Nitrogen Fixation Associated with Rinsed Roots and Rhizomes of the Eelgrass Zostera marina

Nitrogen fixation was associated with the rinsed roots and rhizomes of the seagrass, Zostera marina L. Nitrogenase activity (acetylene reduction) was greater on rhizomes compared to roots, and on older roots and rhizomes relative to younger tissue. Compared to aerobic assays, anaerobic or microaerobic conditions enhanced the rate of acetylene reduction by rhizomes with attached roots, with the highest activity (100 nanomoles per gram dry weight per hour) occurring at pO₂ = 0.01 atmosphere. Addition of glucose, sucrose, or succinate also increased the rate of acetylene reduction under anaerobic conditions, with glucose providing the most stimulation. In one experiment, comparison of acetylene reduction assays with ¹⁵N₂ incorporation yielded a ratio of about 2.6:1. Seagrass communities are thought to be limited by the availability of nitrogen and, therefore, nitrogenase activity directly associated with their roots and rhizomes suggests the possibility of a N₂-fixing flora which may subsidize their nutritional demand for nitrogen.     

Nitrogen translocation via rhizome systems in monoclonal stands ofReynoutria japonica in an oligotrophic desert on Mt Fuji: Field experiments

Mechanisms which enableReynoutria japonica, a dominant pioneer herb, to be successful in maintaining large stands in an oligotrophic volcanic desert on Mt Fuji were investigated with special reference to its nitrogen acquisition.Reynoutria japonica forms circular stands, each of which comprises only one genet. As a stand develops outwards, the number of aerial shoots per unit area decreases in the center. Shoots grow vigorously in the peripheral area where the available nitrogen from soil and precipitation (about 2.4 g m⁻² year⁻²) was much less than total nitrogen in the shoots (6.1–9.1 g m⁻²). Leaf nitrogen content per unit mass was also greater in the leaves of the peripheral shoots. When rhizomes extending radially from the center to the periphery were severed, the dry mass of shoots in the periphery diminished by 75% on a ground area basis. In the periphery, leaf nitrogen content also reduced significantly and no flowers were produced. When fertilizer was applied to the peripheral shoots with severed rhizomes, neither growth, survival nor flower production of the shoots was significantly smaller than the control levels. In these shoots, it is also found that the nitrogen content in the youngest leaves decreased for about 1 month and then increased to above that in the control leaves. These results suggest that (i) nitrogen accumulated in the central part is translocated to peripheral shoots via rhizomes, and that long-distance translocation enables the stands to develop outwards, and (ii) aerial shoots in the periphery utilize the nitrogen translocated by rhizomes in the beginning of the growth season, whereas once the shoots have established, they begin to take up nitrogen with their own roots. Since the peripheral shoots are in sunnier environments than the shoots inside the stand, the acropetal nitrogen translocation via rhizomes will raise the production efficiency of a whole stand.     

Nitrogen‐dependent bacterial community shifts in root,rhizome and rhizosphere of nutrient‐efficient Miscanthus x giganteus from long‐term field trials

The perennial energy crop Miscanthus × giganteus is recognized for its extraordinary nitrogen‐use efficiency. While the remobilization of nitrogen (N) to the rhizome after the growth phase contributes to this efficiency, the plant‐associated microbiome might also contribute, as N‐fixing bacterial species had been isolated from this grass. Here, we studied established Miscanthus × giganteus plots in southern Germany that either received 80 kg N ha^?1 a^?1 or that were not N‐fertilized for 14 years. The bacterial communities of the bulk soil, rhizosphere, roots and rhizomes were analysed. Major differences were encountered between plant‐associated fractions. Nitrogen had little effect on soil communities. The roots and rhizomes showed less microbial diversity than soil fractions. In these compartments, Actinobacteria and N‐fixing symbiosis‐associated Proteobacteria depended on N. Intriguingly, N₂‐fixing‐related bacterial families were enriched in the rhizomes in long‐term zero N plots, while denitrifier‐related families were depleted. These findings point to the rhizome as a potentially interesting plant organ for N fixation and demonstrate long‐term differences in the organ‐specific bacterial communities associated with different N supply, which are mainly shaped by the plant.     

