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  • Articles  (234)
  • nitrogen fixation  (234)
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  • Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition  (234)
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  • Springer  (234)
  • De Gruyter
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Plant and soil 219 (2000), S. 285-289 
    ISSN: 1573-5036
    Keywords: actinorhizal plants ; Frankia ; 15N natural abundance ; nitrogen fixation ; Parasponia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Substantial enrichment of some plant parts in 15N relative to the rest of the plant is unusual, but is found in the nitrogen-fixing nodules of many legumes. A range of actinorhizal plants was surveyed to determine whether the nodules of any of them are also substantially enriched in 15N. The nonlegume Parasponia, nodulated by a rhizobium, was also included. Four of the actinorhizal genera and Parasponia were grown in N-free culture, and three actinorhizal genera were collected from the field. Nodules of Parasponia, Casuarina and Alnus were15N enriched relative to other plant parts, but only Parasponia approached the degree of enrichment found in some legume nodules. The nodules of Datisca, Myrica, Elaeagnus, Shepherdia, and Coriaria were depleted in 15N. Thus many actinorhizal nodules are depleted in 15N compared to other plant parts and enrichment is modest when it does occur. Whole plant 15N content (δ15N) in four actinorhizal plants and Parasponia showed a relatively narrow range of −1.41 to −1.90. Hence regardless of the degree of nodule enrichment or depletion, whole plant 15N content appears to vary little in plants grown in N-free culture.
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  • 2
    ISSN: 1573-5036
    Keywords: CO2 enrichment ; carbon storage ; climate change ; dissolved organic carbon ; nitrogen fixation ; root exudate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Root exudation has been hypothesized as one possible mechanism that may lead to increased inputs of organic C into the soil under elevated atmospheric CO2, which could lead to greater long-term soil C storage. In this study, we analyzed exudation of dissolved organic C from the roots of seedlings of the N-fixing tree Robinia pseudoacacia L. in a full factorial design with 2 CO2 (35.0 and 70.0 Pa) × 2 temperature (26° and 30 °C during the day) × 2 N fertilizer (0 and 10.0 mM N concentration) levels. We also analyzed the decomposition rates of root exudate to estimate gross rates of exudation. Elevated CO2 did not affect root exudation of organic C. A 4 °C increase in temperature and N fertilization did, however, significantly increase organic C exudation rates. Approximately 60% of the exudate decomposed relatively rapidly, with a turnover rate of less than one day, while the remaining 40% decomposed more slowly. These results suggest that warmer climates, as predicted for the next century, may accelerate root exudation of organic C, which will probably stimulate rapid C cycling and may make a minor contribution to intermediate to more long-term soil C storage. However, as these losses to root exudation did not exceed 1.2% of the net C fixed by Robinia pseudoacacia, root exudation of organic C appears to have little potential to contribute to long-term soil C sequestration.
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  • 3
    ISSN: 1573-5036
    Keywords: elevated CO2 ; FACE ; Lolium perenne L. ; N concentration ; nitrogen fixation ; plant litter ; species composition ; Trifolium repens L
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Experimental findings indicate that, in terrestrial ecosystems, nitrogen cycling changes under elevated partial pressure of atmospheric CO2 (pCO2). It was suggested that the concentration of N in plant litter as well as the amount of litter are responsible for these changes. However, for grassland ecosystems, there have been no relevant data available to support this hypothesis. Data from five years of the Swiss FACE experiment show that, under fertile soil conditions in a binary plant community consisting of Lolium perenne L. and Trifolium repens L., the concentration of litter N does not change under elevated atmospheric pCO2; this applies to harvest losses, stubble, stolons and roots as the sources of litter. This is in strong contrast to the CO2 response of L. perenne swards without associated legumes; in this case the above-ground concentration of biomass N decreased substantially. Increased symbiotic N2 fixation in T. repens nodules and a greater proportion of the N-rich T. repens in the community are regarded as the main mechanisms that buffer the increased C introduction into the ecosystem under elevated atmospheric pCO2. Our data also suggest that elevated atmospheric pCO2 results in greater amounts of litter, mainly due to increased root biomass production. This study indicates that, in a fertile grassland ecosystem with legumes, the concentration of N in plant litter is not affected by elevated atmospheric pCO2 and, thus, cannot explain CO2-induced changes in the cycling of N.
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  • 4
    Electronic Resource
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    Springer
    Agroforestry systems 47 (1999), S. 3-12 
    ISSN: 1572-9680
    Keywords: leguminous fallow ; nitrogen fixation ; soil fertility replenishment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Improved fallows are the deliberate planting of fast-growing species — usually legumes — for rapid replenishment of soil fertility. Improved fallows are rapidly spreading in several regions of the tropics as a sensible way for in situ accumulation of large quantities of N in vegetation and soil, as well as for providing sustainability enhancing services. Research on improved fallows increased after the mid 1980s with the development of what is known as the second soil fertility paradigm, which is based on sustainability considerations. Many lessons have emerged from short-term improved fallows (〈5 years duration). These include the diversity of farm sizes where improved fallows are used, the advantage of sequential versus simultaneous systems, the utilization of dry seasons unfavorable for crop production, the comparative advantages of woody versus herbaceous leguminous fallows, the magnitude of N accumulation, the strategic use of N fertilizers, and the importance of P. Other key services provided by fallows include fuelwood production, recycling of nutrients besides N, provision of a C supply to soil microorganisms, weed suppression, Striga control, and improved soil water storage. Natural fallows of non-legume shrubs belonging to the Asteraceae family, collectively called ‘daisy fallows’, may provide lessons for the development of improved fallows. The maintenance of genetic diversity in fallows is paramount. The main limiting factor in Africa is clearly the supply of germplasm of improved fallow species. This must be overcome though large-scale seed orchards and nursery development before impact at the scale of millions of farmers can take place.
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  • 5
    ISSN: 1573-5036
    Keywords: Galega orientalis ; nitrogen fixation ; polyamines ; Rhizobium galegae ; ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Plants of goat's rue (Galega orientalis) inoculated with Rhizobium galegae strain HAMBI 540 were grown in the presence of putrescine (Put), spermidine (Spd) or spermine (Spm), and several symbiotic characteristics were investigated to delineate the influence of polyamines (PA) on this nitrogen-fixing system. All three PA exerted a concentration-dependent effect on the nodule parameters tested. The increment of nodulation ability and nodule biomass accumulation was extreme (from 2.4- to 4.0-fold) when plants were subjected to 10 and 50 μM of various PA. However, at 100 μM a negative effect was observed. The acetylene-reduction activity of nodulated roots was increased also in response to treatment with the lower PA concentrations. The level of nitrogenase activity supported by succinate was significantly higher in bacteroids isolated from PA-treated nodules than in bacteroids from control nodules. The symbiotic parameters were also dependent on the type of PA used; the most effective being the diamine Put, while Spm showed a smaller physiological effect with respect to the others. Polyamines altered the ultrastructure of Galega nodule infected cells. After treatment with these substances, pronounced changes in the relative volume of the main components of infected cells and their compartments were observed. The significance of the structural observations and morphometric analyses, their relationship to differences in nitrogen fixation and possible modes of action are discussed.
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  • 6
    ISSN: 1573-5036
    Keywords: CO2 enrichment ; climate change ; dissolved organic nitrogen ; nitrogen cycle ; nitrogen fixation ; root exudate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract N-fixing trees facilitate the growth of neighboring trees of other species. These neighboring species benefit from the simple presence of the N fixation symbiosis in their surroundings. Because of this phenomenon, it has been hypothesized that a change in atmospheric CO2 concentration may alter the role of N-fixing trees in their environment. It is thought that the role of N-fixing trees in ecosystems of the future may be more important since they may help sustain growth increases due to increased CO2 concentration in nitrogen limited forests. We examined: (1) whether symbiotically fixed N was exuded from roots, (2) whether a doubled atmospheric CO2 concentration would result in increased organic N exudation from roots, and (3) whether increased temperature or N availability affected N exudation from roots. This study analyzed exudation of dissolved organic N from the roots of seedlings of the N-fixing tree Robinia pseudoacacia L. in a full factorial design with 2 CO2 (35.0 and 70.0 Pa) × 2 temperature (26 or 30 °C during the day) × 2 N fertilizer (0 and 10.0 mM N concentration) levels. Trees with no other source of N except N fixation exuded about 1% to 2% of the fixed N through their roots as dissolved organic N. Increased atmospheric CO2 concentrations did not, however, increase N exudation rates on a per gram belowground biomass basis. A 4 °C increase in temperature and N fertilization did, however, significantly increase N exudation rates. These results suggest that exudation of dissolved organic N from roots or nodules of N-fixing trees could be a significant, but minor, pathway of transferring N to neighboring plants in a much more rapid and direct way than cycling through death, decomposition and mineralization of plant residues. And, while exudation rates of dissolved organic N from roots were not significantly affected by atmospheric CO2 concentration, the previously observed ‘CO2 fertilization effect’ on N-fixing trees suggests that N exudation from roots could play a significant but minor role in sustaining increases in forest growth, and thus C storage, in a CO2 enriched atmosphere.
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  • 7
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    Nutrient cycling in agroecosystems 53 (1998), S. 71-81 
    ISSN: 1573-0867
    Keywords: fallow ; legumes ; nitrogen fixation ; rice ; weeds ; West Africa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Traditional upland rice-based cropping systems in West Africa rely on periods of fallow to restore soil fertility and prevent the build-up of insect pests and weeds. Demographic growth and increased demand for land is forcing many farmers to intensify their rice production systems. Declining fallow length and increasing number of crops before leaving the land to extended fallow result in a significant yield reduction. Promising cropping system alternatives include the use of site specific, weed-suppressing, multi-purpose cover legumes as short duration fallows. Constraints to rice production related to intensification were determined in 209 farmers' fields in four agro-ecological zones during 1994 and 1995. Nitrogen accumulation and weed suppression were evaluated in 54 legume accessions, grown for six months during the dry season, under a range of hydrological and soil conditions in 1994/95. Their effect on the yield of upland rice was determined in 1995. To increase benefits from improved fallow technology, the timing of legume establishment in relation to rice and the effect on crop and weed growth of removing, burning, mulching, or incorporating fallow residues prior to the rice crop were determined. Intensified land use resulted in a significant plot-level yield reduction that was highest in the derived savanna and the bimodal forest zones where it was associated with a doubling of the weed biomass in rice and a significant reduction in soil N supply. Legume fallows appear to offer the potential to sustain rice yields under intensified cropping. Legume biomass was in most instances significantly greater than in the weedy fallow control and several legume species suppressed weed growth. Nitrogen accumulation by legumes varied between 1–200 kg N ha-1 with 30–90% Ndfa. Rice grain yield following legume fallows increased by an average of 0.2 mg ha-1 or 29% above the weedy fallow control. Relay establishment substantially increased legume biomass. However, seeding of the legume at 28 days or earlier significantly reduced grain yield due to interspecific competition. Incorporating or mulching of fallow residues provided no significant yield advantage as compared to burning. Absolute effects varied as a function of site, legume species, and management practice.
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  • 8
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    Plant and soil 202 (1998), S. 41-47 
    ISSN: 1573-5036
    Keywords: Glycine max ; nitrogen fixation ; sulfur assimilation ; sulfur remobilization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract During pod filling, a grain legume remobilizes vegetative nitrogen and sulfur to its developing fruit. This study was conducted to determine whether different nitrogen sources affected N and S assimilation and remobilization during pod filling. Well-nodulated plants fed 1.0 mM KNO3, 0.5 mM urea, or 2.5 mM urea assimilated 0%, 37%, or 114% more N, respectively, and 25%, 46%, or 56% more S, respectively, than did the average non-nodulated control plant fed 5.0 mM KNO3. Thus, N source during pod filling greatly affected both N and S assimilation. Depending upon N source, plant N concentration during pod filling decreased from 2.96% to between 1.36% and 1.82%. Non-nodulated control plants fed 5.0 mM KNO3 had the highest residual N at harvest. During the same treatments, plant S concentration decreased from 0.246% to a relatively uniform 0.215%. Thus, during pod filling, vegetative N was seemingly remobilized more efficiently (38–54%) than was S (13%). N source also affected seed yield and seed quality. Non-nodulated control plants fed 5.0 mM KNO3 produced the lowest yield (21.1 g seeds plant-1), whereas well nodulated plants fed 1.0 mM KNO3, 0.5 mM urea, or 2.5 mM urea produced yields of 26.2 g, 31.8 g, and 36.7 g seeds plant-1, respectively. Non-nodulated plants fed 2.5 mM urea yielded 28.6 g of seeds plant-1. Seed N concentrations of non-nodulated plants and nodulated plants fed 2.5 mM urea were high, 6.30% and 6.11% N, respectively, whereas their seed S concentrations were low, 0.348% and 0.330% S, respectively. N sources that produced both a relatively high seed yield and seed N concentration (i.e., a relatively high total seed N plant-1) produced a proportionately smaller increase in total seed sulfur. Consequently, seed quality, as judged solely by seed S concentration, was lowered.
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  • 9
    ISSN: 1573-5036
    Keywords: Acetobacter diazotrophicus ; endophytic diazotrophic bacteria ; micropropagated plants ; nitrogen fixation ; sugar cane ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Micropropagated plantlets of sugar cane were inoculated with the N2-fixing bacterium Acetobacter diazotrophicus. Various modifications on the basic plant culture medium MS were made for the plant/bacteria association. The protocol required the inoculation of the bacteria at the end of the rooting period in a medium without hormones or vitamins, and with the concentration of sugar and mineral nutrients reduced by a factor of 10. Individual plants were inoculated with A. diazotrophicus and maintained under the appropriate light and temperature condition used for micropropagation up to 7 days. The system favored the infection and the establishment of the bacteria within the plant tissue. Bacteria colonized the plant tissue and accumulated in inter-cellular cavities and the region of lateral root emergence and also colonizes the xylem vessels. The inoculated plantlets were subsequently transferred to the acclimatization phase and after 30 days it was possible to isolate the bacteria from plant tissue. This protocol permitted studies of infection and comparison among strains.
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  • 10
    ISSN: 1573-5036
    Keywords: glutamine-synthetase ; leghaemoglobin ; nitrogen fixation ; nodulin ; PEP-carboxylase ; Phaseolus vulgaris ; Rhizobium tropici
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The metabolic activities of root nodules formed by Rhizobium tropici UM1899 were measured to test for the effects of geographical origin of the host bean (Phaseolus vulgaris L.) plant. Under increasing levels of N (0 to 24 mM of NH4NO3), the optimum condition for nitrogen fixation based on nitrogenase activity and allantoin concentration, was obtained between 2 and 4 mM N. Cultivars, including wild accessions from the two major domestication centers in America (Middle America and Andes), were then grown under aseptic conditions with 2 mM NH4NO3 and the rhizobial inoculant. Plant nodulins [leghaemoglobin (Lb), phosphoenolypyruvate carboxylase (PEPC) and glutamine synthetase (GS)], bacterial nitrogenase (NIF) activities as well as allantoin (ALA) concentration in the xylem sap, were assayed in flowering plants. Lb, PEPC, NIF activities and ALA concentrations were strongly affected by cultivar and by the center of origin. GS activity did not vary significantly with either cultivar or center of origin. LB, NIF and ALA were directly related to plant growth and offer opportunities to select for efficient N2-fixing symbioses. There were slight increases in nodulin activities of the domesticated cultivars, but the overall low variability within this material relative to landraces suggests that diversity for biological nitrogen fixation was reduced by domestication.
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  • 11
    ISSN: 1573-5036
    Keywords: celB ; gusA ; lacZ ; landrace ; lime ; molybdenum ; nitrogen fixation ; Phaseolus vulgaris ; Rhizobium tropici ; X-gal ; X-gluc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Poor bean yields in the Cunha region of the Mata Atlântica ecosystem in the state of São Paulo, Brazil, are associated with low agronomic inputs, plant disease, and soil erosion. To identify sustainable farming practices that increase production and maximize biological N2 fixation (BNF), the effects of soil fertility and plant cultivar on seed yield and root nodule formation were measured under standard agronomic practices. Results from 16 sites showed that fertilizing with lime and molybdenum increased seed yields to 370% for the landrace Serro Azul. In addition to increased yields, plants grown with fertilizer had more nodules. Marked strains of Rhizobium tropici were tested under controlled environments. An indicator strain of Rhizobium containing the gusA marker gene was used. Our results verify that the indicator strain CM-255 GusA+Hup+ had a high capacity to associate with the five bean varieties tested. Fertilization with P, K, S + micronutrients and liming were essential for better nodulation by the indicator strain. Under low fertility conditions, the landrace variety Serro Azul was poorly nodulated, when associated with native strains or with the indicator strain. However, under better soil fertility conditions, nodulation of Serro Azul by the marked Rhizobium strain was increased. The commercial variety Carioca 80SH showed no increase in nodulation (nodule number).
