 1500 μE) were 1077 nmol. Although these rates are well below those at the temperature optimum of 35 °C (6805 nmol), clearly no winter inactivation of the nitrogenase enzyme occurs here. Cumulative activity over winter chinook sequences may well exceed that occurring during infrequent summer hydration.

1983 ◽  
Vol 61 (10) ◽  
pp. 2686-2693 ◽  
Author(s):  
D. S. Coxson ◽  
K. A. Kershaw

Field rates of nitrogenase activity by N. commune were followed over a 1-month summer period in Stipa–Bouteloa grassland of southern Alberta, using repeated short-term acetylene-reduction assays. Concurrent patterns of temperature, moisture, and light at the soil surface were continuously monitored. There is a rapid response of nitrogenase activity to both small precipitation events and dewfall hydration periods. Typically, thalli wet by thundershower activity in the previous evening showed a midday maximum of acetylene reduction of up to 5 nmol C2H4 mg−1 h−1, followed by a rapid cessation of activity as thalli desiccated below critical moisture levels. Over extended precipitation periods rates continue to climb throughout the day, gradually falling over the night period. Dry thalli frequently experience surface temperatures of up to 65 °C, without any discernable effects on rehydration recovery of nitrogenase activity. Equally, ground frosts early in the study period did not affect subsequent activity. Surface densities of up to 23.9 g m−2 were found for N. commune, with a mean cover of 4.3 g m−2. Mean cumulative nitrogen fixation over the 31-day field period was estimated to be 10.0 mg N m−2, although up to 77.3 mg N m−2 might be expected in localized microsites. The utilization of small rainfall events in large part accounts for the magnitude of observed nitrogenase activity. Although subsequent losses of fixed nitrogen through denitrification and volatilization are yet unknown, a significant proportion of nitrogen input to this grassland site must originate from asymbiotic biological fixation.


1983 ◽  
Vol 61 (10) ◽  
pp. 2658-2668 ◽  
Author(s):  
D. S. Coxson ◽  
K. A. Kershaw

The effects of wetting–drying cycles on patterns of carbon dioxide exchange and acetylene reduction were examined for colonies of the terrestrial cyanophyte Nostoc commune collected from a semiarid grassland site in southern Alberta, Canada. At 14 and 21 °C acetylene reduction takes ca. 14 h to resume maximal rates, although activity is detected within minutes of rehydration. Net photosynthesis reaches compensation minutes after rehydration and is maximal in under 80 min at 21 °C. No respiratory burst is evident, either for replicates previously dried rapidly (0.5 h) or slowly (5 h). In marked contrast, however, at 7 °C only minimal recovery of acetylene reduction is evident after 48 h diurnal treatment, while net photosynthesis requires a recovery time of 6 h. Again no respiratory burst is evident. After rehydration, the magnitude of acetylene reduction is strongly dependent on both the previous illumination level and temperature. The rapid decline in activity on transfer to darkness or on addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea can be moderated by increasing the duration of the previous light exposure or by incubation at lower temperature. This suggests the presence of substrate pools, the replenishment of which by photosynthesis allows oxidative phosphorylative support of nitrogenase activity in the dark. The extreme resistance of N. commune to desiccation and heat stress combined with its ability to resume metabolic activity within minutes of wetting should allow very effective utilization of small precipitation events during the summer months. The slower recovery of nitrogenase activity upon rehydration at lower temperatures may restrict winter field activity on rare occasions when thalli become dehydrated between snowfall periods.


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.


1990 ◽  
Vol 17 (5) ◽  
pp. 489 ◽  
Author(s):  
Herdina ◽  
JH Silsbury

Methods of conducting acetylene reduction (AR) assay were appraised for estimating the nitrogenase activity of nodules of faba bean (Vicia faba L.). Factors considered were: (i) disturbance of plants when removing the rooting medium; (ii) assay temperature; (iii) the use of whole plants rather than detached, nodulated roots; (iv) diurnal variation in nodule activity; and (v) a decline in C2H4 production after exposure to C2H2. Plants growing in jars of 'oil dry' (calcined clay) had the same AR activity when assayed in situ in a closed system as when assayed after removal of the rooting medium. Assay temperatures of 12.5, 17.5 and 22.5°C influenced the specific rate of AR with the optimum at 17.5°C. Removal of the shoot resulted in a rapid decrease in AR activity in both vegetative and reproductive plants but the effect was much larger in the latter. AR and respiration by nodulated roots were closely linked and both varied markedly over a diurnal 12 h/12 h cycle. Since no fluctuation was found after nodules were detached, diurnal variation in the respiration of nodulated roots is attributed to change in nodule activity. Half of the dark respiration of nodulated roots was associated with respiration of the nodules and thus largely with N2 fixation. Since the AR assay provides no information on how electron flow in vivo is partitioned between reduction of N2 and reduction of protons, diurnal variation in hydrogen evolution (HE) in air and Ar/O2 in an open system was used to estimate this partitioning. Diurnal variation in apparent N2 fixation estimated in this manner was examined at a 'low' PPFD (300 μmol m-2 s-1) and at 'high' (1300 μmol m-2 s-1) to explore whether variation could be attributed to change in carbohydrate supply. Although HE in air and in Ar/O2 were both closely linked with the respiration of the nodulated root, apparent N2 fixation showed only a slight diurnal variation at 'low' light and almost none at 'high'. Vegetative plants showed no C2H2-induced decline in activity with exposure to C2H2 but reproductive plants did. This difference appears to be an age effect rather than attributable to flowering per se, since a decline occurred even when plants were kept vegetative by disbudding. A closed system for AR assay appears satisfactory for vegetative faba bean but such an assay over a 40-min period during the reproductive stage would underestimate nitrogenase activity by about 20%.