The uptake and distribution of 15N2 into the various organs of Typha latifolia L.

Direct evidence of heterotrophic dinitrogen fixation associated with the emergent aquatic angiosperm, Typha latifolia L., was obtained through the exposure of actively growing plants to ¹⁵N₂ gas for 7 days in a gas-tight exposure vessel. Highest enrichments of ¹⁵N were found in roots/rhizomes and leaf bases. Slight enrichments were also found in the leaves due to translocation from the roots, rhizomes and leaf bases. Total fixed ¹⁵N values were 71.8 μg for the plant and 49.1 μg for the soil.Plants growing in silica sand, which received a nutrient solution containing combined nitrogen, exhibited higher enrichments and fixed 86% more ¹⁵N after exposure to ¹⁵N₂ gas than plants which received a nutrient solution lacking combined nitrogen. It is hypothesized that the concentration of combined nitrogen added was insufficient to repress nitrogen fixation and resulted in an increase in nitrogen fixation by associated microorganisms.Propane was used to trace the loss and movement of gases from the ¹⁵N₂ vessel and between the upper leaf chamber and the lower root chamber. Gas was rapidly exchanged between the upper and lower chambers through the leaves and roots of T. latifolia. Further investigations showed that propane moved at a rate of 1223 μmol day⁻¹ from the leaves to the roots and 2652 μmol day⁻¹ from the roots to the leaves. These data demonstrated that gases diffuse rapidly through the plant body of T. latifolia.     

The Fate of 15N-Nitrate in Healthy and Declining Phragmites australis Stands

Abstract The dissimilatory nitrate-reducing processes, denitrification, and dissimilatory nitrate-reduction to ammonium were studied in freshwater lake sediments within healthy and degrading Phragmites australis (reed) stands. The samples from the healthy vegetation site contained roots and rhizomes. Cores were supplied with 1.9–5.2 μg ¹⁵N-NO₃ ⁻ g⁻¹ dry sediment in the laboratory and subsequently incubated for 8 h at 20°C, in the dark. The ¹⁵N compounds were determined before (natural percentage of ¹⁵N) and after 1 and 8 h of incubation. The uptake of ¹⁵N by the roots and rhizomes in the healthy vegetation was 61%. Nitrogen losses, interpreted as denitrification, accounted for 25 and 84% of the added ¹⁵N-NO₃ ⁻ in sediment from the healthy and degrading vegetation sites, respectively. The percentages of nitrate reduced to ammonium were 4 and 9% in sediment from the healthy vegetation and degrading vegetation sites, respectively. The percentage of ¹⁵N–total N in the sediment of the healthy vegetation site was 10%, whereas for the degrading vegetation site this percentage was 7%. The percentage of nitrate reduced to ammonium could be potentially underestimated by the percentage of ¹⁵N measured in the sediment. In this case, in healthy and degenerating P. australis stands, the percentage of produced ammonium accounted for 14–16%. The nitrate reduction rates were calculated based on an incubation period of one hour. The denitrification rate in sediment from the degrading vegetation site was higher than from the healthy vegetation site. The rate of dissimilatory nitrate reduction to ammonium was almost tenfold higher in sediment from the degrading vegetation site compared to sediment from the healthy vegetation site. The significantly lower percentages of dissimilatory nitrate reduction to ammonium and denitrification in the healthy stand compared to the degrading stand was probably due to the presence of roots and rhizomes. In the sediments of healthy and degrading P. australis stands, denitrification was the main nitrate-reducing process. Received: 24 July 1996; Accepted: 5 December 1996     

Effects of fire,mowing and nitrogen addition on root characteristics in tall-grass prairie