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  • 12
    ISSN: 1573-5060
    Keywords: inbred lines ; nitrate uptake ; nitrogen fixation ; nodules ; Trifolium repens ; white clover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract N2 fixation, nitrate uptake, and tissue N levels werecompared in four novel self-fertile inbred lines (referred to as A, B, C andD) of white clover (Trifolium repens L.) believed to differ markedly intheir nitrogen relations. Plants were inoculated with a mixture of Rhizobiumstrains and grown in flowing nutrient solutions without mineral N for 18 dprior to supplying half the plants with 20 µM $$NO_3^ - $$ over a 36 d treatment period. Net uptakeof $$NO_3^ - $$ was measured automatically on a dailybasis. The remaining plants constituted ‘controls’ solely dependent uponN2 fixation for acquisition of N. Total uptake of $$NO_3^ - $$ over 36 days was 27.4 (A), 23.8 (B), 13.8(C) and 30.1 (D) mmol N/plant, compared with N2 fixation of7.2 (A), 16.9 (B), 26.2 (C) and 6.2 (D) mmol N/plant by the $$NO_3^ - $$ -fed lines. Total N2 fixationby corresponding ‘control’ plants was 15.6 (A), 35.6 (B), 35.3 (C) and 2.2(D) mmol N/plant. Line D plants exhibited a 3:1 segregation with respect toeither ‘nil’ or ‘partially impaired’ N2 fixation; thecorresponding ratio for line A was 1:3. Line C plants had a ‘low $$NO_3^ - $$ uptake’ phenotype characterised by specific $$NO_3^ - $$ uptake rates approximately half those ofthe other lines, higher N2 fixation in the presence of $$NO_3^ - $$ , and low leaf + petiole concentrations of $$NO_3^ - $$ , although concentrations of $$NO_3^ - $$ in stolon and root fractions were similarin all lines. These inbred lines provide a tool for (1) the analysis of thegenetic and physiological basis of variation in the efficacy with which $$NO_3^ - $$ uptake down-regulates N2fixation, and (2) testing ecological hypotheses for temporal and spatialvariation in clover/grass coexistence in pastures.
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  • 13
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    Plant and soil 198 (1998), S. 63-69 
    ISSN: 1573-5036
    Keywords: allocation ; Alnus ; nitrogen ; nitrogen fixation ; resorption ; translocation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Red alder (Alnus rubra Bong.) trees were labeled with15 NO- 3 or15 NH+ 4 using the stem-injection method. Leaves were sampled 3 and 15 months subsequent to injection within several crown positions, including top, bottom, proximal, medial, and distal. Stem injection of both15 NH+ 4 and15 NO- 3 at levels approaching 1% of crown N effectively labeled red alder trees. Although more variable, 15NO- 3 may have been more efficient in initial labeling. The distribution of 15N in the crown was uniform 3 months after labeling, but was diluted in the distal and top positions by the following year. In both years there was a clear increase in total N concentration toward the periphery of the tree. This increase became more pronounced with increasing crown size and crown closure. Crown position with respect to light availability may be the most important determinant in N allocation in red alder foliage.
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  • 14
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    Plant and soil 189 (1997), S. 181-188 
    ISSN: 1573-5036
    Keywords: Bradyrhizobium ; legume ; nitrogen fixation ; soybean ; symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Several genes that restrict nodulation with specific Bradyrhizobiumstrains are known in Glycine max (soybean), and a similar system of nodulation restriction has recently been discovered in the related North American legume Amphicarpaea bracteata. We analyzed how nodulation-restrictive genotypes of each plant interacted with Bradyrhizobium strains sampled from the other host species. Ten bacterial isolates from A. bracteata that nodulated differentially with genotypes of their homologous host legume showed uniform responses to two soybean isogenic lines that differed at the Rj4 locus controlling nodulation restriction: all isolates formed nodules of normal size and morphology on both isolines. However, little or no nitrogen fixation occurred in any of these symbioses. A. bracteata genotypes that displayed broad vs. restricted symbiotic phenotypes toward naturally-associated bradyrhizobia were also tested with two bacterial isolates from soybean (USDA 76 and USDA 123). Both isolates formed nodules and fixed nitrogen in association with both A. bracteata genotypes. However, symbiotic effectiveness (as measured by acetylene reduction assays) was normal only for the combination of USDA 76 with the restrictive A. bracteata genotype. Overall, these results indicate that plant genes that restrict nodulation by certain naturally-associated bradyrhizobia do not confer comparable specificity when plants interact with bacteria from another related legume species.
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  • 15
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    Plant and soil 189 (1997), S. 221-229 
    ISSN: 1573-5036
    Keywords: azide resistance ; nitrogen fixation ; phenethyl alcohol ; t Rhizobium loti ; TMPD-oxidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Azide-resistant (AzR) mutants of Rhizobium loti strain NZP2037 were isolated. Mutations conferring azide resistance (azi) appeared at a frequency of 0.5 × 10-7. Nine AzR mutants of R. loti were characterised for their symbiotic behaviour with Lotus pedunculatus plants. In comparison to the wild type parent strain, AzR mutants exhibited either similar or higher symbiotic effectiveness. The azi mutations which enhanced nitrogen fixation as well as improving shoot dry weight of the inoculated plants also increased nodulation. Unlike several azi mutations in Escherichia coli, these azi mutations did not alter sensitivity of R. loti to phenethyl alcohol. One of the AzR mutants exhibited higher micro-aerobic, N′, N′, N′, N′-tetramethyl-p-phenylenediamine (TMPD) oxidase activity.
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  • 16
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    Plant and soil 189 (1997), S. 205-211 
    ISSN: 1573-5036
    Keywords: amelioration ; coastal salinity ; halotolerant cyanobacteria ; “Kharland” soils ; nitrogen fixation ; soil reclamation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A brackish-water, nitrogen-fixing cyanobacterium, Anabaena torulosa, could successfully grow and fix nitrogen on moderately saline “Kharland” soils (soil conductivity 5 to 8.50 dS m-1), typical of Indian coastline. During five weeks of growth under laboratory as well as field conditions, the cyanobacterium exhibited high rates of nitrogen fixation and substantially enriched the nitrogen status of saline soils (43-76%), although the fixed nitrogen remained confined to the cyanobacterial biomass. Most (〉90%) of the cell-bound Na+ remained extracellularly trapped in the mucopolysaccharide sheath of A. torulosa; traces of the cation that permeated cyanobacterial cells were found to exist in an osmotically active, free state. No evidence was found for the incorporation of Na+ into any biomolecule, especialty proteins or carbohydrates. Therefore, permanent removal of Na+ from saline soils using cyanobacteria may not be possible, since Na+ is released back into the soil subsequent to the death and decay of cyanobacteria. Removal of top soil containing cyanobacterial mats significantly decreased the soil salinity (between 26-38%). But such a practice removes all the fixed nitrogen and carbon and also does not seem feasible on a large scale. Amelioration of soil salinity by simultaneous application of A. torulosa during crop growth seems to be an attractive possibility, especially since it can also supplement the nitrogen requirement of the crop.
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  • 17
    ISSN: 1573-5036
    Keywords: ammonium ; Azotobacter ; diazotroph ; glutamine synthetase ; NifA ; nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Strategies considered and studied for achieving ammonium excretion in nitrogen fixing bacteria include 1) inhibition of ammonium assimilation and 2) interference with the mechanisms by which ammonium inhibits either nitrogenase synthesis or activity. These aspects of nitrogen fixation have been best studied in diazotrophic Proteobacteria and Cyanobacteria and those of the former are reviewed in this paper. Ammonium assimilation by glutamine synthetase (GS) can be diminished or prevented by treatment of bacteria with chemicals that inhibit GS activity and in some diazotrophs, such treatment results in excretion of up to 15mM ammonium into liquid growth medium. Also, mutants with altered GS activity, isolated by selection for resistance to GS inhibitors, often excrete ammonium. In Proteobacteria, ammonium inhibits nitrogenase activity and/or synthesis, the latter by preventing activity or expression of NifA, a transcriptional activator required for expression of other nif genes. In Azotobacter vinelandii, ammonium inhibits NifA activity but not its synthesis; NifL mediates this effect by interacting directly with NifA causing its inactivation. In nifL insertion mutants, NifA is constitutively active and up to 10 mM ammonium is excreted during nitrogen fixation. GlnD insertion/deletion mutations are unable to be stably maintained in A. vinelandii wild type but are stable and viable in a mutant that produces constitutively active GS (cannot be adenylylated). This confirms the hypothesis that GlnD is required for activity of GS, an essential enzyme in A. vinelandii. In addition, the stable glnD mutants are Nif, supporting also the previous conclusion that GlnD is involved in mediating NifL/NifA interaction. Mechanisms of inhibition of synthesis or activity of NifA by ammonium in other diazotrophs are discussed and compared.
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  • 18
    ISSN: 1573-5036
    Keywords: gibberellic acid ; Glycine max ; grain yield ; nitrogen fixation ; protein yield ; short season
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In short-season soybean production areas, low soil temperature is the major factor limiting soybean establishment, nodulation and nitrogen fixation. Gibberellic acid (GA) pretreatment of crop seeds can overcome low soil temperature inhibition of seed germination and seedling development. However, previous studies have found that the application of GAs decreased legume nodulation and nitrogen fixation under optimal growth conditions. A field experiment was conducted under short season conditions in eastern Canada to determine whether the application of GA3 to soybean seed could accelerate germination, and increase plant nodulation and nitrogen fixation. The results indicated that GA3 application accelerated seedling emergence but decreased plant nodulation and nitrogen accumulation at early plant growth stages. However, these initial negative effects were overcome as the plants developed. Gibberellic acid applied to soybean seed at the time of planting did not influence final grain and protein yield.
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  • 19
    ISSN: 1573-5036
    Keywords: acetylene reduction ; acetylene-induced decline ; argon-induced decline ; actinorhizal plants ; Elaeagnus ; Frankia ; nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The rate of C2H2 reduction by nodulated seedlings of Elaeagnus angustifolia (Russian olive) was followed as a function of time. Our goals were to: 1) determine whether there is an C2H2-induced decline in nitrogenase activity; and 2) investigate the mechanism of any decline. We found a peak rate of C2H2 reduction at 1.5 min after the introduction of C2H2 that was followed by a rapid decline in activity to 56% of the peak value. After the decline there was a partial recovery to 67% of the peak value at 60 min. When the pO2 was decreased during the decline there was no significant effect (p≤0.05) on nitrogenase activity. When the C2H2 reduction assay was preceded by an incubation in a gas mixture (20 kPa O2) with Ar substituted for N2, there was little decline in nitrogenase activity as a function of time, but the rate of C2H2 reduction per gram nodule was reduced by approximately 50%. From these results we conclude that t Elaeagnus angustifolia exhibits a pronounced C2H2-induced decline and consequently the initial peak rate C2H2 reduction must be determined to obtain a valid measure of nitrogenase activity. We further suggest that cessation of NH3 formation initiates the decline and that the decline is not caused by a change in nodule permeability to gases.
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  • 20
    ISSN: 1573-5036
    Keywords: Bradyrhizobium jopanicum ; genistein ; Glycine max ; low soil temperature ; nitrogen fixation ; nodulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In the soybean (Glycine max. (L.) Merr)– Bradyrhizobium japonicum symbiosis, suboptimal root zone temperatures (RZTs) slow nodule development by disruption of the interorganismal signal exchange between the host plant and bradyrhizobia. Two field experiments were conducted on two adjacent sites in 1994 to determine whether the incubation of B. japonicum with genistein prior to application as an inoculant, or genistein, without B. japonicum, applied onto seeds in the furrow at the time of planting, increased soybean nodulation, N fixation, and total N yield. The results of these experiments indicated that genistein application increased nodule number and nodule dry matter per plant and hastened the onset of N fixation during the early portion of the soybean growing season, when the soils were still cool. Because these variables were improved, total fixed. N, fixed N as a percentage of total plant N, and N yield increased due to genistein application. The interaction between genistein application and soybean cultivars indicated that genistein application was more effective on N-stressed plants.
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  • 21
    ISSN: 1573-5036
    Keywords: actinorhizal plants ; aluminium toxicity ; Casuarina cunninghamiana ; Frankia ; nitrogen fixation ; nodulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In order to investigate the effects of Al on nodule formation and function in the Casuarina-Frankia symbiosis, inoculated plants were grown in sand culture at five nominal Al concentrations (0-880 μM Al) at pH 4.0. There was an Al-free control at pH 6.0 to assess the effects of pH 4.0 treatments. Mean N concentration of nodules was significantly less at pH 4.0 (1.83%) than at pH 6.0 (2.01%). There were nodulated plants at all Al levels, though there were fewer nodulated plants at 440 and 880 μM Al. Dry weights of nodules, shoots and roots were not reduced by Al concentrations at or below 220 μM Al, but were decreased by Al concentrations at or above 440 μM Al. Nodule weight expressed as a percentage of total weight did not differ significantly with respect to an Al-free control at pH 4. N concentrations of shoots and whole plants were significantly reduced at 440 μM Al. Nodular specific acetylene reduction activity (ARA) did not differ significantly among Al treatments. However, N2-fixation efficiency was decreased from 0.20 to 0.10 mg N fixed mg nodule dry weight−1 at 880 μM Al.
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  • 22
    ISSN: 1573-5036
    Keywords: endophytic microbes ; endo-symbiosis ; genomic fingerprinting ; infection ; marker genes ; nitrogen fixation ; PCR ; rep-PCR ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The extension of nitrogen-fixing symbioses to important crop plants such as the cereals has been a long-standing goal in the field of biological nitrogen fixation. One of the approaches that has been used to try to achieve this goal involves the isolation and characterization of stable endophytic bacteria from a variety of wild and cultivated rice species that either have a natural ability to fix nitrogen or can be engineered to do so. Here we present the results of our first screening effort for rice endophytes and their characterization using acetylene reduction assays (ARA), genomic fingerprinting with primers corresponding to naturally occurring repetitive DNA elements (rep-PCR), partial 16S rDNA sequence analysis and PCR mediated detection of nitrogen fixation (nif) genes with universal nif primers developed in our laboratory. We also describe our efforts to inoculate rice plants with the isolates obtained from the screening, in order to examine their invasiveness and persistence (stable endophytic maintenance). Lastly, we review our attempts to tag selected isolates with reporter genes/proteins, such as beta-glucuronidase (gus) or green fluorescent protein (gfp), in order to be able to track putative endophytes during colonization of rice tissues.
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  • 23
    ISSN: 1573-5036
    Keywords: Acetobacter ; auxin ; 2 ; 4-D ; clover ; nitrogen fixation ; Rhizobium ; rice ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Root morphology is both genetically programmed and environmentally determined. We have begun an analysis into the components of root development by: (a) constructing a range of transgenic clover plants to assess some of the genetic programs involved as both roots and nodules are initiated and develop. These transgenic plants report on auxin activity, flavonoid synthesis and chitinase expression and suggest a role for flavonoids as regulators of auxin levels; and (b) determining in cereals the effect of both added auxin and specific microorganisms on the initiation and development of modified root outgrowths and lateral roots. Appropriate combinations of auxin, the nitrogen fixing Acetobacter diazotrophicus, and rice variety did give rise to some plants which grew slowly for over 12 months in a nitrogen-free medium.
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  • 24
    ISSN: 1573-5036
    Keywords: actinorhizal plants ; evolution ; nitrogen fixation ; phylogenetic hypotheses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Current taxonomic schemes place plants that can participate in root nodule symbioses among disparate groups of angiosperms. According to the classification scheme of Cronquist (1981) which is based primarily on the analysis of morphological characters, host plants of rhizobial symbionts are placed in subclasses Rosidae and Hamamelidae, and those of Frankia are distributed among subclasses Rosidae, Hamamelidae, Magnoliidae and Dilleniidae. This broad phylogenetic distribution of nodulated plants has engendered the notion that nitrogen fixing endosymbionts, particularly those of actinorhizal plants, can interact with a very broad range of unrelated host plant genotypes. New angiosperm phylogenies based on DNA sequence comparisons reveal a markedly different relationship among nodulated plants and indicate that they form a more coherent group than has previously been thought (Chase et al., 1993; Swensen et al., 1994; Soltis et al., 1995). Molecular data support a single origin of the predisposition for root nodule symbiosis (Soltis et al., 1995) and at the same time support the occurrence of multiple origins of symbiosis within this group (Doyle, 1994; Swensen, 1996; Swensen and Mullin, In Press).