1983 ◽  
Vol 61 (11) ◽  
pp. 2956-2963 ◽  
Author(s):  
U. Granhall ◽  
T. Ericsson ◽  
M. Clarholm

The effects of single large or repeated, exponentially increasing applications of nutrients, with or without inorganic nitrogen and at two pH levels, on the growth, nodulation, acetylene reduction, and nutrient uptake in Alnus incana (L.) Moench were investigated in pot experiments with peat under controlled laboratory conditions. The repeated application of inorganic nitrogen did not suppress nitrogenase activity until the last 2 weeks, whereas an initial, large, nitrogen application effectively inhibited nodulation and activity throughout the 40-day experimental period. The mode of nitrogen application was thus found to be more important than the total amounts applied. Shoot length, leaf area, shoot–root relations, dry-matter production, and nitrogen contents of plants were determined at the end of the experiment, as well as the effect of Frankia inoculations. Nitrogenase activity was determined three times, at 0, 3, and 5 weeks. N2 fixation (balance/acetylene reduction) was found to be maximal, 55% of total nitrogen uptake, in minus-N pots with single applications of essential nutrients. The fastest growth was, however, noted in pots with single applications of all nutrients, including N. Among the latter, pots inoculated with Frankia showed the best growth, in spite of low nitrogenase activity. The only noticeable effect of a raised pH level was a reduced endophyte activity in minus-N pots with single applications of essential nutrients, due to increased N mineralization in the peat.


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.


1994 ◽  
Vol 72 (7) ◽  
pp. 940-945 ◽  
Author(s):  
R. Lennihan ◽  
D. M. Chapin ◽  
L. G. Dickson

Nostoc commune, a colonial cyanobacterium, has been suggested as an important contributor of nitrogen to terrestrial ecosystems in the Canadian High Arctic, yet little is known about the ecophysiology of this organism in arctic environments. This study focused on the physiological performance of macroscopic colonies of N. commune found on Devon Island, N.W.T. The objectives were to examine the influence of temperature, colony morphology, and seasonal phenology on nitrogen fixation rates and the effects of light and temperature on photosynthesis. Maximum rates of acetylene reduction in N. commune (2119 nmol C2H4∙g−1∙h−1) were higher than those previously recorded for arctic N. commune but lower than values reported for temperate poulations. Depending on the time of the growing season, the temperature optimum for acetylene reduction varied from 15 °C to greater than 20 °C. Photosynthetic temperature optima did not occur below 20–25 °C (the highest temperatures measured). Light saturation of photosynthesis was reached at low levels of irradiance (100–150 μmol∙m−2∙s−1 PPFD). Acetylene reduction rates varied strongly with colony morphology. Thin, fragile, flattened colonies had higher rates than thicker, more resilient, flattened colonies or spherical colonies. Cold post-thaw temperatures appeared to delay the recovery of maximum nitrogen fixation rates for 2–3 weeks following the onset of the growing season. Compared with two other species of cyanobacteria present on Truelove Lowland (Gloeocapsa alpina and Gleotrichia sp.), N. commune had higher rates of nitrogen fixation. Key words: Nostoc commune, cyanobacteria, High Arctic, nitrogen fixation, photosynthesis.


2016 ◽  
Vol 8 (1) ◽  
pp. 284-289
Author(s):  
S. Kanimoli ◽  
K. Kumar

The present study was carried out to evaluate the nitrogen fixing ability of diazotrophs isolated from the rhizosphere soils of rice which were grown in three different rice growing systems. A total of hundred and ten isolates obtained were subjected to Acetylene Reduction Assay (ARA) and ninety eight isolates recorded significant amount of nitrogenase activity in a range of 185.73 to 3794.55 nmoles of ethylene mg of protein-1 h-1. The highest nitrogenase activity was recorded by Derxia (3794.55 nmoles of ethylene mg of protein-1 h-1) isolated from Trichy (lowland). Among the three different rice production systems, isolates obtained from lowland rice (Derxia – 3794.5 nmoles of ethylene mg of protein-1 h-1) recorded higher nitrogenase activity followed by Aerobic (Pseudomonas - 2194.89 nmoles of ethylene mg of protein-1 h-1) and SRI (Azotobacter - 1971.85 nmoles of ethylene mg of protein-1 h-1) rice isolates. The results revealed marked variation in the ARA of the diazotrophic isolates obtained from lowland, SRI and Aerobic rice. The nitrogenase activity of diazotrophs from rice fields have been reported earlier but the nitrogenase activity of diazotrophs from three different rice production systems from various parts of Tamil Nadu is reported for the first time from India.


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