Abstract. Root harvests and root windows were used to study the influence of fire, mowing and nitrogen additions on root lengths, biomass, and nitrogen content in tall-grass prairie. Four years of nitrogen additions (10 g m² yr^?1) increased below-ground mass by 15 % and nitrogen concentration in that mass by 77 %. In general, live roots and rhizomes exhibited greater increases in nitrogen concentrations than detrital roots and rhizomes. After four years of treatment, live roots and rhizomes immobilized an additional 1.5 to 5 g/m² of nitrogen, depending upon specific treatment, while dead roots and rhizomes immobilized an additional 3 to 3.5 g/m². Average root growth parameters, as measured with root windows, were positively correlated with above-ground peak foliage biomass; however, the only significant correlation was between average new root growth and above-ground peak foliage biomass (r = 0.73, p ≤ 0.04). Root growth and decay, as measured by annual mean values for eight root windows over a four year interval, were insensitive to climatic and treatment effects.     

Seasonal variation in rates of heterotrophic nitrogen fixation (acetylene reduction) in Zostera noltii meadows and uncolonised sediments of the Bassin d'Arcachon, south-west France

Nitrogen fixation (acetylene reduction) rates were measured over an annual cycle in meadows of the seagrass Z. noltii and uncolonised sediments of the Bassin d'Arcachon, south-west France, using both slurry and whole core techniques. Measured rates using the slurry technique in Z. noltii colonised sediments were consistently higher than those determined in isolated cores. This was probably due to the release of labile organic carbon sources during preparation of the slurries. Thus, in colonised sediments the whole core technique may provide a more accurate estimate of in situ activity. Acetylene reduction rates measured by the whole core technique in colonised sediments were 1.8 to 4-fold greater, dependent upon the season, in the light compared with those measured in the dark, indicating that organic carbon released by the plant roots during photosynthesis was an important factor regulating nitrogen fixation. In contrast acetylene reduction rates in uncolonised sediments were independent of light.Addition of sodium molybdate, a specific inhibitor of sulphate reduction inhibited acetylene reduction activity in Z. noltii colonised sediments by > 80% as measured by both slurry and whole core techniques irrespective of the light regime, throughout the year inferring that sulphate reducing bacteria (SRB) were the dominant component of the nitrogen fixing microflora. A mutualistic relationship between Z. noltii and nitrogen fixing SRB in the rhizosphere, based on the exchange of organic carbon and fixed nitrogen is proposed. In uncolonised sediments sodium molybdate initially severely inhibited acetylene reduction rates, but the level of this inhibition declined over the course of the year. These data indicate that the nitrogen fixing SRB associated with the Zostera roots and rhizomes were progressively replaced by an aerobic population of nitrogen fixers associated with the decomposition of this recalcitrant high C:N ratio organic matter.Acetylene and sulphate reduction rates in the seagrass beds showed distinct summer maxima which correlated with a reduced availability of NH ₄ ⁺ in the sediment and the growth cycle of Z. noltii in the Bassin. Overall, these data indicate that acetylene reduction (nitrogen fixation) activity in the rhizosphere of Z. noltii was regulated both by release of organic carbon from the plant roots and maintenance of low ammonium concentrations in the root zone due to efficient ammonium assimilation.Nitrogen fixation rates determined from acetylene reduction rates measured by the whole core technique ranged from 0.1 to 7.3 mg N m^–2 d^–1 in the Z. noltii beds and between 0.02 and 3.7 mg N m^–2 d^–1 in uncolonised sediments, dependent upon the season. Nitrogen fixation in the rhizosphere of Z. noltii was calculated to contribute between 0.4 and 1.1 g N m^–2 y^–1 or between 6.3 and 12% of the annual fixed nitrogen requirement of the plants. Heterotrophic nitrogen fixation therefore represents a substantial local input of fixed nitrogen to the sediments of this shallow coastal lagoon and contributes to the overall productivity of Z. noltii in this ecosystem.     