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  • 25
    ISSN: 1573-5036
    Keywords: Azospirillum brasilense ; evolution ; nifA-lacZ ; nifH-lacZ ; nitrogen fixation ; para-nodules ; symbiosis ; wheat ; 2 ; 4-D
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Recent advances towards achieving significant nitrogen fixation by diazotrophs in symbioses with cereals are reviewed, referring to the literature on the evolution of effective symbioses involving rhizobia and Frankia as microsymbionts. Data indicating that strains of Acetobacter and Herbaspirillum colonizing specific cultivars of sugarcane as endophytes make a significant contribution to the nitrogen economy of this crop improves the prospects that similar associative systems may be developed for other gramineous species such as rice and wheat. By contrast, the transfer of nodulation genes similar to those in legumes or Parasponia to achieve nodulation in crops like rice and wheat is considered to be a more ambitious and distant goal. Progress in developing an effective associative system for cereals has been materially assisted by the development of genetic tools based on the application of lacZ and gusA fusions with the promoters of genes associated with nitrogen fixation. These reporter genes have provided clear evidence that ‘crack-entry’ at the points of emergence of lateral roots or of 2,4-D induced para-nodules is the most significant route of endophytic colonization. Furthermore, using the laboratory model of para-nodulated wheat, there is now evidence that the ability of azospirilla and other nitrogen fixing bacteria to colonize extensively as endophytes can be genetically controlled. The most successful strain of Azospirillum brasilense (Sp7-S) for endophytic colonization and nitrogen fixation in wheat seedlings is a mutant with reduced exopolysaccharide production. Most other strains of azospirilla do not colonize as endophytes and it is concluded that though these are poorly adapted to providing nitrogen for the host plant, they are well adapted for survival and persistence in soil. A research program combining the study of endophytic colonization by azospirilla with an examination of the factors controlling the effectiveness of association (oxygen tolerance and nitrogen transfer) is now being pursued. It is proposed that a process of facilitated evolution of para-nodulated wheat involving the stepwise genetic improvement of both the prospective microsymbionts and the cereal host will eventually lead to effective nitrogen-fixing associations. In the attempt to achieve this goal, continued study of the endophytes occurring naturally in sugar cane and other grasses (e.g. Azoarcus sp.) should be of assistance.
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  • 26
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    Plant and soil 194 (1997), S. 1-10 
    ISSN: 1573-5036
    Keywords: endophyte ; legume-Rhizobium ; nif gene ; nitrogen fixation ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Recent advances in understanding symbiotic Rhizobium-legume interactions at the molecular level, the discovery of endophytic interactions of nitrogen-fixing organisms with non-legumes, and the ability to introduce genes into rice by transformation have stimulated researchers world wide to harness opportunities for nitrogen fixation and improved N nutrition in rice. In a think-tank workshop organized by IRRI in 1992, the participants reaffirmed that such opportunities do exist for cereals and recommended that rice be used as a model system. Subsequently, IRRI developed a New Frontier Project to coordinate the worldwide collaborative efforts among research centers committed to reducing dependency of rice on mineral N resources. An international Biological Nitrogen Fixation (BNF) working group was established to review, share research results/materials and to catalyze research. The strategies of enabling rice to fix its own N are complex and of a long-term nature. However, if achieved, they could enhance rice productivity, resource conservation, and environmental security. The rate of obtaining success would, of course, benefit tremendously from concerted efforts from a critical mass of committed scientists around the world, as well as a constant and continued funding support from the ‘donor’ community.
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  • 27
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    Plant and soil 194 (1997), S. 57-64 
    ISSN: 1573-5036
    Keywords: Azoarcus ; endorhizosphere ; nif-genes ; nitrogen fixation ; 16S ribosomal RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The current knowledge on the divergence within the genus Azoarcus and about interactions with grasses is summarized. Grass-associated members of this genus of diazotrophs have only been isolated from a salt- and flood-tolerant pioneer plant in Pakistan, Kallar grass (Leptochloa fusca (L.) Kunth). Members of these bacteria belong to the beta subclass of the Proteobacteria, most closely related to purple bacteria such as Rhodocyclus purpureus. The isolates from one single host plant showed a surprising divergence, consisting of five groups of Azoarcus distinct at the species level. Molecular diagnostic tests, which are based on 16S ribosomal RNA sequences, allowed preliminary assignment of isolates to Azoarcus by PCR amplification and sequencing of PCR products. Moreover, the moleculer tests enabled us to detect an unculturable strain in Kallar grass roots, stressing that classical cultivation techniques at times fail to detect some groups of the microbial population. Using similar techniques, sequences rooting in the Azoarcus clade were also detected in field-grown rice, indicating that the natural host range might extend to rice. In gnotobiotic laboratory cultures, a member of Azoarcus is able to colonize rice roots endophytically: bacteria invade the roots in the zone of elongation and differentiation, colonize the cortex intra- and inter-cellularly, and penetrate deeply into the vascular system, entering xylem vessels, allowing systemic spreading into the rice shoot. Recently, we detected expression of nitrogenase of Azoarcus cells inside roots of rice seedlings, a result encouraging us to analyze interactions with rice in detail.
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  • 28
    ISSN: 1573-5036
    Keywords: competition ; drought ; Medicago falcata ; Medicago sativa ; nitrogen fixation ; nodulation ; rhizobia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Drought is an important environmental factor that can affect rhizobial competition and N2 fixation. Three alfalfa (Medicago sativa L. and M. falcata L.) accessions were grown in pots containing soil from an irrigated (Soil 1) and a dryland (Soil 2) alfalfa field in northern Utah, USA. Mutants of three strains of Rhizobium meliloti Dang. from Pakistan (UL 136, UL 210, and UL 222) and a commercial rhizobial strain 102F51a were developed with various levels of resistance to streptomycin. Seeds inoculated with these individual streptomycin-resistant mutants were sown in the two soils containing naturalized rhizobial populations. Soils in the pots were maintained at −0.03, −0.5, and −1.0 MPa. After 10 weeks, plants were harvested and nodule isolates were cultured on agar medium with and without streptomycin to determine nodule occupancy (proportion of the nodules occupied by introduced rhizobial strains). Number of nodules, nodule occupancy, total plant dry weight, and shoot N were higher for Soil 1 than Soil 2. Number of nodules, plant dry weight, and shoot N decreased as drought increased from −0.03 to −1.0 MPa in the three alfalfa accessions. Rhizobial strains UL 136 and UL 222 were competitive with naturalized alfalfa rhizobia and were effective at symbiotic N2 fixation under drought. These results suggest that nodulation, growth, and N2 fixation in alfalfa can be improved by inoculation with competitive and drought-tolerant rhizobia and may be one economically feasible way to increase alfalfa production in water-limited environments.
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  • 29
    ISSN: 1573-5036
    Keywords: cultivar, grazing ; 15N ; nitrogen fertilizer ; nitrogen fixation ; Trifolium repens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Forage production and N2 fixation were determined for nine cultivars of white clover (Trifolium repens L.) grown with perennial ryegrass (Lolium perenne L.) and receiving nitrogen (N) fertilizer at either 0 or 390 kg N ha-1 yr-1. The site was grazed by sheep at 3 or 6 (in winter and summer) week intervals and N fertilizer was applied at 30 kg N ha-1 after each grazing. Annual white clover production showed a 2-fold variation between cultivars and was positively correlated with total pasture production in the 0 N treatment. Nitrogen fertilizer application increased average total pasture dry matter (DM) production from 12830 to 16010 kg ha-1 yr-1, but decreased average white clover production from 3600 to 2970 kg DM ha-1 yr-1. Fertilizer N application decreased annual N2 fixation from 111 to 47 kg N ha-1 (mean for all cultivars, using 15N dilution), with the decline occurring predominantly in spring and summer. The decrease in N2 fixation in spring from 47 to 18 kg N ha-1 was due largely to a decrease in clover DM production. In contrast, N application decreased average N2 fixation in summer from 36 to 14 kg N ha-1 due mainly to a large decrease in the proportion of clover N derived from atmospheric N2 (from 49 to 24%), with clover DM production falling by only 10%. Clover cultivars showed a variation in annual N2 fixation of about 3-fold under both N regimes. During winter and spring, the amount of N fixed by the different cultivars was determined predominantly by their DM production in both N regimes. In contrast, during summer and autumn there was a marked variation between cultivars in tolerance of N2 fixation to increased soil inorganic N due to N fertilizer application. This was evident from a significant cultivar × N interaction for the proportion (PN) of clover N derived from N2 fixation. During summer/autumn, PN for Kopu was similar in the 0 and 390 N treatments, whereas PN declined by up to two-thirds for the other cultivars. Consequently, in summer/autumn the amount of N fixed by Kopu decreased by only 20% (from 65 to 52 kg N ha-1) due to N application whereas it decreased by 40–80% (to 15–34 kg N ha-1) for the other cultivars. Thus, N2 fixation during winter/spring was highest for the most productive large-leaved cultivars (Kopu, Aran and Pitau) either in the absence or presence of added N. In contrast, in summer/autumn the cultivars differed in tolerance to added N, and N2 fixation in the 390 N treatment was higher for the more tolerant cultivar Kopu than for the other cultivars.
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  • 30
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    Plant and soil 178 (1996), S. 215-222 
    ISSN: 1573-5036
    Keywords: drought stress ; gnotobiotic assembly ; Mimoseae ; nitrogen fixation ; organic sulfide ; plant age ; sulfate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Carbon disulfide (CS2) and carbonyl sulfide (COS) are colorless, foul-smelling, volatile sulfur compounds with biocidal properties. Some plants produce CS2 or COS or both. When used as an intercrop or forecrop, these plants may have agronomic potential in protecting other plants. Most of the factors which affect production of these plant-generated organic sulfides are unknown. We determined the effects of sulfate concentration, plant age, nitrogen fixation, drought stress, root injury (through cutting), and undisturbed growth on COS production in Leucaena retusa or Leucaena leucocephala and the effect of some of these factors on CS2 production in Mimosa pudica. In addition, we determined if organic sulfides were produced in all Leucaena species. When L. retusa and M. pudica seedlings were grown in a plant nutrient medium with different sulfate concentrations (50 to 450 mg SL-1), COS or CS2 from crushed roots generally increased with increasing sulfate concentration. COS production was highest (≤74 ng mg-1 dry root) for young L. retusa seedlings and declined to low amounts (〈5 ng mg-1 dry root) for older seedlings. Nitrogen fixation reduced the amounts of COS or CS2 produced in L. leucocephala and M. pudica. Under conditions of undisturbed growth, root cutting, or drought stress, no COS production was detected in 4-to 8-weeks-old L. retusa plants. COS or CS2 or both was obtained from crushed roots or shoots of all 13 known Leucaena species.
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  • 31
    ISSN: 1573-5036
    Keywords: competitiveness ; inoculant ; nitrogen fixation ; nodulation ; soybean ; yield
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Bradyrhizobium japonicum strain TA-11NOD+, with altered indole biosynthesis, exhibited enhanced nodulation and nitrogen fixation on soybean in previous greenhouse studies. In this study, field experiments were conducted at Upper Marlboro, Maryland, in the summers of 1988 and 1993. In 1988, the site used was essentially free of soybean-nodulating bacteria and seed yield in plots inoculated with either I-110ARS or TA-11NOD+ was significantly higher by 12 or 20%, respectively, than that of the uninoculated controls. The 1993 site had an indigenous soil population (about 104 cells g-1) of symbiotically ineffective soybean-nodulating bacteria. Nevertheless, six-week-old ‘Morgan’ soybean plants inoculated with strain TA-11NOD+ had 44% more nodules and exhibited 50% more nitrogen fixation by acetylene reduction when compared with plants that received the parental strain I-110ARS. Nodule occupancy, as determined using genetic markers for rifampicin and streptomycin resistance, was significantly higher for strain TA-11NOD+ than for strain I-110ARS. Overall, for the two years and the two soybean genotypes, the yield obtained with TA-11NOD+ was 6% higher than that obtained with I-110ARS. Competition experiments were conducted in the greenhouse and strain TA-11NOD+ was significantly more competitive than strain I-110ARS in competition with strains USDA 6 or USDA 438.
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  • 32
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    Plant and soil 186 (1996), S. 173-187 
    ISSN: 1573-5036
    Keywords: Bradyrhizobium ; legumes ; nitrogen fixation ; Nod Factors ; nodulation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Genes controlling nitrogen-fixing symbioses of legumes with specialized bacteria known as rhizobia are presumably the products of many millions of years of evolution. Different adaptative solutions evolved in response to the challenge of survival in highly divergent complexes of symbionts. Whereas efficiency of nitrogen fixation appears to be controlled by quantitative inheritance, genes controlling nodulation are qualitatively inherited. Genes controlling nodulation include those for non-nodulation, those that restrict certain microsymbionts, and those conditioning hypernodulation, or supernodulation. Some genes are naturally occurring polymorphisms, while others were induced or were the result of spontaneous mutations. The geographic patterns of particular alleles indicate the role of coevolution in determining symbiont specificites and compatibilities. For example, the Rj4 allele occurs with higher frequency (over 50%) among the soybean (G. max) from Southeast Asia. DNA homology studies of strains of Bradyrhizobium that nodulate soybean indicated two groups so distinct as to warrant classification as two species. Strains producing rhizobitoxine-induced chlorosis occur only in Group II, now classified as B. elkanii. Unlike B. japonicum, B. elkanii strains are characterized by (1) the ability to nodulate the rj1 genotype, (2) the formation of nodule-like structures on peanut, (3) a relatively high degree of ex planta nitrogenase activity, (4) distinct extracellular polysaccharide composition, (5) distinct fatty acid composition, (6) distinct antibiotic resistance profiles, and (7) low DNA homology with B. japonicum. Analysis with soybean lines near isogenic for the Rj4 versus rj4 alleles indicated that the Rj4 allele excludes a high proportion of B. elkanii strains and certain strains of B. japonicum such as strain USDA62 and three serogroup 123 strains. These groups, relatively inefficient in nitrogen fixation with soybean, tend to predominate in soybean nodules from many US soils. The Rj4 allele, the most common allelic form in the wild species, has a positive value for the host plants in protecting them from nodulation by rhizobia poorly adapted for symbiosis.
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  • 33
    ISSN: 1573-5036
    Keywords: herbaceous legumes ; moist savanna ; mycorrhiza ; nodulation ; nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The severity and increase of the Imperata cylindrica constraint as a weed, the decline of the traditional fallow systems as a means of soil fertility management and the lack of inorganic fertilizer appear to have created opportunities for adoption of mucuna (Mucuna pruriens) technology by smallholder farmers in some areas in the derived savanna of West Africa. What is not known, however, is the extent to which the establishment and N contribution of mucuna in these areas depend on symbiotic properties such as effective nodulation and mycorrhizal infection. Short term surveys carried out in 34 farmer's arable fields located in four different sites in the derived savanna, southern Benin, West Africa, together with results of greenhouse and field experiments showed that mycorrhizal infection rate of mucuma ranged from 2 to 31% and correlated positively with nodulation and shoot dry matter production. Nodulation occurred in 79% of the fields with numbers of nodules ranging from 0 to 135 plant−1. Mucuna responded both to inoculation and N fertilizer in degraded soils but growth response depended on the rhizobia strains and mucuna varieties. Mucuna accumulated in 12 weeks about 313 kg N ha−1 as either a sole crop or 166 kg N ha−1 when mixed/intercropped with maize, respectively. Across all cropping systems it derived an average of 70% of its N from atmospheric N2 (estimates made by the 15N isotope dilution method), representing 167 kg N ha−1 per 12 weeks in the field. Mucuna interplanted with maize obtained a greater proportion of its nitrogen (74%) from fixation than did mucuna grown alone (66%) suggesting that competition for soil N influences the proportion of nitrogen fixed by mucuna. The total amount of N2 fixed per hectare was, however, reduced significantly by intercropping mucuna with maize. A preceding mucuna crop provided a maize yield equivalent to 120 kg N kg ha−1 of inorganic N fertilizer.
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  • 34
    ISSN: 1573-5036
    Keywords: Bradyrhizobium japonicum ; genistein ; Glycine max ; grain yield ; nitrogen fixation ; protein yield ; short season
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In short-season soybean production areas, low soil temperature is the major factor limiting plant growth and yield. The decreases in soybean yield at low temperatures are mainly due to nitrogen limitation. Genistein, the most effective plant-to-bacterium signal in the soybean (Glycine max (L.) Merr.) nitrogen fixation symbiosis, was used to pretreat Bradyrhizobium japonicum. We have previously reported that this increased soybean nodulation and nitrogen fixation in growth chamber studies. Two field experiments were conducted on two adjacent sites in 1994 to determine whether the incubation of B. japonicum with genistein, prior to application as an inoculant, or genistein, without B. japonicum, applied onto seeds in the furrow at the time of planting, increased soybean grain yield and protein yield in short season areas. The results of these experiments indicated that genistein-preincubated bradyrhizobia increased the grain yield and protein yield of AC Bravor, the later maturing of the two cultivars tested. Genistein without B. japonicum, applied onto seeds in the furrow at the time of planting also increased both grain and protein yield by stimulation of native soil B. japonicum. Interactions existed between genistein application and soybean cultivars, and indicated that the cultivar with the greatest yield potential responded more to genistein addition.
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  • 35
    ISSN: 1573-5036
    Keywords: Acacia caven ; agroforestry ; Chamaecytisus proliferus subsp. palmensis ; isotope dilution ; 15N ; nitrogen fixation ; nodule efficiency ; Prosopis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Initial results of a long-term field experiment are presented for the above and below-ground biomass accumulation after two years, as well as root nodulation, nodule efficiency (g N fixed/g nodules) and biological N2 fixation (using the 15N isotope dilution method) of four N2-fixing tree species (NFTs) grown in the subhumid mediterranean-climate zone of central Chile. Two non-legume tree species, Fraxinus excelsior and Schinus polygamus, were used as reference plants for the isotope dilution calculations. Over two years, Tagasaste (Chamaecytisus proliferus subsp. palmensis, a Papilionoideae from the Canary Islands), produced 10 to 20 times more biomass than the other three NFTS (Acacia caven, Prosopis alba and P. chilensis); all Mimosoideae native to Chile, and nodulation and nitrogen fixed were an order of magnitude higher as well. At the end of the second year, the percentage of N derived from N2 fixation (%Ndfa) in Tagasate averaged 85.6, equivalent to ca. 49.1 g N fixed per tree. For all four NFTs, however, %Ndfa, nodule efficiency, and total N accumulation varied from one year to the next; caution is thus required in interpreting or predicting NFT performance over the long term, even if Tagasaste can already be considered a highly promising NFT for central Chile.
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  • 36
    ISSN: 1573-5036
    Keywords: inhibition ; 15N ; nitrate ; nitrogen fixation ; Trifolium repens ; uptake ; white clover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The impact of sustained low external concentrations of NO 3 − (0, 10, 100 and 1000 mmol m−3) on plant growth and the relative acquisition of N through N2 fixation and NO 3 − uptake by established, nodulated white clover (Trifolium repens L. cv. Blanca) was studied over 28 days in flowing solution culture. Nitrogen fixation was measured by N difference and 15N dilution methods. Plants supplied with NO 3 − achieved higher relative growth rates (% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGabmiEayaara% aaaa!3702!\[\bar x\]=0.091 d−1) compared with ‘control’ plants dependent on N2 fixation (0.073 d−1). Nitrate plants showed progressive increases in shoot: root d.w. ratios from 4 to 6.5–7.6 between days 0–28, compared with 5.1 on day 28 for control plants. Increases in both nodule d.w. and numbers per plant were inhibited after day seven at all concentrations of NO 3 − . The severity of inhibition of N2 fixation increased with increasing NO 3 − concentration and with time. The total amounts of N2 fixed per plant between days 0–7 after supplying 10, 100 and 1000 mmol m−3 NO 3 − , respectively, were 37–39, 28–30 and 0–13%, of the total N acquired. Between days 7–28 the proportional contributions of N2 fixation to total N acquisition declined to 3, 0.5 and 0%, respectively, in these treatments. The corresponding mean specific rates of N2 fixation between days 0–7 were, respectively, 5.4, 3.2, and 2.0 mmol N d−1 g−1 nodule d.w., compared with 7.9 mmol N d−1 g−1 nodule d.w. for zero NO 3 − plants. There was no evidence of a transitory increase in N2 fixation following the addition of NO 3 − , even at the lowest supply concentration.
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  • 37
    ISSN: 1573-5036
    Keywords: Bradyrhizobium ; hopanoids ; nitrogen fixation ; plant-microbe interaction ; Rhizobium ; squalene-hopene cyclase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Hopanoid lipids have been discovered recently in a number of nitrogen-fixing soil bacteria and in Bradyrhizobium bacteria which fix nitrogen in association with legume plants. We report here an investigation of the hopanoid content in an additional number of soil bacteria capable of living in close association with plants. Of the strains investigated, hopanoids were discovered in phototrophic, nitrogen-fixing bacteria and in an extended number of Bradyrhizobium strains. Strains in which hopanoids so far have not been found belong to the following genera: Rhizobium, Sinorhizobium, Phyllobacterium, Agrobacterium, and Azoarcus. To address the function of hopanoids in Bradyrhizobium, we cloned the gene coding for a key enzyme of hopanoid biosynthesis, the squalene-hopene cyclase, and expressed the gene in E. coli. The recombinant enzyme catalyzed in vitro the cyclization of squalene to hopanoid derivatives.
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  • 38
    ISSN: 1572-9680
    Keywords: nitrogen fixation ; 15N ; %Ndfa ; ryegrass+clover pasture ; silvopastoral system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Quantitative field measurements of biological nitrogen fixation (BNF) and biomass production by four different understorey pastures in a Pinus radiata-pasture agroforestry system were determined over a period of one year. The trees were two years old at the beginning of this study and the understorey pastures were being cut and removed for silage. The BNF was determined using the 15N dilution technique. Pastures of ryegrass+clover, cocksfoot+clover, phalaris+clover and lucerne were used. Substantial amounts of BNF were found (71 to 230 kg N ha−1 year−1) with lucerne showing the highest N fixation. However, lucerne derived only 71 to 72% of its N from the atmosphere (%Ndfa) during the spring/summer period compared to 83–97% with clovers, thus the net N demand from the soil was substantially higher with lucerne. This caused increased N stress to the trees. Clover in ryegrass+clover pasture fixed more N than the other grass+clover pastures. Although pasture position in relation to trees did not affect annual pasture total DMY and %Ndfa, pastures north of tree row grew better than those in other positions. Trees significantly affected the BNF of legumes and the botanical composition of pastures with highest BNF and legume production occurring in pastures midway between two rows of trees. These results suggest that it would be advantageous to evaluate different legumes and grasses for tolerance of shade and moisture stress in future studies. As the trees studied were only 1.5 to 3 m in height, their effects on BNF, seasonal pasture biomass production and botanical composition are expected to increase with tree dominance in the ecosystem with time. Amounts of N fixed were related to the productivity (i.e. dry matter and N yield) and seasonal persistence of the legumes. The productivity was high in spring and summer and low in autumn and winter.
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  • 39
    ISSN: 1573-5036
    Keywords: nitrogen ; nitrogen fixation ; stable isotopes ; tropical forest ; tropical pasture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The natural abundance of 15N was examined in soil profiles from forests and pastures of the Brazilian Amazon Basin to compare tropical forests on a variety of soil types and to investigate changes in the sources of nitrogen to soils following deforestation for cattle ranching. Six sites in the state of Rondônia, two sites in Pará and one in Amazonas were studied. All sites except one were chronosequences and contained native forest and one or more pastures ranging from 2 to 27 years old. Forest soil δ15N values to a depth of 1 m ranged from 8‰ to 23‰ and were higher than values typically found in temperate forests. A general pattern of increasing δ15N values with depth near the soil surface was broadly similar to patterns in other forests but a decrease in δ15N values in many forest profiles between 20 and 40 cm suggests that illuviation of 15N-depleted nitrate may influence total soil δ15N values in deeper soil where total N concentrations are low. In four chronosequences in Rondônia, the δ15N values of surface soil from pastures were lower than in the original forest and δ15N values were increasingly depleted in older pastures. Inputs of atmospheric N by dinitrogen fixation could be an important N source in these pastures. Other pastures in Amazonas and Pará and Rondônia showed no consistent change from forest values. The extent of fractionation that leads to 15N enrichment in soils was broadly similar over a wide range of soil textures and indicated that similar processes control N fractionation and loss under tropical forest over a broad geographic region. Forest δ15N profiles were consistent with conceptual models that explain enrichment of soil δ15N values by selective loss of 14N during nitrification and denitrification.
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  • 40
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    Nutrient cycling in agroecosystems 42 (1995), S. 241-250 
    ISSN: 1573-0867
    Keywords: Acetobacter diazotrophicus ; Azospirillum spp. ; cereals ; grasses ; Herbaspirillum spp. ; nitrogen fixation ; sugar cane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Over the last 20 years many new species of N2-fixing bacteria have been discovered in association with grasses, cereals and other non-nodulating crops. Virtually all of these bacteria are microaerophylic, fixing N2 only in the presence of low partial pressures of oxygen. Until a few years ago much attention was focussed on members the genusAzospirillum and it was assumed that N2 fixation was restricted to the rhizosphere or rhizoplane of the host plants. Through the use of N balance and15N techniques it has been shown that in the case of lowland rice, several tropical pasture grasses and especially sugar cane, the contributions of biological N2 fixation (BNF) are of agronomic significance. More detailed study of the N2-fixing bacteria associated with sugar cane (Acetobacter diazotrophicus andHerbaspirillum spp.) has shown that they occur in high numbers not only in roots of this crop but also in the stems, leaves and trash but are rarely found in the soil. Some of these endophytic diazotrophs have now also been found in forage grasses, cereals, sweet potato and cassava, although evidence of significant BNF contributions is still lacking. The identification of these endophytic diazotrophs as the organisms probably responsible for the high contributions of N2 fixation observed in sugar cane suggests that it may be possible to attain significant BNF contributions in some other gramineae and perhaps root crops.
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  • 41
    ISSN: 1573-5036
    Keywords: Acacia raddiana ; Acacia senegal ; Acacia seyal ; Faidherbia albida ; isotope dilution ; 15N ; nitrogen fixation ; Parkia biglobosa reference tree ; Tamarindus indica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A pot experiment was conducted in a greenhouse using the 15N isotope dilution method and two reference plants, Parkia biglobosa and Tamarindus indica to estimate nitrogen fixed in four Acacia species: A raddiana, A. senegal, A. seyal and Faidherbia albida (synonym Acacia albida). For the reference plants, the 15N enrichments in leaves, stems and roots were similar. With the fixing plants, leaves and stems had similar 15N enrichments; they were higher than the 15N enrichment of roots. The amounts of nitrogen fixed at 5 months after planting were similar using either reference plant. Estimates of the percentage of N derived from fixation (%Ndfa) for the above ground parts, in contrast to %Ndfa in roots, were similar to those for the whole plant. However, none of the individual plant parts estimated accurately total N fixed in the whole plant, and excluding the roots resulted in at least 30% underestimation of the amounts of N fixed. Between species, differences in N2 fixation were observed, both for %Ndfa and total N fixed. For %Ndfa, the best were A. seyal (average, 63%) and A. raddiana (average, 62%), being at least twice the %Ndfa in A. senegal and F. albida. Because of its very high N content, A. seyal was clearly the best in total N fixed, fixing 1.62 g N plant−1 compared to an average of 0.48 g N plant−1 for the other Acacia species. Our results show the wide variability existing between Acacia species in terms of both %Ndfa and total N fixed: A. seyal was classified as having a high N2 fixing potential (NFP) while the other Acacia species had a low NFP.
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  • 42
    ISSN: 1573-5036
    Keywords: colonization ; inoculation ; nitrogen fixation ; 15N and 33P dilution techniques ; plant-bacteria interaction ; phosphate-solubilizing bacteria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Pot experiments were carried out to investigate the effect of inoculation with pure and mixed cultures of nitrogen fixers Azospirillum lipoferum 137, Arthrobacter mysorens 7 and the phosphate-solubilizing strain Agrobacterium radiobacter 10 on growth and mineral nutrition of two barley cultivars. A significant positive effect on grain yield both of the studied barley cultivars was obtained after inoculation with mixtures of A. lipoferum 137 + A. radiobacter 10 and A. lipoferum 137 + A. mysorens 7 only. The acetylene reduction activity on roots or in batch culture was significantly higher when A. lipoferum 137 and A. radiobacter 10 were combined. Using 15N isotope dilution technique it was established that these mixed cultures significantly increased the accumulation of nitrogen fertilizer in the plants. The strain A. radiobacter 10 promoted a better accumulation of phosphorus fertilizer by plants and A. mysorens 7 increased the total phosphorus content in plant tissues. The maximum positive effect of joint inoculation on plant development was observed when the combined nitrogen in soil was in short supply. It was concluded that inoculation with bacterial mixtures provided a more balanced nutrition for the plants and the improvement in root uptake of nitrogen and phosphorus was the major mechanism of interaction between plants and bacteria. The introduced bacteria were able to colonize actively the rhizoplane of barley. No interspecific competition or antagonism were established between components of the bacterial mixtures in the rhizoplane. The strains A. mysorens 7 and A. radiobacter 10 improved viability of A. lipoferum 137 when the plants were grown in acid soil. Field experiments carried out on 3 barley cultivars confirmed the assertion that inoculation with mixed cultures significantly increases the grain yield and nitrogenous nutrition of plants as compared with single cultures.
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  • 43
    ISSN: 1573-5036
    Keywords: Glycine max ; nitrogen fixation ; nitrogen mobilization ; seed protein concentration ; soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Two high (NC106, NC111) and two normal (NC103, NC107) seed protein concentration lines, derived from two different recurrent selection populations of soybean (Glycine max L. Merr.) were subjected to partial defoliation at beginning seed fill (R5) under outdoor pot culture and field conditions. The aim of this study was to test the hypothesis that capacity to store N in vegetative organs and/or to mobilize that N to reproductive organs is associated with the high seed protein concentration trait. Symbiotic N2 fixation was the sole source of N in the pot experiment and the major source of N (met 〉 50% of the N requirement) in the low N soil used in the field experiment. Seed protein concentration and seed yield at maturity in both experiments and N accumulation and mobilization between R5 and maturity in the pot experiment were measured. The four genotypes did not differ significantly with respect to the amount of N accumulated before beginning seed fill (R5). Removal of up to two leaflets per trifoliolate leaf at R5 significantly decreased the seed protein concentration of NC107/111 but had no effect on this trait in NC103/106. Defoliation treatments significantly decreased seed yield, whole plant N accumulation (N2-fixation) during reproductive growth and vegetative N mobilization of all genotypes. Differences in harvest indices between the high and low protein lines accounted for approximately 35% of the differences in protein concentration. The two normal protein lines mobilized more vegetative N to the seed (average. 5.26 g plant−1) than the two high protein lines (average. 4.28 g plant−1). The two high seed protein lines (NC106, NC111) exhibited significantly different relative dependencies of reproductive N accumulation on vegetative N mobilization, 45% vs. 29%, in the control treatment. Whereas, NC103 with normal and NC106 with high seed protein concentration exhibited similar relative dependencies of reproductive N accumulation on vegetative N mobilization, (47% vs. 45%). Collectively, these results indicate that N stored in shoot organs before R5 and greater absolute and relative contribution of vegetative N mobilization to the reproductive N requirement are not responsible for the high seed protein concentration trait.
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  • 44
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    Plant and soil 175 (1995), S. 189-196 
    ISSN: 1573-5036
    Keywords: nitrogen fixation ; nitrogen-15 ; Sesbania rostrata ; Sesbania cannabina ; stem nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The flood adapted green manure legumes Sesbania cannabina and S. rostrata differ in their nodulation, the former nodulating on root only but the latter on both root and aerial stem. In the early wet season pre-rice niche in lowlands, these legumes are exposed to varying soil aeration and N status with NO3 - as the main N source in aerobic and NH4 + in flooded soil. To better use these legumes, the influence of soil aeration status and the resulting differences in soil N supply on N assimilation characteristics and biological nitrogen fixation (BNF) must be understood. We examined the interactions among soil aeration status, supply and form of mineral N, and BNF (estimated by 15N dilution) by growing S. rostrata and S. cannabina for 40 days in aerobic or flooded potted soil (3 kg) with zero to 3000 mg applied N. Both NH4 + and NO3 - were used in aerobic soil but only NH4 + in flooded soil. In aerobic soil, N accumulation potential of S. rostrata and S. cannabina were similar with NO3 -, NH4 +, or BNF as the major N source. Soil flooding increased N accumulation, but consistently more by S. rostrata than by S. cannabina. The maximum N accumulation of 275 mg plant-1 by S. rostrata was 20% greater than the maximum N accumulation by S. cannabina, in flooded soil, and 80% greater than maximum N accumulation by both legumes in aerobic soil. Regardless of soil aeration status and nodule location, mineral N increasingly substituted for biologically fixed N until total replacement occurred at non-limiting N supply. Reduction in the amount of BNF due to applied mineral N was greater in aerobic soil (with NO3 - as the N source) than in flooded soil (with NH4 + as the N source) for both legumes. Compared to root nodules, stem nodule BNF activity was less sensitive to NH4 + in flooded soils and resulted in more BNF-N in S. rostrata. Higher N requirement, stem-nodulation, and probably an ammoniphyllic character, allow S. rostrata to accumulate more N than S. cannabina in flooded soils.