Sulphate reduction and nitrogen fixation rates associated with roots, rhizomes and sediments from Zostera noltii and Spartina maritima meadows

Sulphate reduction rates (SRR) and nitrogen fixation rates (NFR) associated with isolated roots, rhizomes and sediment from the rhizosphere of the marine macrophytes Zostera noltii and Spartina maritima , and the presence and distribution of Bacteria on the roots and rhizomes, were investigated. Between 1‰ and 3‰ of the surface area of the roots and rhizomes of both macrophytes were colonized by Bacteria. Bacteria on the surfaces of S. maritima roots and rhizomes were evenly distributed, while the distribution of Bacteria on Z. noltii roots and rhizomes was patchy. Root- and rhizome-associated SRR and NFR were always higher than rates in the bulk sediment. In particular, nitrogen fixation associated with the roots and rhizomes was 41–650-fold higher than in the bulk sediment. Despite the fact that sulphate reduction was elevated on roots and rhizomes compared with bulk sediment, the contribution of plant-associated sulphate reduction to overall sulphate reduction was small (≤ 11%). In contrast, nitrogen fixation associated with the roots and rhizomes accounted for 31% and 91% of the nitrogen fixed in the rhizosphere of Z. noltii and S. maritima respectively. In addition, plant-associated nitrogen fixation could supply 37–1613% of the nitrogen needed by the sulphate-reducing community. Sucrose stimulated nitrogen fixation and sulphate reduction significantly in the root and rhizome compartments of both macrophytes, but not in the bulk sediment.     

Nitrogenase activity in the papyrus swamps of Lake Naivasha,Kenya

Nitrogenase activity (acetylene reduction activity) was found to occur universally in the Cyperus papyrus swamp in Lake Naivasha. Low rates of acetylene reduction activity (0.9–104.9 nmol C₂H₄ g d.wt. roots^-1 h^-1) were associated with excised roots of C. papyrus but higher rates of activity (89.0–280.4 nmol C₂H₄ g d.wt. roots^-1 h^-1) were associated with intact root systems of the plant. It was estimated that nitrogen fixation associated with young roots alone could supply about 26% of the nitrogen requirements of growing papyrus plants. Acetylene reduction activity in the lake bottom sediments was generally low and associated with adjacent papyrus stands. Plate counts of putative aerobic and facultatively anaerobic N₂-fixing bacteria associated with papyrus roots showed the presence of high numbers of diazotrophs (5.4 × 10⁶ CFU g d.wt. roots^-1). Fewer numbers of N₂-fixing bacteria were detected in the sediments (1.9 × 10³-3.2 × 10⁴ CFU g d.wt. sediment^-1).     

Comparison of the diversity of root-associated bacteria in Phragmites australis and Typha angustifolia L. in artificial wetlands

Common reed (Phragmites australis) and narrow-leaved cattail (Typha angustifolia L.) are two plant species used widely in artificial wetlands constructed to treat wastewater. In this study, the community structure and diversity of root-associated bacteria of common reed and narrow-leaved cattail growing in the Beijing Cuihu Wetland, China, were investigated using 16S rDNA library and PCR–denaturing gradient gel electrophoresis methods. Root-associated bacterial diversity was higher in common reed than in narrow-leaved cattail. In both plant species, the dominant root-associated bacterial species were Alpha, Beta and Gamma Proteobacteria, including the genera Aeromonas, Hydrogenophaga, Ideonella, Uliginosibacterium and Vogesella. Acidobacteria, Actinobacteria, Nitrospirae and Spirochaetes were only found in the roots of common reed. Comparing the root-associated bacterial communities of reed and cattail in our system, many more species of bacteria related involved in the total nitrogen cycle were observed in reed versus cattail, while species involved in total phosphorus and organic matter removal were mainly found in cattail. Although we cannot determine their nutrient removal capacity separately, differences in the root-associated bacterial communities may be an important factor contributing to the differing water purification effects mediated by T. angustifolia and P. australis wetlands. Thus, further work describing the ecosystem functions of these bacterial species is needed, in order to fully understand how effective common reed- and narrow-leaved cattail-dominated wetlands are for phytoremediation.     