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  • 45
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    Plant and soil 176 (1995), S. 161-169 
    ISSN: 1573-5036
    Keywords: Casuarina cunninghamiana ; Frankia ; nitrogen fixation ; nodulation ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A study was conducted to investigate the effects of phosphorus on nodule formation and function in the Casuarina-Frankia symbiosis. The effects of P on growth and survival of Frankia in the rhizosphere was assessed by examing Frankia growth and survival in flasks of basal nutrient solution. There was no growth in the nutrient solution during the experimental period. However, the viability of Frankia in the nutrient solution without P supply was half that of the initial level, whereas, with P supply, there was only a minor decline during the first week. In a growth pouch experiment, supplying P increased plant and nodule growth, irrespective of P status of the inoculant Frankia culture. There were no effects of P status on any growth or nodulation parameters measured when the inoculants had been standardized on the basis of viability. In a split root experiment, Frankia inoculation and application of P together or separately did not cause any significant difference. This suggests that growth and nodulation respond only to total P supply. Increasing P from 0.1 to 10 μM significantly increased plant growth but not N concentrations. Both nitrogen-fixation and nitrate supported growth were strongly increased as P increased from 0.1 to 1.0 μM. This study indicates that P deficiency limits the growth of host plants more severely than nitrogen fixation processes and P deficiency on nodulation and symbiotic nitrogen fixation in Casuarina cunninghamiana operated indirectly via reducing host plant growth.
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  • 46
    ISSN: 1573-5036
    Keywords: carbon balance ; carbon sink ; climate change ; grassland ; legume ; nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The response of plants to elevated CO2 is dependent on the availability of nutrients, especially nitrogen. It is generally accepted that an increase in the atmospheric CO2 concentration increases the C:N ratio of plant residues and exudates. This promotes temporary N-immobilization which might, in turn, reduce the availability of soil nitrogen. In addition, both a CO2 stimulated increase in plant growth (thus requiring more nitrogen) and an increased N demand for the decomposition of soil residues with a large C:N will result under elevated CO2 in a larger N-sink of the whole grassland ecosystem. One way to maintain the balance between the C and N cycles in elevated CO2 would be to increase N-import to the grassland ecosystem through symbiotic N2 fixation. Whether this might happen in the context of temperate ecosystems is discussed, by assessing the following hypothesis: i) symbiotic N2 fixation in legumes will be enhanced under elevated CO2, ii) this enhancement of N2 fixation will result in a larger N-input to the grassland ecosystem, and iii) a larger N-input will allow the sequestration of additional carbon, either above or below-ground, into the ecosystem. Data from long-term experiments with model grassland ecosystems, consisting of monocultures or mixtures of perennial ryegrass and white clover, grown under elevated CO2 under free-air or field-like conditions, supports the first two hypothesis, since: i) both the percentage and the amount of fixed N increases in white clover grown under elevated CO2, ii) the contribution of fixed N to the nitrogen nutrition of the mixed grass also increases in elevated CO2. Concerning the third hypothesis, an increased nitrogen input to the grassland ecosystem from N2 fixation usually promotes shoot growth (above-ground C storage) in elevated CO2. However, the consequences of this larger N input under elevated CO2 on the below-ground carbon fluxes are not fully understood. On one hand, the positive effect of elevated CO2 on the quantity of plant residues might be overwhelming and lead to an increased long-term below-ground C storage; on the other hand, the enhancement of the decomposition process by the N-rich legume material might favour carbon turn-over and, hence, limit the storage of below-ground carbon.
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  • 47
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    Plant and soil 170 (1995), S. 351-358 
    ISSN: 1573-5036
    Keywords: alfalfa ; cold soils ; inoculation ; Medicago ; nitrogen fixation ; rhizobia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Laboratory, growth chamber and field experiments were conducted to select among 226 isolates of Rhizobium meliloti for the ability to grow, nodulate alfalfa (Medicago sativa L.) and support N2-dependent plant growth between 9° and 12°C. There was wide variation in the abilities of R. meliloti isolates to grow and form nodules at 10°C. Culture doubling times (td) varied from 1 to 155h, and the number of nodules formed on alfalfa in growth pouches in 2 weeks varied from 0 to 3.8 nodules per plant. Nodulation occurred at 9°C, but there was no significant N2-dependent plant growth at this temperature. However, several isolates of R. meliloti had the ability to nodulate alfalfa and produce N2-dependent growth at root temperatures between 10° and 12°C root temperature than did 14 other isolates tested. In field experiments, inoculation with strain NRG-34 resulted in greater nodule numbers, nodule weight, proportion of nodules occupied by the inoculant strain and plant weight than did inoculation with a commercial strain (NRG-185). These results permitted selection of a strain with better low-temperature competitive abilities than the currently available commercial strains.
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  • 48
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    Plant and soil 174 (1995), S. 103-118 
    ISSN: 1573-5036
    Keywords: agropastoral systems ; farmer acceptability ; forage legume ; fertilization ; nitrogen fixation ; persistence ; tropical pastures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Forage legumes have long been lauded for their ability to fix atmospheric nitrogen and contribute to the sustainability of agricultural production systems. However despite the benefits they bring in terms of increased herbage and animal production they are not widely used in temperate or tropical regions. In this review the amounts of biological nitrogen fixation (BNF) needed to sustain the soil-plant-animal system are discussed and related to the amounts fixed in tropical pastures. The data suggest that tropical forage legumes have the capacity to meet the requirements to balance the N cycle of grazed pastures. The actual amounts required will depend on the rate of pasture utilization and the efficiency of recycling via litter, excreta and internal remobilization. The efficiency of nitrogen fixation (% of legume N derived from fixation) is usually high in tropical pastures (〉80%) and is unlikely to be affected by inorganic soil N in the absence of N fertilizer. Thus an estimate of the amoutns of N fixed could be obtained from simple estimates of legume biomass provided tissue levels of other nutrients such as phosphorus and potassium are adequate. Key factors for the achievement of sustainable grass/legume pastures include the selection of appropriate germplasm adapted to the particular environment and the judicious use of fertilizers such as phosphorus, potassium, calcium, magnesium and sulphur on acid infertile soils typical of the sub-humid and humid tropics. The main constraints to the widespread adoption of forage legumes include a lack of legume persistence, the presence of anti-quality factors such as tannins, variable Bradyrhizobium requirements and lack of acceptability by farmers. Strategies for the alleviation of these constrainst are discussed. Forage legumes can be used to recuperate degraded soils via their ability to improve the physical, chemical and biological properties of soils and these benefits could be of particular use for small-scale resource-poor farmers. The incorporation of forage legumes into agropastoral systems is discussed as an environmentally and economically attractive means to encourage the widespread adoption of legumes in the humid tropics.
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  • 49
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    Plant and soil 174 (1995), S. 279-286 
    ISSN: 1573-5036
    Keywords: chickpea ; genetics of BNF ; impact analysis ; nitrogen fixation ; nodulation variants ; Rhizobium inoculation ; technology adoption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Although viable Rhizobium inoculation technology for cultivated legumes has long been available, there has been little sustained adoption of this technology in tropical regions. Reasons contributing to this include inadequate demonstration of the technology, presence of adequate native rhizobia, high soil mineral nitrogen levels which suppress nitrogen fixation, inadequate quality control of Rhizobium inoculum and difficulties of inoculating under tropical conditions. In order to ensure a better adoption rate of existing or emerging biological nitrogen fixation (BNF) technologies, it is proposed that future research and development efforts better focus on the research-adoption-impact continuum. The salient features of this approach are described in this paper, using the example of recently developed nodulation variants in chickpea as a potential means of increasing BNF in this crop. It is suggested that previous experience with Rhizobium inoculation technology is amenable to ex-post impact analysis to analyze bottlenecks, and that ex-ante impact analysis should be built into on-going or planned BNF research, to better ensure that technology adoption occurs.
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  • 50
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    Plant and soil 171 (1995), S. 83-87 
    ISSN: 1573-5036
    Keywords: Al tolerance ; lime ; nitrogen fixation ; screening
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Two glasshouse experiments were conducted to refine screening procedures and to rank 36 native Acacia species for acid-soil tolerance. The first experiment showed that nitrogen fixation was more sensitive to soil acidity than plant growth per se. This suggests that, in screening experiments where acid soil tolerant rhizobia are not available, inorganic nitrogen should be supplied in determining the species' intrinsic ability to tolerate soil acidity and to avoid confounding sensitivity to acid soil with ineffectiveness of Rhizobium/Bradyrhizobium. The second experiment enabled us to rank 36 species into 4 classes of differing tolerance to acid soil. This experiment also indicated that the ranking order of species differed with the soil type, suggesting the need to use more than one soil type in screening experiments in order to obtain a better and more reliable ranking of species for tolerance to acid soil.
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  • 51
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    Plant and soil 173 (1995), S. 293-298 
    ISSN: 1573-5036
    Keywords: indole-3-acetic acid ; nitrogen fixation ; plant hormones ; Rhizobium ; tryptophan
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The effect that resistance to 5-methyltryptophan (MT) has on the symbiotic properties of B. japonicum was examined in a survey of fourteen clones. Resistance to MT often involves a mutational alteration in the regulation of tryptophan biosynthesis. Resistant clones (MTR) were isolated from agar plates containing MT. In the selection process care was taken to avoid pigmented clones that are likely to accumulate large amounts of indole compounds or show increased tryptophan catabolism. Wild-type control clones (WTc) were isolated from plates containing no selective agent. In greenhouse studies. Tracy-M soybean plants were inoculated with the two types of clones. After six weeks, plants which were inoculated with the MT resistant clones showed a much greater range of symbiotic effectiveness than did plants that received the control clones. While most MT-resistant clones were poor symbionts or unchanged in their symbiotic performance, one clone was obtained that had significantly improved symbiotic properties. The procedure may offer a way of selecting for clones with improved symbiotic performance. These results also indicate a link between tryptophan biosynthesis and symbiotic effectiveness.
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  • 52
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    Plant and soil 174 (1995), S. 181-194 
    ISSN: 1573-5036
    Keywords: green manure ; lowland rice ; nitrogen fertilizer ; nitrogen fixation ; Oryza sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The growing concern about the sustainability of tropical agricultural systems stands in striking contrast to a world-wide decline in the use of soil-improving legumes. It is timely to assess the future role that soil-improving legumes may play in agricultural systems. This paper reviews recent progress, potential, and limitations of green manure technology, using lowland rice cropping systems as the example. Only a few legume species are currently used as green manures in lowland rice. Sesbania cannabina is the most widely used pre-rice green manure for rice in the humid tropics of Africa and Asia. Astragalus sinicus is the prototype post-rice green manure species for the cool tropics. Stem-nodulating S. rostrata has been most prominent in recent research. Many green manure legumes show a high N accumulation (80–100 kg N ha-1 in 45–60 days of growth) of which the major portion (about 80%) is derived from biological N2 fixation. The average amounts of N accumulated by green manures can entirely substitute for mineral fertilizer N at current average application rates. With similar N use efficiencies, green manure N is less prone to loss mechanisms than mineral N fertilizers and may therefore contribute to long-term residual effects on soil productivity. Despite a high N2-fixing potential and positive effects on soil physical and chemical parameters, the use of green manure legumes for lowland rice production has declined dramatically world-wide over the last 30 years. Land scarcity due to increasing demographic pressure and a relatively low price of urea N are probably the main determining factors for the long-term reduction in pre-rice green manure use. Post-rice green manures were largely substituted for by high-yielding early-maturing grain legumes. Unreliability of green manure performance, non-availability of seeds, and labor intensive operations are the major agronomic constraints. The recognition and extrapolation of niches where green manures have a comparative advantage may improve an often unfavorable economic comparison of green manure with cash crop or fertilizer N. Socio-economic factors like the cost of land, labor, and mineral N fertilizer are seen to determine the cost-effectiveness and thereby farmers' adoption of sustainable pre-rice green manure technology. Hydrology and soil texture determine the agronomic competitiveness of a green manure with N fertilizers and with alternative cash crops. In general, the niches for pre-rice green manure are characterized by a relatively short time span available for green manure growth and a soil moisture regime that is unfavorable for cash crops (flood-prone rainfed lowlands with coarse-textured soils). Given the numerous agronomic and socio-economic constraints, green manure use is not seen to become a relevant feature of favourable rice-growing environments in the foreseeable future. However, in environments where soil properties and hydrology are marginal for food crop production, but which farmers may be compelled to cultivate in order to meet their subsistence food requirements, green manures may have a realistic and applicable potential.
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  • 53
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    Plant and soil 176 (1995), S. 189-196 
    ISSN: 1573-5036
    Keywords: Michaelis-Menten kinetics ; nitrogen fixation ; nitrate uptake ; Vicia faba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract An experiment was carried out to determine the relationship between nitrate uptake and nitrogen fixation of faba beans. Therefore inoculated and uninoculated faba beans were grown in nutrient solution with different nitrate concentrations. Nitrate uptake was measured every two days during the growing period. At the end of the experiment the nitrate uptake kinetics were determined with a short time depletion technique and nitrogen fixation was measured with the acetylene reduction method. A limitation of nitrate uptake due to nitrogen fixation was relatively small. Nitrate concentrations of approximately 1 mol m−3 and 5 mol m−3 decreased nitrogen fixation to values of 16% and 1% of the control plants which received no nitrate nitrogen. A reduction of nitrogen fixation was mainly due to a decrease of specific nitrogen fixation per unit nodule weight and to a lesser extent due to a reduction of nodule growth. Only the maximum nitrate influx (Imax) seemed to be influenced by nitrogen fixation. Michaelis-Menten constants (Km) and minimum NO inf3 − -concentrations (Cmin) were not significantly influenced by nitrogen fixation.
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  • 54
    ISSN: 1573-5060
    Keywords: acetylene reduction activity ; Bradyrhizobium japonicum ; genotype x strain interaction ; Glycine max ; soybean ; nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Bradyrhizobium japonicum strain G49 has been the only inoculum used in French soils. Soybean (Glycine max L. Merr.) cultivars were selected and tested according to their performances with this rhizobial strain. The aim of the present study was to determine the consequences of strain substitution on N2 fixation abilities of various genotypes. Three genotypes and cultivar Weber, in combination with B. japonicum strain G49 or SMGS1, were cultivated in pots and tested for nitrogenase activity under differing nitrogen nutrition conditions. The reliability of ARA (acetylene reduction activity) measurement for assessing symbiotic nitrogen fixation under the experimental conditions used was checked. Genotypic variability for symbiotic fixation activity was observed with each strain under soil culture conditions; important genotype x strain interactions were also involved. These results were corroborated for the protein yield and other yield component performances of the various genotype-strain associations. Thus, in France, the replacement of strain G49 with another one might result in the alteration of the relative agronomic performances of the soybean cultivars, since N2 fixation is considered as a major factor of soybean productivity.
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  • 55
    ISSN: 1573-5036
    Keywords: fast-growing rhizobia ; genetics ; nitrogen fixation ; soybean ; symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A soybean gene, Rfg1, controlling nodulation with strain USDA 205, the type strain for the fast-growing species Rhizobium fredii, was tested for allelism with the Rj4 gene. The Rj4 gene conditions ineffective nodulation primarily with certain strains of the slow-growing soybean microsymbiont, Bradyrhizobium elkanii. The F2 seeds of the cross of the cultivars Peking, carrying the alleles rfg1, Rj4, i (controlling inhibition of seed coat color) and W1 (controlling flower color), and Kent, carrying the alleles Rfg1, rj4, i-i and w1, were evaluated for nodulation response with strain USDA 205 by planting surface disinfested seeds in sterilized vermiculite in growth trays and inoculating with a stationary phase broth culture of strain USDA 205 at planting. Plants were classified for nodulation response visually after four weeks growth and transplanted to the field for F3 seed production. Flower color, purple (W1) vs white (w1), was determined in the field. The allele present at the i locus was determined by classification of F3 seed coat color. The F3 seeds were planted in growth trays and inoculated with strain USDA 61 of Bradyrhizobium elkanii to determine the genotype for the Rj4 locus. The Rfg1 and Rj4 genes were determined to be located at separate loci. Chi-square analysis for linkage indicated that Rfg1 segregated independently of the Rj4, I and W1 loci.