Biological Nitrogen Fixation is not a Major Contributor to the Nitrogen Demand of a Commercially Grown South African Sugarcane Cultivar

It has previously been reported that endophytic diazotrophic bacteria contribute significantly to the nitrogen budgets of some graminaceous species. In this study the contribution of biological nitrogen fixation to the N-budget of a South African sugarcane cultivar was evaluated using ¹⁵N natural abundance, acetylene reduction and ¹⁵N incorporation. Plants were also screened for the presence of endophytic diazotrophic bacteria using acetylene reduction and nifH-gene targeted PCR with the pure bacterial strains. ¹⁵N natural abundance studies on field-grown sugarcane indicated that the plants did not rely extensively on biological nitrogen fixation. Furthermore, no evidence was found for significant N₂-fixation or nitrogenase activity in field-grown or glasshouse-grown plants using ¹⁵N incorporation measurements and acetylene reduction assays. Seven endophytic bacterial strains were isolated from glasshouse-grown and field-grown plants and cultured on N-free medium. The diazotrophic character of these seven strains could not be confirmed using acetylene reduction and PCR screening for nifH. Thus, although biological nitrogen fixation may occur in South African sugarcane varieties, the contribution of this N-source in the tested cultivar was not significant.     

Decomposition processes in soil of a healthy and a declining Phragmites australis stand

Decomposition processes were investigated in the soil of a declining, more eutrophic and a healthy, less eutrophic freshwater reed (Phragmites australis (Cav.) Trin. ex Steudel) stand in the littoral zone of Rožmberk fishpond, Czech Republic. Soil and pore water were sampled five times from April to October 1998. Chemical properties, CO₂ production in oxic and anoxic conditions, CH₄ production, denitrifying enzyme activity (DEA) and bacterial biomass were measured under laboratory conditions in suspensions prepared from homogenised soil samples. The more eutrophic West stand was more anaerobic than the East stand, with lower redox potential, lower pH and with a higher amount of organic acids, mainly acetic and lactic acid. Mean seasonal concentrations of total nitrogen in pore water, nitrogen of amino acids and proteins, and reducing sugars were all higher in the soil at the more eutrophic stand. Higher nutrient status and more reduced conditions at the more eutrophic stand were accompanied by (i) a limitation of aerobic microbial activities (CO₂ production in oxic conditions: 0.35 versus 0.54 μmol CO₂ cm⁻³ h⁻¹); lower DEA (4.0 versus 20.2 nmol N₂O cm⁻³ h⁻¹) and a lower proportion of bacteria that were active in aerobic conditions; (ii) by a prevalence of anaerobic over aerobic microbial processes; (iii) by a higher rate of methanogenesis (15.0 versus 11.5 nmol CH₄ cm⁻³ h⁻¹) and (iv) by an overall lower rate of microbial processes as compared to less eutrophied stand. The shift from aerobic to anaerobic microbial metabolism, and a coinciding restriction of metabolic activities at the more eutrophic stand are indicative of an elevated oxygen stress in the soil, associated with accumulation of metabolites toxic to both the micro-organisms and the reed. Possible links between eutrophication, decomposition processes in the soil and reed decline are discussed.     

N2(C2H2)ASE ACTIVITY BY ALNUS INCANA SSP. RUGOSA (BETULACEAE) IN THE NORTHERN HARDWOOD FOREST

Field assays of N₂(C₂H₂)ase activity were performed with intact nodules from a pure alder site (alder) and a mixed alder-aspen site (aspen). Assays were performed between 12 June and 12 August 1980 and in May 1981. N₂(C₂H₂)ase rates are expressed as g N g nodule oven-dry wt⁻¹ hr⁻¹ (g N g⁻¹ hr⁻¹). Diurnal N₂(C₂H₂)ase activity showed an increase in both sites between 0600 and midday, then decreased to a low by 1800. Nighttime activity in the May 1981 assay was approximately 25% of the daytime peak. Mean (±SE) 1200 hr N₂(C₂H₂)ase activity (μg N g⁻¹ hr⁻¹) for all sizes in the alder stand rose from 24.56 ± 6.56 on 12 June to 73.96 ± 28.37 on 26 June and declined to 9.20 ± 2.56 by 12 August. In the aspen stand activity decreased from the 12 June rate of 21.81 ± 4.59 to 3.64 ± 1.87 on 24 July but then increased to 30.00 ± 7.39 by 12 August. Based on diurnal assays, the seasonal mean N influx (μg N g⁻¹ hr⁻¹) is statistically higher (P 0.05) in the alder stand with a value of 26.70 compared to 14.63 in the aspen stand. Small size class shrubs had significantly higher (P < 0.05) N₂(C₂H₂)ase activity (μg N g⁻¹ hr⁻¹) in diurnal assays than medium or large class shrubs. The estimated mean (±SE) N₂(C₂H₂)ase activity (mg N g⁻¹ season⁻¹) for all sizes was 44.4 ± 18.6 in the alder stand compared to 16.2 ± 5.2 in the aspen stand. Nodule excavations showed the g shrub⁻¹ in the alder stand to be 16.48 ± 10.29, 38.57 ± 12.34 and 29.11 ± 7.15 for small, medium and large size shrubs and 12.73 ± 3.23, 28.21 ± 4.36 and 56.45 ± 16.23 for respective sizes in the aspen stand. Seasonal N influx was 4.69 kg ha⁻¹ in the alder stand and 0.84 kg ha⁻¹ in the aspen stand, representing 17.9% of the alder stand. Nitrogen feedback inhibition from uric acid-N influx and allelochemic interference from aspen are discussed as explanations for the differences in N influx in the two stands.     

Response of carbon and nitrogen content in plants and soils to vegetation cover change in alpine Kobresia meadow of the source region of Lantsang, Yellow and Yangtze Rivers

We conducted this study in lightly and severely degraded Kobresia pygmaea meadow in Gande County, Qinghai Province of China. The purpose of this research was to compare carbon and nitrogen concentrations, content and dynamics of aboveground tissue, belowground roots and soil (0-40 cm) between lightly and severely degraded Kobresia meadow. The results showed that C and N concentrations and C:N ratio of the aboveground tissue were significantly higher in lightly degraded grassland than in severely degraded grassland. In addition, total carbon and nitrogen concentrations of the aboveground tissue were ranked in order of forbs > grasses > sedges in the same grassland type. Total carbon and nitrogen concentrations of belowground roots were significantly higher in severely degraded grassland than in lightly degraded grassland. Total carbon and nitrogen concentrations were higher in the aboveground tissue than in the belowground roots. Total soil organic carbon concentration in severely degraded grassland was significantly lower than that in lightly degraded grassland, and decreased with depth. C and N content per unit area was ranked in order of 0-40 cm soil depth > belowground roots > aboveground issue in the same grassland type. The total carbon content per unit area of aboveground tissue, roots and 0-40 cm soil depth declined by 7.60% after degradation from lightly (14669.2 g m⁻²) to severely degraded grassland (13554.3 g m⁻²), i.e., 0-40 cm soil depth declined by 4.10%, belowground roots declined by 59.97% and aboveground tissue declined by 15.39%. The nitrogen content per unit area of aboveground tissue, roots and 0-40 cm soil depth increased after degradation by 12.76% from lightly (3352.7 g m⁻²) to severely degraded grassland (3780.6 g m⁻²), i.e., 0-40 cm soil depth increased by 13.07%, belowground roots declined by 55.09% and aboveground tissue declined by 16.00%. As a result of grassland degradation, the total carbon lost by 11149 kg hm⁻², and the total nitrogen increased by 4278 kg hm⁻².     