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  • 56
    ISSN: 1573-5036
    Keywords: actinorhizal plants ; cross-inoculation ; Frankia ; nitrogen fixation ; PCR ; 16S rRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Different Frankia strains and crushed nodule suspensions were tested for their ability to nodulate Coriaria nepalensis and Datisca cannabina. Datisca cannabina seedlings were nodulated effectively by both crushed nodule suspension from Coriaria nepalensis and Datisca cannabina. The origin of the endophyte in Datisca nodules induced by crushed nodules of Coriaria was confirmed by comparing partial PCR-amplified 16S rRNA sequences with those of the endophytes of both plants. Coriaria seedlings could only be nodulated by crushed nodule suspensions of Coriaria nepalensis. All pure cultures of Frankia used as a single inoculum source or in combinations with a nodule filtrate, failed to induce nodulation on Coriaria. Two atypical Frankia strains Cn3 and Cn7 isolated from Coriaria nodules showed no acetylene reduction activity and did not induce nodulation on the host seedlings.
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  • 57
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    Plant and soil 161 (1994), S. 135-145 
    ISSN: 1573-5036
    Keywords: actinorhizal ; Frankia ; nitrogen fixation ; root nodule ; symbioses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Over 200 species of angiosperms in eight different families are capable of forming root nodule symbioses with the actinomycetal genusFrankia as endosymbiont. Several thorough reviews of the biology of these actinorhizal associations have appeared in recent years (Benson and Silvester, 1993; Schwintzer and Tjepkema, 1990; Tjepkema et al., 1986). The purpose of the present discussion is to provide a summary overview of the actinorhizal symbioses, with an emphasis on recent research activities. A few areas of comparative interest with other symbiotic diazotrophs will be highlighted, especially regarding the question of oxygen protection and nitrogen fixation.
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  • 58
    ISSN: 1573-5036
    Keywords: inoculation ; Leucaena leucocephala ; nitrogen fixation ; nodulation ; persistence ; Rhizobium strains
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Establishment of Leucaena leucocephala was poor at Ibadan (Transition forest-savanna zone) and Fashola (savanna zone, 70 km north of Ibadan) in southwestern Nigeria as a result of low soil fertility and the presence of only a few native rhizobia capable of nodulating it. Inoculation with L. leucocephala at these two locations in 1982 resulted in striking responses with Rhizobium strains IRc 1045 and IRc 1050 isolated from L. leucocephala grown in Nigeria. The persistence of inoculated effective Rhizobium strains after inoculation is desirable since it removes the need for reinoculation. Because of the perennial nature of L. leucocephala and its use in long-term alley farming experiments, we examined the persistence of inoculated rhizobial strains after inoculation, and their ability to sustain N2-fixation and biomass production at Ibadan. In 1992, ten years after Rhizobium introduction, uninoculated, L. leucocephala fixed about 150 kg N ha-1 yr-1 or about 41% of total plant N compared to 180 kg N ha-1 yr-1 or 43% measured in 1982. Serological typing of the nodules using the Enzyme-Linked-Immunosorbent Assay (ELISA) and intrinsic resistance to the streptomycin test revealed that most of the nodules (96%) formed on L. leucocephala in 1992 were by Rhizobium strains IRc 1045 and IRc 1050, which were inoculated in 1982. Nodules were absent on uninoculated L. leucocephala grown on the adjacent field with no history of L. leucocephala cultivation. We conclude that the N2 fixed by Rhizobium strains IRc 1045 and IRc 1050 persisted for many years in the absence of L. leucocephala and sustained effectively fixed N2 which growth and yield of L. leucocephala after several years, thus encouraging a possible low-input alley farming system by smallholder farmers in Nigeria.
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  • 59
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    Plant and soil 158 (1994), S. 151-162 
    ISSN: 1573-5036
    Keywords: acetylene reduction ; gas exchange ; hydrogen evolution ; nitrogen fixation ; oxygen diffusion ; rhizobia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The closed acetylene reduction assay has been used as a measure of nitrogenase activity and an indicator of N2 fixation in Rhizobium/legume symbioses for 25 years. However, starting 10 years ago this assay has come under harsh criticism as being inaccurate. Currently, confusion exists regarding the conditions under which the acetylene reduction assay can be used accurately, or whether it can be used at all as a measure of nitrogenase activity. This article reviews the circumstance that has lead to this confusion. The author argues that under the proper assay conditions and with the appropriate checks, the closed acetylene reduction assay is still a valuable tool in assessing relative differences in nitrogenase activity in Rhizobium/legume symbioses.
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  • 60
    ISSN: 1573-5036
    Keywords: acetylene reduction ; errors ; gas exchange ; nitrogen fixation ; nodules ; rhizobia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract This article is in response to that of Vessey (1994) who argues that the traditional, closed acetylene reduction assay can still be a valuable tool for measuring relative differences in nitrogenase activity of legumes. To counter this assertion we consider the practical uses of the traditional assay procedure in relation to real research situations. This requires the use of the assay to be considered separately in the different circumstances of pot-grown and field-grown plants. We conclude that for pot-grown legumes there are a few practical applications where the use of the traditional, closed assay procedure is valid and we accept that these can be extended by the careful use of calibrations against open, flow-through systems. However, we doubt that there are many situations where such a calibration approach would have practical advantages over using the flow-through system to obtain the actual measurements. We cannot recommend any form of the uncalibrated acetylene reduction assay for field-based studies and suggest that researchers consider the merits of simple, alternative measurements such as dry weight, yield and total nitrogen.
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  • 61
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    Plant and soil 159 (1994), S. 233-243 
    ISSN: 1573-5036
    Keywords: alfalfa ; Medicago sativa ; nitrogen excretion ; 15N-labelled gas ; nitrogen fixation ; nitrogen transfer ; rhizosphere soil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Rhizodeposition has been proposed as one mechanism for the accumulation of significant amounts of N in soil during legume growth. The objective of this experiment was to directly quantify losses of symbiotically fixed N from living alfalfa (Medicago sativa L.) roots to the rhizosphere. We used 15N-labeled N2 gas to tag recently fixed N in three alfalfa lines [cv. Saranac, Ineffective Saranac (an ineffectively nodulated line), and an unnamed line in early stages of selection for apparent N excretion] growing in 1-m long polyvinylchloride drainage lysimeters in loamy sand soil in a greenhouse. Plants were in the late vegetative to flowering growth stage during the 2-day labelling period. We determined the fate of this fixed N in various plant organs and soil after a short equilibration period (2 to 4 days) and after one regrowth period (35 to 37 days). Extrapolated N2 fixation rates (46 to 77μg plant−1 h−1) were similar to rates others have measured in the field. Although there was significant accretion of total N in rhizosphere compared to bulk soil, less than 1% was derived from newly fixed N and there were no differences between the ‘excreting’ line and Saranac. Loss of N in percolate water was small. These results provide the first direct evidence that little net loss of symbiotically-fixed N occurs from living alfalfa roots into surrounding soil. In addition, these results confirm our earlier findings, which depended on indirect 15N labelling techniques. Net N accumulation in soil during alfalfa growth is likely due to other processes, such as decomposition of roots, nodules, and above ground litter, rather than to N excretion from living roots and nodules.
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  • 62
    ISSN: 1573-5036
    Keywords: Azospirillum ; Azotobacter ; Klebsiella ; nitrogen fixation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A plentiful supply of fixed nitrogen as ammonium (or other compounds such as nitrate or amino acids) inhibits nitrogen fixation in free-living bacteria by preventing nitrogenase synthesis and/or activity. Ammonium and nitrate have variable effects on the ability ofRhizobiaceae (Rhizobium, Bradyrhizobium andAzorhizobium) species to nodulate legume hosts and on nitrogen fixation capacity in bacteroid cells contained in nodules or in plant-free bacterial cultures. In addition to effects on nitrogen fixation, excess ammonium can inhibit activity or expression of other pathways for utilization of nitrogenous compounds such as nitrate (through nitrate and nitrite reductase), or glutamine synthetase (GS) for assimilation of ammonium. This paper describes the roles of two key genesglnB andglnD, whose gene products sense levels of fixed nitrogen and initiate a cascade of reactions in response to nitrogen status. While work onEscherichia coli and other enteric bacteria provides the model system,glnB and, to a lesser extent,glnD have been studied in several nitrogen fixing bacteria. Such reports will be reviewed here. Recent results on the identity and function of theglnB andglnD gene products inAzotobacter vinelandii (a free-living soil diazotroph) and inRhizobium leguminosarum biovarviciae, hereinafter designatedR.l. viciae will be presented. New data suggests thatAzotobacter vinelandii probably contains aglnB-like gene and this organism may have twoglnD-like genes (one of which was recently identified and namednfrX). In addition, evidence for uridylylation of theglnB gene product (the PII protein) ofR. l. viciae in response to fixed nitrogen deficiency is presented. Also, aglnB mutant ofR. l. viciae has been isolated; its characteristics with respect to expression of nitrogen regulated genes is described.
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  • 63
    ISSN: 1573-5036
    Keywords: Frankia ; nitrogen fixation ; symbioses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Patterns of nodulation, growth, andFrankia — host specificity have not been well characterized for the actinorhizal genera in the family Rosaceae because of the scarcity ofFrankia isolates from these taxa. Furthermore, the few isolates available from actinorhizal Rosaceae have consistently failed to nodulate plants from the host genus. In a series of experiments, species of rosaceousDryas, Cowania, Cercocarpus, Fallugia, andPurshia were inoculated withFrankia isolates, crushedDryas actinorhizae, and neoglacial soils to ascertain whether any of these inocula would effectively induce nodulation. Neoglacial soils from Alaska and Canada nodulated not only the localDryas drummondii, but alsoCercocarpus betuloides, Cowania mexicana andPurshia tridentata from distant and ecologically diverse locales as well as nonrosaceous, actinorhizal species ofAlnus, Elaeagnus, Myrica, andShepherdia. But of eightFrankia isolates, including two fromPurshia tridentata and one fromCowania mexicana, none were able to induce nodulation onPurshia orCowania species. Globular, actinorhizae-like nodules incapable of acetylene reduction were produced onC. betuloides inoculated withFrankia isolates. Crushed nodule suspensions fromDryas drummondii nodulated rosaceousCowania, Dryas andPurshia, as well as non-rosaceousElaeagnus, Myrica, andShepherdia species. Nodules produced by inoculation ofCowania mexicana andPurshia tridentata with crushed, dried nodule suspensions fromDryas drummondii reduced acetylene to ethylene, indicating nitrogenase activity for these nodulated plants. These data suggest that a similar microsymbiont infects the actinorhizal genera in the family Rosaceae.
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  • 64
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    Plant and soil 163 (1994), S. 157-164 
    ISSN: 1573-5036
    Keywords: nitrogen fixation ; nodule occupancy ; Phaseolus vulgaris ; Rhizobium leguminosarum bv.phaseoli ; soil acidity ; strain competition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The effect of soil pH on the competitive abilities of twoRhizobium leuminosarum bv.phaseoli type I and one type II strains was examined in a nonsterile soil system.Phaseolus vulgaris seedlings, grown in unlimed (pH 5.2) or limed (pH 7.6) soil, were inoculated with a single-strain inoculum containing 1 × 106 cells mL−1 of one of the three test strains or with a mixed inoculum (1:1, type I vs. type II) containing the type II strain CIAT 899 plus one type I strain (TAL 182 or CIAT 895). At harvest, nodule occupants were determined. In a separate experiment, a mixed suspension (1:1, type I vs. type II) of CIAT 899 paired with either TAL 182 or CIAT 895 was used to inoculateP. vulgaris seedlings grown in sterile, limed or unlimed soil. The numbers of each strain in the rhizosphere were monitored for 10 days following inoculation. The majority of nodules (〉 60%) formed on plants grown in acidic soil were occupied by CIAT 899, the type II strain. This pattern of nodule occupancy changed in limed soil. When CIAT 899 was paired with TAL 182, the type I strain formed 78% of the nodules. The number of nodules formed by CIAT 899 and CIAT 895 (56% and 44%, respectively) were not significantly different. The observed patterns of nodule occupancy were not related to the relative numbers or specific growth rates of competing strains in the host rhizosphere prior to nodulation. The results indicate that soil pH can influence which symbiotype ofR. leguminosarum bv.phaseoli will competitively nodulateP. vulgaris.
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  • 65
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    Plant and soil 158 (1994), S. 135-139 
    ISSN: 1573-5036
    Keywords: clover ; inoculation ; nitrogen fixation ; nodulation ; rhizobia ; Trifolium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The relationship between numbers of rhizobia and nodulation response of legumes is of considerable practical importance. Experiments were done under controlled conditions to determine the influence of numbers of Rhizobium leguminosarum biovar. trifolii on nodulation of arrowleaf clover (Trifolium vesiculosum Savi.) and crimson clover (T. incarnatum L.). Numbers of rhizobia in excess of 1000 per seed did not substantially increase earliness of nodulation or total number of nodules formed on the taproot. Nodules, however, were formed nearer the top of the taproot as numbers of rhizobia increased to 100,000 per seed. Delayed inoculation experiments indicated that nodulation sites for these clovers only remained susceptible to infection for less than 1 day. Delaying inoculation for 4 days resulted in only a 1 to 2 day delay in nodulation for arrowleaf and crimson clovers respectively and no delay for subterranean clover (T. subterraneum L.). Apparently, larger seedlings nodulated faster.
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  • 66
    ISSN: 1573-5036
    Keywords: allelopathy ; Carduus nutans L. ; competition ; nitrogen fixation ; plant invasions ; Trifolium repens L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Carduus nutans L. is an invasive pasture/grassland species which may undergo rapid population growth through positive feedback. Plants ofC. nutans produce a vegetative rosette, and after several months produce stems containing flower-heads, during which time the rosette leaves die and decompose. We investigated the influence ofC. nutans on the nitrogen-fixation ability ofTrifolium repens L. in three experiments. The first experiment was set up in a “mixture” design, and demonstrated that seedlings ofT. repens were more susceptible to competition with otherT. repens seedlings than toC. nutans seedlings. Nodule numbers and acetylene reduction per unit root, and acetylene reduction per unit nodules were adversely affected by increasingT. repens, but notC. nutans densities. The second experiment was of an additive design, with separate partitions to isolate above-ground and below-ground interference. FloweringC. nutans plants strongly inhibitedT. repens root growth, nodulation and acetylene reduction, but usually only when shoot interference was permitted. This appears to be due to decomposition of rosette leaves, which was maximal at this stage. The third experiment involved monitoring effects of taggedC. nutans individuals againstT. repens in the field. This experiment showed that acetylene reduction was severely influenced by floweringC. nutans (when rosette leaves were decomposing), even when only mild reduction ofT. repens growth was observed, and these effects persisted for some months after theC. nutans plants had died. The results of these experiments in combination suggest that decomposing rosette leaves have a strong potential to inhibitT. repens nitrogen fixation. It appears that allelopathy is involved, since alternative explanations (e.g. root competition byC. nutans; effects ofC. nutans on soil moisture, microbial nutrient immobilisation and light availability; facilitation of herbivores byC. nutans) can be effectively discounted. Although invasive species are often assumed to be associated with soil nitrogen build-up, we believe that some invasive species such asC. nutans have the potential to induce long-term decline of soil nitrogen input.
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  • 67
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    Plant and soil 165 (1994), S. 55-65 
    ISSN: 1573-5036
    Keywords: CO2 enrichment ; decomposition ; mycorrhizae ; nitrogen fixation ; rhizosphere ; soil biota ; soil fauna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Summary Predictably, the responses of soil biota to CO2 enrichment and the degree of experimental emphasis on them increase with proximity to, and intimacy with, roots. Symbiotic associations are all stimulated to some degree. Total plant mycorrhization increases with elevated CO2. VAM fungi increase proportionately with fine root length/mass increase. ECM fungi, however, exhibit greater colonization per unit root length/mass at elevated CO2 than at current atmospheric levels. Total N-fixation per plant increases in all species examined, although the mechanisms of increase, as well as the eventual benefit to the host relative to N uptake may vary. Microbial responses are unclear. The assumption that changes in root exudation will drive increased mineralization and facilitate nutrient uptake should be examined experimentally, in light of recent models. Microbial results to date suggest that metabolic activity (measured as changes in process rates) is stimulated by root C input, rather than population size (measured by cell or colony counts). Insufficient evidence exists to predict responses of either soil-borne plant pathogens or soil fauna (i.e., food web responses). These are areas requiring attention, the first for its potential to limit ecosystem production through disease and the second because of its importance to nutrient cycling processes. Preliminary data on foliar litter decomposition suggests that neither nutrient ratios nor decomposition rates will be affected by rising CO2. This is another important area that may be better understood as the number of longer term studies with more realistic CO2 exposures increase. Evidence continues to mount that C fixation increases with CO2 enrichment and that the bulk of this C enters the belowground component of ecosystems. The global fate and effects of this additional C may affect all hierarchical levels, from organisms to ecosystems, and will be largely determined by responses of soil biota.