Impact of soil environmental factors on rates of N2-fixation associated with roots of intact maize and sorghum plants

We have evaluated the effects of oxygen partial pressure (pO₂), combined nitrogen, and the availability of organic substrates on nitrogen fixation (acetylene reduction) by bacteria associated with the roots of intact maize and sorghum plants. We also investigated the possibility of enhancing associative nitrogen-fixation by inoculating the soil in which the plants were grown withAzospirillum. Acetylene reduction (AR) activity was greatest when roots of intact plants were exposed to pO₂ between 1.3 and 2.1 kPa. Field-grown and greenhouse-grown plants supported similar levels of activity. Respiration inhibitors (2,4-dinitrophenol and sodium azide) eliminated AR activity at 2 kPa O₂, whereas a fermentation inhibitor (sodium fluoride) only partially reduced the activity. Acetylene reduction activity was rapidly (1–3 h) inhibited by NH ₄ ⁺ , NO ₃ ^– , and NO ₂ ^– at concentrations of 4–20 mg Nl^–1. Rates of AR varied substantially among individual plants in each experiment and between experiments. Amendment with any of several organic substrates greatly increased AR activity when rates were low, suggesting that the lack of activity was caused by a shortage of available carbon in the rhizosphere. Inoculation withAzospirillum failed to increase rates of AR associated with maize plants. In several experiments the indigenous bacteria associated with uninoculated plants exhibited greater activity than the bacteria associated with inoculated plants.     

Partitioning of nitrogen from biological fixation and fertilizer in Phaseolus vulgaris

Nitrogen uptake, distribution and remobilization in the vegetative and reproductive parts of the plant were studied in bean (Phaseolus vulgaris L.) cultivars Negro Argel and Rio Tibagi inoculated with either Rhizobium strain C05 or 127 K-17. Greenhouse grown plants were supplied with 2.5 mg N (plant)⁻¹ day⁻¹ as KNO₃ or K¹⁵NO₃ and the relative contribution to total plant nitrogen of mineral and symbiotically fixed nitrogen was determined. Control plants included those entirely dependent on fixed nitrogen as well as uninoculated plants supplied with 10 mg N (plant)⁻¹ day⁻¹. No differences were observed between inoculated treatments in total nitrate reductase activity and in the amount of mineral nitrogen absorbed, but there were considerable differences in the contribution of fixed nitrogen. Nitrogen fixation supplied from 58 to 72% of the total nitrogen assimilated during the bean growth cycle and the symbiotic combinations fixed most of their nitrogen (66 to 78% of total nitrogen) after flowering. Maximum uptake of mineral nitrogen was in the 15-day-period between flowering and mid-podfill (47 to 58% of total mineral nitrogen). Nitrogen partitioning varied with Rhizobium strains, and inoculation with strain C05 increased the nitrogen harvest index of both cultivars. Applied mineral nitrogen had a variable effect and in cv. Negro Argel was more beneficial to vegetative growth, resulting in smaller nitrogen harvest indices. Seed yield was not increased by heavy nitrogen fertilization. In contrast, cv. Rio Tibagi always benefited from nitrogen applications. Among the various nitrogen sources supplying the grain, the most important one was the fixed nitrogen translocated directly from nodules or after a rapid transfer through leaves, representing from 60 to 64% of the total nitrogen incorporated into the seeds.     

DISTRIBUTION AND PHYSIOLOGICAL DETERMINANTS OF BLUE-GREEN ALGAL NITROGEN FIXATION ALONG A THERMOGRADIENT1

The capacity of thermal algal-bacterial mats to fix nitrogen (N₂) was examined in an alkaline thermal stream, Rabbit Creek, of Yellowstone National Park. Nitrogenase activity and nitrogen-fixation rates of mat cores placed in serum bottles and incubated in situ were estimated by the acetylene-reduction technique. Active nitrogenase was not detected at 60 or 65 C in either the blue-green algal or bacterial undermat components of the mats. Acetylene was reduced by all mats ≤55 C along the thermogradient; mean fixation estimates for the mats ranged from 7 to 5,028 nmoles N₂ fixed · mg Chl a^?1· hr^?1. Maximum fixation occurred at 35 C in the stream; statistical comparison of mean rates ordered the thermogradient mats according to estimated activities: 35 > 40 > 30 > 50 ≥ 55 ≥ 45 C. Mats (≤40 C) dominated by species of Calothrix accounted for ca. 97% of the total nitrogen fixation observed in the stream; the remaining activity was associated with mats containing Mastigocladus laminosus Cohn. Light intensity significantly affected fixation rates of the Calothrix mats which responded in a linear fashion from 9–100% full sunlight (ca. 1,900 μEin · m^?2· sec^?1). Calothrix mats from 30 and 40 C had maximum nitrogenase activity at their growth temperature suggesting that nitrogen fixation along the thermogradient was optimally adapted to in situ temperatures.     

Root biomass,root distribution and the fine-root growth dynamics of Quercus coccifera L. in the garrigue of southern France

Quercus coccifera L., the characteristic scrub oak of the garrigue, covers more than 100,000 ha in southern France alone. Precipitation in this area averages 900 mm/year and summer rains are not rare. A total belowground biomass of 7.2 kg/m², including rhizomes and lignotubers, was harvested. Roots were concentrated in the uppermost 50 cm of the soil. It was hypothesized that low winter temperatures inhibit active fine-root growth. This hypothesis was tested by means of fine-root extractions of soil samples from 0–50 cm depth from November 1987 to June 1988. Although the fine-root analysis could not be extended into late summer and fall, the data supported the hypothesis. Ratios of live/dead fine roots reached their minimum at 0.2–0.3 from December to April. They increased to 1.0–1.2 during late spring and early summer. Initiation of fine-root growth in early April was synchronous with bud break. Starch contents of roots, rhizomes, and lignotubers fluctuated from 4.3% in January to 8.3% in April. The starch stored in belowground organs of Q. coccifera in a closed canopy stand amounted to about 500 g/m² in April. This amount declined to 400 g with bud burst and fine-root growth initiation.     

Expression of bacterial isochorismate synthase (EC 5.4.99.6) in transgenic root cultures ofRubia peregrina

Summary The gene coding for isochorismate synthase (EC 5.4.99.6) was amplified by the polymerase chain reaction fromEscherichia coli, cloned into a binary vector. and mobilised intoAgrobacterium rhizogenes LBA9402 which was used to transformRubia peregrina. Transgenic roots containing bacterial isochorismate synthase cDNA expressed twice as much isochorismate synthase activity (4.88 pkat/mg protein) as the control roots (2.45 pkat/mg protein) after 10 days in culture, and accumulatedca. 20% higher levels of total anthraquinones after 30 days in culture. Whilst the amount of total alizarin (free and bound) in the transgenic roots was 30% higher than in control roots, the level of free alizarin wasca. 85% higher.     

Comparison of nitrogen uptake in the roots and rhizomes of <Emphasis Type="Italic">Leymus chinensis</Emphasis>

Leymus chinensis (Trin.) Tzvel is a rhizomatous grass species in the Eastern Eurasian steppe zone that is often limited by low soil nitrogen availability. Although a previous study showed that the rhizomes of L. chinensis have the capacity to take up nitrogen, the importance of such uptake for nitrogen nutrition is unclear. Moreover, little is known regarding the inorganic nitrogen uptake kinetics of roots and rhizomes in response to nitrogen status. Here, we first found that ammonium is preferred over nitrate and glycine for L. chinensis growth. Using the ¹⁵N-labelling method, we found that the rate of ion influx into roots was approximately five-fold higher than into rhizomes under the same nitrogen content, and the ion influxes into roots and rhizomes under 0.05 mM N were greater than in the presence of 3 mM N, especially in the form of NH₄⁺. Using a non-invasive micro-test technique, we characterised the patterns of NH₄⁺ and NO₃^– fluxes in the root mature zone, root tip, rhizome mature zone, and rhizome tip following incubation in the solution with different N compounds and different N concentrations. These results suggest a dynamic balance between the uptake, utilisation, and excretion of nitrogen in L. chinensis.