    Notes: Abstract Increasing concentrations of atmospheric CO2 could have dramatic effects upon terrestrial ecosystems including changes in ecosystem structure, nutrient cycling rates, net primary production, C source-sink relationships and successional patterns. All of these potential changes will be constrained to some degree by below ground processes and mediated by responses of soil biota to indirect effects of CO2 enrichment. A review of our current state of knowledge regarding responses of soil biota is presented, covering responses of mycorrhizae, N-fixing bacteria and actinomycetes, soil microbiota, plant pathogens, and soil fauna. Emphasis will be placed on consequences to biota of increasing C input through the rhizosphere and resulting feedbacks to above ground systems. Rising CO2 may also result in altered nutrient concentrations of plant litter, potentially changing decomposition rates through indirect effects upon decomposer communities. Thus, this review will also cover current information on decomposition of litter produced at elevated CO2.
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  • 68
    ISSN: 1573-5060
    Keywords: red clover ; strains ; isotope dilution ; nitrogen fertilizer ; nitrogen fixation ; Trifolium pratense
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Genetic variation in fixed nitrogen (N) yield of red clover (Trifolium pratense L.) strains and cultivars was investigated using the 15N isotope dilution method under three regimes of N fertilization: 0.5, 30, and 60 N (kg N ha−1 per cut). The yield of fixed N per cut (the mean of eight cuts over 2 production years) varied among the strains (progenies of crosses between inbred parents) from 148 to 443 mg per plant at 0.5 N, from 76 to 324 mg at 30 N, and from 69 to 300 mg at 60 N. There were significant and consistent strain differences in the percentage of clover N derived from the atmosphere (% Ndfa). However, %Ndfa was positively correlated with dry mass yield. Consequently, ranking of the strains according to fixed N yield reflected that of dry mass yield. There were only minor strain × N fertilizer interactions, suggesting that selection for enhanced N fixation can be carried out at a single rate of fertilizer N. For a selected pair of strains, the difference in yield of fixed N was confirmed in an Italian ryegrass-red clover mixture, both without and with the addition of N fertilizer (50 kg N ha−1 per cut). Results with 7-week-old seedling plants in a growth chamber, although obtained in the presence of mineral N and with the isotope dilution method, did not adequately predict field performance. It is concluded that selection for dry matter or total N yield is likely to result in an enhanced yield of fixed N at any level of mineral N availability.
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  • 69
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    Plant and soil 161 (1994), S. 109-114 
    ISSN: 1573-5036
    Keywords: defoliation ; nitrogen fixation ; nitrogen sink-strength ; nitrogenase activity ; oxygen diffusion ; white clover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The dramatic decrease in nitrogenase activity after the defoliation of forage legumes has been recognized for a long time; however, the underlying mechanisms are not understood yet. The impact of current photosynthesis can be excluded. The precise role of carbohydrate availability is still unclear and remains to be established. From current knowledge we can conclude that, after defoliation, nitrogenase activity in legume nodules is down-regulated by a variable oxygen diffusion resistance. The triggering elements are not known; there is, however, increasing evidence that the plant's demand for symbiotically fixed nitrogen plays an important role. The possibility is here discussed that, after defoliation, a nitrogen feedback mechanism regulates nitrogenase activity through a variable oxygen diffusion resistance in the nodules.
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  • 70
    ISSN: 1573-5036
    Keywords: actinorhizal ; Alnus glutinosa ; acetylene reduction ; carbon dioxide ; Frankia ; nitrogen fixation ; nodules ; roots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The effects of aeration of the N-free rooting medium with elevated CO2 on (a) acetylene reduction by perlite-grown plants and (b) N2-fixation and long-term growth of nutrient solution-grown plants were determined for nodulatedAlnus glutinosa (L.) Gaertn. In the former experiments, roots of intact plants were incubated in acetylene in air in darkened glass jars for 3 hr, followed by a further 3 hr incubation period in air enriched with CO2 (0–5%). During incubation, the CO2 content of the jars increased by 0.17% per hour due to respiration of the root system, so that the CO2 content at 3 hr was 0.5%. Additional enrichment of the rooting medium gas-phase with CO2 equivalent to 1.1% and 1.75% CO2 of the gas volume significantly increased nitrogenase activity (ethylene production) by 55% and 50% respectively, while enrichment with greater than 2.5% CO2 decreased activity. In contrast, ethylene production by control plants, where CO2 was not added to the assay jars, decreased by 8% over the assay period. In long-term growth experiments, nodulated roots of intactAlnus glutinosa plants were sealed into jars containing N-free nutrient solution (pH 6.3) and aerated with air, or air containing elevated levels of CO2 (1.5% and 5%). Comparison of the appearance of CO2-treated with air treated plants suggested that 1.5% CO2 stimulated plant growth. However, at harvest after 5 or 6 weeks variability between plants masked the significance of differences in plant dry weight. A significant increase of 33% in total nitrogen of plants aerated with 1.5% CO2, compared with air-treated plants, was demonstrated, broadly in line with the short-term increase in acetylene reducing activity observed following incubations with similar CO2 concentrations. Shoot dry weight was not affected significantly by long-term exposure to 5% CO2, the main effect on growth being a 20% reduction in dry weight of the root system, possibly through inhibition of root system respiration. However, in contrast to the inhibitory effects of high CO2 on acetylene reduction there was no significant effect on the amounts of N2 fixed.
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  • 71
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    Plant and soil 152 (1993), S. 19-23 
    ISSN: 1573-5036
    Keywords: acetylene reduction ; difference method ; legumes ; 15N isotope method ; nitrogen ; nitrogen fixation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract To assure proper management and fully realize the benefits of the legume-Rhizobium symbiosis it is necessary to be able to quantify the amount of nitrogen fixed. Having measured the effectiveness of atmospheric N2 fixation the macro- or micro-symbionts as well as agronomic factors can be manipulated with the objective to maximize biological nitrogen fixation. A suitable method to quantify nitrogen fixation is therefore necessary in any programme aiming at increasing N2 fixation, like the one being reported in this volume. There are several methods available to quantify nitrogen fixation and most of the commonly used ones are described in the present paper listing their advantages and disadvantages.
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  • 72
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    Plant and soil 155-156 (1993), S. 349-352 
    ISSN: 1573-5036
    Keywords: copper nutrition ; iron ; leghaemoglobin ; Lupinus luteus L. ; nitrogen fixation ; polyphenol oxidase activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The effect of copper nutrition on symbiotic N2 fixation in Lupinus luteus L. was studied. Copper nutrition increased the yield, total nitrogen content and dry weight of nodules. The control plants did not produce pods. Copper deficiency limited iron uptake and its translocation to the nodules. Nodules of copper-deficient plants contained less than half the leghaemoglobin concentration of copper-adequate plants and about one third the polyphenol oxidase activity, tested with catechol as a substrate.
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  • 73
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    Plant and soil 149 (1993), S. 103-109 
    ISSN: 1573-5036
    Keywords: common beans ; gliricidia ; leucaena ; lonchocarpus ; nitrogenase ; nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Common bean (Phaseolus vulgaris L.) represents an important crop in tropics, but previous screenings of Rhizobium leguminosarum bv. phaseoli did not show strains that could fix N2 in symbiosis with bean at temperatures higher than 35°C (Hungria and Franco, 1993). However, there are other rhizobia and bradyrhizobia species that nodulate some tropical leguminous trees and can fix N2 at high temperatures. In a trial of rhizobial strains isolated from leguminous trees, we found that 14 out of 21 isolates from Gliricidia, Lonchocarpus and Leucaena were also able to nodulate common beans at optimal temperatures (28/23°C, day/night). When we exposed beans inoculated with these strains to high temperature conditions, 40°C/8 h/day, some of them accumulated at flowering time as much or more N as bean plants receiving mineral N. These broad host-range sources of rhizobia capable of fixing nitrogen with bean at high temperature seem to have the potential to improve yields in tropical soils.
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  • 74
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    Keywords: carbohydrates ; leghemoglobin ; nitrogen fixation ; nitrogenase ; ontogeny ; Pisum sativum ; pod-filling ; protein ; proteolytic activity ; respiration ; Rhizobium leguminosarum ; root nodules ; starch
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Root nodule ontogeny was followed in different parts of the root system of field peas (Pisum sativum L. cv. Century) to investigate the contribution to total nitrogen fixation by different nodule subpopulations. Seed-inoculated plants were grown to maturity in controlled-environment growth chambers. In a flow-through system nitrogenase activity (H2-evolution in air) and nodulated-root respiration (net CO2-evolution) were measured weekly or biweekly in different parts (top and mid) of the root system. Root nodule extracts were assayed for total soluble cytosolic protein, total heme, proteolytic capacity (at pH 7.0), soluble carbohydrates and starch. Total nitrogenase activity and nodule respiration were higher in the top zone, which was explained by differences in root and nodule mass. Nodule specific nitrogenase activity was similar in both zones, and gradually declined throughout the experiment. No differences were found between nodule subpopulations in the dry-matter specific concentrations of glucose, fructose, sucrose or starch. Neither did nodule concentrations of protein or leghemoglobin differ between the zones. Throughout reproductive growth, no decline was found in total or nodule specific nitrogenase activity, in any of the nodule subpopulations. Growth of the root nodules continued throughout the experiment, though growth of shoot and roots had ceased. The data gives no support for carbohydrate limitation in root nodules during pod-filling, since nodule respiration remained high, the concentration of soluble carbohydrates increased significantly, and the amount of starch was not reduced. We conclude that when this symbiosis is grown under controlled conditions, nitrogenase activity in nodules sub-sampled from the crown part of the root system is representative for the whole nodule population.
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  • 75
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    Plant and soil 151 (1993), S. 167-174 
    ISSN: 1573-5036
    Keywords: glutamine synthetase ; glutamate synthase ; nitrogen fixation ; plant genotype ; root nodules ; Vicia faba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Five inbred lines and a commercial cultivar of field-bean (Vicia faba) were inoculated with a strain of Rhizobium leguminosarum bv. viciae and grown, with or without nitrate, under axenic conditions in a growth chamber. There were significant (p〈0.01) differences between lines in N yield, both in N-free solution (N2 fixed, fully symbiotic) and with added NO3 −. Increase in the N yield of NO3 −-fed plants compared with fully symbiotic plants also varied between genotypes, with line VF109 being the least responsive to NO3 −. This may indicate nitrate tolerance in relation to N2 fixation. The indication was supported by the observations that nodule development and nodule activity (acetylene reduction) were less inhibited in VF109 in the presence of 8 mM NO3 − than in the other lines of Vicia faba. Glutamine synthetase and glutamate synthase (GOGAT) activities appeared to be related to genotypic differences in symbiotic efficiency. This finding suggests that assays of nodule glutamine synthetase and NADH-glutamate synthase might be used in Vicia faba breeding programs to indicate capacity for symbiotic N2 fixation, particularly in the presence of NO3 −. ei]Section editor: B G Rolfe
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  • 76
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    Plant and soil 152 (1993), S. 71-79 
    ISSN: 1573-5036
    Keywords: common bean ; nitrogen fixation ; Phaseolus vulgaris L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Common bean (Phaseolus vulgaris L.), which is an important food crop in the Americas, Africa and Asia, usually is thought to fix only small amounts of atmospheric nitrogen. However, field data indicate considerable genetic variability for total N2 fixation and traits associated with fixation. Studies have shown that selection to increase N2 fixation will be successful if: (1) discriminating traits (selection criteria) are measured precisely, (2) variability in germplasm is heritable, (3) selected parents are also agronomically suitable, (4) units of selection facilitate quantification of selection criteria, and (5) a breeding procedure that allows maximum genetic gain for N2 fixation and recombination with essential agronomic traits is chosen. Breeding lines capable of fixing enough atmospheric N2 to support seed yields of 1000–2000 kg ha−1 have been identified and new cultivars with high N2 fixation potential are being released.
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  • 77
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    Plant and soil 152 (1993), S. 87-91 
    ISSN: 1573-5036
    Keywords: common bean ; 15N isotope dilution ; nitrogen fixation ; Phaseolus vulgaris L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Two field experiments were performed to evaluate the nitrogen fixation potential of twenty common bean cultivars and breeding lines during summer and winter seasons of 1986 and 1988, respectively. The 15N isotope dilution method was used to quantify N2 fixation. The cultivars and breeding lines were variable in terms of their N2 fixation. The cv. Caballero was very efficient, with more than 50% N derived from the atmosphere and 60–80 kg N ha−1 fixed in both seasons. Other cultivars were less efficient, since the poorest ones derived less than 30% of their nitrogen from the atmosphere and fixed less than 20 kg N ha−1. After additional testing the best cultivars may be used directly by the farmers for cultivation. The experiments have provided information about which genotypes may be used to breed for enhanced fixation in common bean.
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  • 78
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    Plant and soil 152 (1993), S. 115-121 
    ISSN: 1573-5036
    Keywords: common bean ; nitrogen assimilation ; nitrogen fixation ; nitrogen partitioning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Field and greenhouse experiments were conducted to assess the nitrogen fixation rates of four cultivars of common bean (Phaseolus vulgaris L.) at different growth stages. The 15N isotope dilution technique was used to quantify biological nitrogen fixation. In the greenhouse, cultivars M4403 and Kallmet accumulated 301 and 189 mg N plant−1, respectively, up to 63 days after planting (DAP) of which 57 and 43% was derived from atmosphere. Under field conditions, cultivars Bayocel and Flor de Mayo RMC accumulated in 77 DAP, 147 and 135 kg N ha−1, respectively, of which approximately one-half was derived from the atmosphere. The rates of N2 fixation determined at different growth stages increased as the plants developed, and reached a maximum during the reproductive stage both under field and greenhouse conditions. Differences in translocation of N were observed between the cultivars tested, particularly under field conditions. Thus, the fixed N harvest index was 93 and 60 for cultivars Flor de Mayo and Bayocel, respectively. In early stages of growth, the total content of ureides in the plants correlated with the N fixation rates. The findings reported in the present paper can be used to build a strategy for enhancing biological N2 fixation in common bean.
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  • 79
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    Keywords: Actinorhiza ; Betula pendula ; Betula pubescens ; birch ; Enterobacter agglomerans ; Festuca rubra ; Frankia ; Klebsiella pneumoniae ; nitrogen fixation ; Poa pratensis ; Pseudomonas sp. ; rhizosphere ; root
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Bacterial growth in the rhizosphere and resulting changes in plant growth parameters were studied in small aseptic seedlings of birch (Betula pendula and B. pubescens) and grasses (Poa pratensis and Festuca rubra). The seedlings were inoculated with three Frankia strains (Ai1a and Ag5b isolated from native Alnus root nodules and Ai17 from a root nodule induced by soil originating from a Betula pendula stand), and three associative N2-fixing bacteria (Enterobacter agglomerans, Klebsiella pneumoniae and Pseudomonas sp., isolated from grass roots). Microscopic observations showed that all the Frankia strains were able to colonize and grow on the root surface of the plants tested without addition of an exogenous carbon source. No net growth of the associative N2-fixers was observed in the rhizosphere, although inoculum viable counts were maintained over the experimental period. Changes in both the biomass and morphology of plant seedlings in response to bacterial inoculation were recorded, which were more dependent on the plant species than on the bacterial strain.
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  • 80
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    Plant and soil 155-156 (1993), S. 353-354 
    ISSN: 1573-5036
    Keywords: cowpea ; nitrogen fixation ; nitrate ; partitioning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract If the quality and quantity of yields from cowpea (Vigna unguiculata [L.] Walp.) are to be maximised, a complete understanding of the N nutrition of the plant must be achieved. The N requirement for developing pods of this species may come from mobilization of N in vegetative tissue, biological N fixation and uptake of N from soil. In this study, the fate of a pulse of fixed 15N2 or of 15NO3-given to different cowpea plants during pod development was determined. The plants were grown in vermiculite in plastic pots that were able to be sealed with silicone adhesive and equipped with a rubber septum so that 15N2 gas could be injected into the air space above the vermiculite, and gas losses would be eliminated. Nineteen days after injection of 15N2 the pods, leaves, nodules and roots contained 65%, 15%, 9%, and 4%, respectively of the quantity of 15N2 fixed. When 15NO3-15N was taken up by other plants during this period, these plant parts contained 40%, 26%, 3% and 19%, respectively, of the total plant 15N. The percentage 15N in roots was greater, and that of 15N in nodules was lower, when 15NO3-15N was applied than when 15N2 was utilised by plants. These results indicate that, while a high percentage of fixed-N or NO3-N given to cowpea plants moved to the developing pods, other sinks were competing for this newly-aquired N.
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  • 81
    ISSN: 1573-5036
    Keywords: mutant ; 15N isotope dilution ; nitrogen fixation ; nitrogen (N) use efficiency ; nodules ; phosphorus ; phosphorus (P) use efficiency ; roots ; supernodulator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The effect of different levels of soil phosphorus on nodulation, N2 fixation and growth of supernodulating soybean mutant, nts 382, its parent, Bragg, and soybean cultivar, Chippewa, were compared in a greenhouse study. The P rates were, 0, 30, 60 and 90 mg kg−1 soil (P0, P1, P2 and P3, respectively). The reduced shoot growth of nts 382 (25% at P0 and 17% at P3) could not be attributed to P deficiency, as nts 382 achieved near maximum yield at a lower P rate (P1) than Bragg (P2). The P response of Chippewa that produced the lowest shoot yield, was similar to that of Bragg. Root growth of nts 382 was more retarded than shoot growth (on average 50% that of Bragg), and yet absorbed a similar amount of total P as Bragg. Also, Chippewa that had the lowest root dry matter accumulated the highest P in the shoots. The data indicate that plant growth was influenced by differences in P use efficiency rather than differences in total P. In contrast to plant growth, nts 382 accumulated 26% more N than Bragg, and Chippewa accumulated a similar amount of N as Bragg. Thus again, Bragg used N most efficiently for growth. Soil N uptake was not governed by root biomass, and Chippewa absorbed more N from soil than both Bragg and nts 382 which absorbed similar amounts of soil N. Nodulation in nts 382 was significantly enhanced, being 528 and 892% greater than Bragg at P0 and P3, respectively, and nts 382 was the only cultivar whose nodulation was consistently enhanced by P addition. The percentage of N derived from fixation (% Ndfa) was not affected by supernodulation, but because of a greater accumulation of total N in nts 382, amount of N fixed (Ndfa) was on average 30% higher in nts 382 than Bragg. Also, while % Ndfa in either nts 382 or Bragg was not influenced by P addition, total N and consequently, Ndfa increased. However, for Chippewa which fixed the lowest N, both % Ndfa and Ndfa were increased by P addition. We conclude that although supernodulation had an adverse effect on shoot and root growth, these reductions could not be attributed to a reduced uptake of the two major plant nutrients, N and P, neither was there evidence that the mutation had caused a greater demand for these nutrients, other than for the greater requirement of P for nodulation. ei]{gnR O D}{fnDixon}
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  • 82
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    Keywords: Azospirillum ; inoculation ; nitrogen fixation ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Azosprilla were collected in wheat fields from subtropical and temperate soils of central Nepal at various elevations. Different wheat cultivars responded positively and significantly in grain yield, grain N-yield, and total N-yield in plant shoots to the inoculation with Nepalese isolate Azospirillum 10SW. Nepalese wheat cv. Seto responded significantly better with Azospirillum 10SW than with the Brasilian isolate A. lipoferum Sp 108 st, a strain which was found highly efficient in earlier experiments with German wheat cultivars, especially cv. Turbo. Yield of Turbo was increased by inoculations of both Azospirillum strains too, but it showed no significant differences depending from the inoculum used. The higher efficacy of combining Azospirillum 10SW and Seto, both collected from the same locality, indicates the possibility of improved associations using traditional cultivars and local bacteria. ei]{gnR O D}{fnDixon}
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  • 83
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    Plant and soil 151 (1993), S. 147-150 
    ISSN: 1573-5036
    Keywords: Acacia ; isotope dilution ; nitrogen fixation ; N15 ; nitrogen partitioning ; tree
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract An experiment was conducted in the field to determine the partitioning of 15N from ammonium sulfate fertilizer in two-year old trees of Acacia mangium and Acacia auriculiformis. For both species, most of the 15N excess was partitioned into woody tissue, green material and root biomass present at the beginning of the experiment. Approximately one third of the 15N excess was contained in leaves that developed during the 60 days of the experiment. The concentration of 15N excess in the newly developed leaves indicated that more than 90% of the N came from a source other than the fertilizer. Based on the large quantity of N applied in the fertilizer and the inherently infertile soil, it appears that much of the N contained in the newly developed leaves came from N remobilized from other tree parts. ei]{gnR O D}{fnDixon}
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  • 84
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    Plant and soil 155-156 (1993), S. 329-332 
    ISSN: 1573-5036
    Keywords: 15N2 actinorhizals ; controlled environment ; legumes ; nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A gas-tight chamber has been constructed to calibrate the 15N isotope dilution method against direct 15N2 measurements. The theoretical basis for such estimates is given, and the practical problems associated with the experiments are discussed.
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  • 85
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    Plant and soil 154 (1993), S. 211-217 
    ISSN: 1573-5036
    Keywords: nitrogen fixation ; plant growth promotion ; Pseudomonas sp. ; Rhizobium leguminosarum ; vitamins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Fluorescent Pseudomonas sp. strain 267 promotes growth of nodulated clover plants under gnotobiotic conditions. In the growth conditions (60 μM FeCl3), the production of siderophores of the pseudobactin-pyoverdin group was repressed. Plant growth enhancement results from secretion of B vitamins by Pseudomonas sp. strain 267. This was proven by stimulation of clover growth by naturally auxotrophic strains of Rhizobium leguminosarum bv. trifolii and marker strains E. coli thi- and R. meliloti pan- in the presence of the supernatant of Pseudomonas sp. strain 267. The addition of vitamins to the plant medium increased symbiotic nitrogen fixation by the clover plants.
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  • 86
    ISSN: 1573-5036
    Keywords: intercropping ; nitrogen fixation ; nodulation ; pigeonpea ; root respiration ; sorghum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A medium-duration pigeonpea cultivar (ICP 1–6) and a hybrid sorghum (CSH 5) were grown on a shallow Alfisol in monocropping and intercropping systems. Using a monolith method, spatial distribution of nodulation, acetylene reduction activity (ARA) and root respiration were measured. The number, mass and ARA of nodules decreased exponentially with distance from the plant base except at the late reproductive stage. Nodulation and ARA tended to be higher in the intercrop than in the monocrop. Respiration rate of roots increased with distance from the plant base and reached a maximum value at about 20–30 cm. The rate was higher in pigeonpea than in sorghum and also higher in intercrop than in monocrop. This study suggests that pigeonpea roots are physiologically more active than sorghum roots, implying that pigeonpea may become a strong competitor for nutrients in the soil when intercropped. The nitrogen-fixing ability of pigeonpea may be enhanced by intercropping because the sorghum rapidly absorbed inorganic N which would otherwise inhibit N2 fixation.
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  • 87
    ISSN: 1573-5036
    Keywords: Bradyrhizobium japonicum ; Glycine max ; 15N isotope dilution ; nitrogen fixation ; nodulation ; short season ; soil temperature ; soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In areas with a short growing season the poor adaptability of soybean [Glycine max Meer. (L.)] to cool soil conditions is considered the primary yield limiting factor. Soybean requires temperatures in the 25 to 30°C range for optimum N2-fixation and yield. Field studies were conducted in 1990 and 1991 at Montreal, Quebec to determine whether adaptability to cool soil conditions, with respect to earlier symbiosis establishment and function, existed among either Bradyrhizobium strains or soybean genotypes. An early maturing isoline of the soybean cultivar Evans and the cultivar Maple Arrow were inoculated with one of four strains isolated from the cold soils of Hakkaido, northern Japan, or the commercially used strains 532C or USDA110, at two planting dates. Plot biomass and nodulation were assessed at seedling (V2), and flowering(R2) growth stages and harvest maturity. Soybean genotypes did not differ for pre-flowering nodulation or N2-fixation in the cool spring conditions of the first year. Seasonal N2-fixation rates were also determined at the final harvest by the N-balance and 15N-isotope dilution methods. Significantly higher symbiotic activity was found for two of the four Hakkaido strains and was reflected in higher final soybean seed yield and total N2-fixation for the growing season, as compared to the two commercial strains. Planting 14 days earlier resulted in greater early vegetative and total seasonal N2 fixation and yield in the second year when soil temperatures were warmer, emphasizing the need for the development of soybean-Bradyrhizobium combinations superior in nodule development and function under cool soil conditions.
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  • 88
    ISSN: 1573-5036
    Keywords: Acacia mangium ; acidity ; aluminium ; Bradyrhizobium ; Faidherbia albida ; nitrogen fixation ; selection ; symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract This work was designed to determine the role of the acidity and aluminium stress in the selection of partners in the Acacia symbioses with relevance to the persistence of the microsymbiont Bradyrhizobium in the soil and the growth and nodulation of the host plant respectively. Fifteen strains of Bradyrhizobium from Acacia mangium and Faidherbia albida formed a very homogenous acid tolerant group as indicated by their ability to grow better in a medium at pH 4.5 than in a medium at pH 6.8. By contrast, a growth experiment using an acid liquid media (pH 4.5), containing different concentrations of aluminium successfully identified strains sensitive to aluminium toxicity and those able to grow even in the presence of 100 μM AlCl3. Our results suggest that high amounts of aluminium in the soil rather than acidity (pH 4.5) were a major soil factor for selection of Bradyrhizobium strains capable of establishing a permanently high population under natural conditions. Unlike the behaviour of the microsymbiont, growth and nodulation of Acacia mangium and Faidherbia albida were not affected by aluminium, even at 100 μM, but they might be significantly affected by medium acidity (pH 4.5) depending on plant provenances. It is therefore suggested that ability of the host plant to tolerate acidity stress should be taken into account first when screening effective Acacia-Bradyrhizobium combinations for use in afforestation trials.
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  • 89
    ISSN: 1573-5036
    Keywords: genetics ; ineffective nodulation ; mutants ; nitrogen fixation ; symbiosis ; Rj4 gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The dominant allele Rj4 in soybean interdicts or restricts the nodulation of plants by certain strains of bacteria, most of which are classified as Bradyrhizobium elkanii, while the recessive allele permits normal nodulation with the same strains. The near isogenic lines BARC-2 (Rj4) and BARC-3 (rj4) are calculated to be 99.95% identical in their nuclear DNA, but differ specifically in the allele present at the Rj4 locus. These lines were used to identify spontaneous mutants of the Rj4-restricted Bradyrhizobium elkanii strain USDA 61 Nalr that had the ability to effectively nodulate plants of the Rj4 genotype. Of the eight rare nodules found on roots of soybean plants of the Rj4 genotype inoculated with the genetically marked strain USDA 61 Nalr, four were identified as containing mutants with the ability to overcome the effects of the Rj4 allele.
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  • 90
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    Plant and soil 152 (1993), S. 1-17 
    ISSN: 1573-5036
    Keywords: grain legumes ; N2 ; 15N isotope dilution ; nitrogen fixation ; Rhizobium ; symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Biological nitrogen fixation of leguminous crops is becoming increasingly important in attempts to develop sustainable agricultural production. However, these crops are quite variable in their effectiveness in fixing nitrogen. By the use of the 15N isotope dilution method some species have been found to fix large proportions of their nitrogen, while others like common bean have been considered rather inefficient. Methods for increasing N2 fixation are therefore of great importance in any legume work. Attempts to enhance nitrogen fixation of grain legumes has been mainly the domain of microbiologists who have selected rhizobial strains with superior effectiveness or competitive ability. Few projects have focused on the plant symbiont with the objective of improving N2 fixation as done in the FAO/IAEA Co-ordinated Research Programme which is being reported in this volume. The objective of the present paper is to discuss some possibilities available for scientists interested in enhancing symbiotic nitrogen fixation in grain legumes. Examples will be presented on work performed using agronomic methods, as well as work on the plant and microbial symbionts. There are several methods available to scientists working on enhancement of N2 fixation. No one approach is better than the others; rather work on the legume/Rhizobium symbiosis combining experience from various disciplines in inter-disciplinary research programmes should be pursued.
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    Plant and soil 152 (1993), S. 81-85 
    ISSN: 1573-5036
    Keywords: grain legumes ; mutagenesis ; nitrogen fixation ; plant breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Genetic variation among existing cultivars and in germplasm collections is the outcome of selection during evolution and plant breeding. Mutagenesis offers the plant breeder a chance to tackle unconventional objectives, particularly those that were at a selection disadvantage in the past. Effective mutagens are available, but the bottleneck is the effective selection of rare desired variants from large mutagenized populations. Selection methods must be non-destructive. Grain legume mutation breeding has already led to improved cultivars with higher yield, better grain quality, or stronger resistance to pathogenens. Many mutations affecting nitrogen fixation related traits have also been reported. Some could be useful in breeding better cultivars, but the majority are being used to study the factors interacting in the complex process of symbiotic nitrogen fixation and to improve the strategy for producing cultivars with better fixation capacity.
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  • 92
    Electronic Resource
    Electronic Resource
    Springer
    Plant and soil 152 (1993), S. 103-106 
    ISSN: 1573-5036
    Keywords: acetylene reduction ; 15N isotope ; nitrogen fixation ; nodulation ; Phaseolus vulgaris L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Nitrogen fixation in nine common bean (Phaseolus vulgaris L.) lines was estimated using the 15N isotope dilution method at two locations in two seasons. In the first season at one location no N2 fixation was detected while in the second season up to 51 kg N ha−1 were estimated. There were significant differences between lines and correlations between trials were significant for the amounts of N2 fixed, but not for total shoot nitrogen. The plants that fixed the most nitrogen nodulated rapidly after germination. Differences in maximum nodule mass, but not specific nodule activity, were detected also.
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  • 93
    ISSN: 1573-5036
    Keywords: bean ; 15N isotope ; nitrogen fixation ; N fertilizer ; phosphorus ; potassium ; Rhizobium tropici ; sulphur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Although common bean (Phaseolus vulgaris L.) has good potential for N2 fixation, some additional N provided through fertilizer usually is required for a maximum yield. In this study the suppressive effect of N on nodulation and N2 fixation was evaluated in an unfertile soil under greenhouse conditions with different levels of soil fertility (low=no P, K and S additions; medium = 50, 63 and 10 mg kg−1 soil and high = 200, 256 and 40 mg kg−1 soil, respectively) and combined with 5, 15, 60 and 120 mg N kg−1 soil of 15N-labelled urea. The overall average nodule number and weight increased under high fertility levels. At low N applications, nitrogen had a synergistic effect on N2 fixation, by stimulating nodule formation, nitrogenase activity and plant growth. At high fertility and at the highest N rate (120 mg kg−1 soil), the stimulatory effect of N fertilizer on N2 fixation was still observed, increasing the amounts of N2 fixed from 88 up to 375 mg N plant−1. These results indicate that a suitable balance of soil nutrients is essential to obtain high N2 fixation rates and yield in common beans.
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  • 94
    ISSN: 1573-5036
    Keywords: Bradyrhizobium japonicum ; Glycine max ; nitrogen fixation ; nodulation ; rhizobitoxine ; soil nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Foliar chlorosis of soybean (Glycine max [L.] Merr.) resulting from nodulation by rhizobitoxine-producing (RT+) strains of Bradyrhizobium japonicum is commonly less severe in the field than under greenhouse conditions. Differences in nutritional conditions between the field and greenhouse may contribute to this phenomenon. In particular, field-grown plants obtain some N from soil sources, whereas in the greenhouse soybean is often grown in low-N rooting media to emphasize symbiotic responses. Therefore, we examined the effect of NO3 - on the expression of RT-induced symptoms. Soybean plants inoculated with RT+ bradyrhizobia were grown for 42 days in horticultural vermiculite receiving nutrient solution amended with 0.0, 2.5, or 7.5 mM KNO3. Foliar chlorosis decreased with increasing NO3 - application whereas nodule mass per plant was generally increased by NO3 - application. Total amounts of nodular RT remained constant or increased with NO3 - application, but nodular concentrations of RT decreased. Chlorosis severity was negatively correlated with shoot dry weight, chlorophyll concentration, and total shoot N content. It was concluded that application of NO3 - can reduce the negative effects of RT production on the host plant. This suggests that any NO3 - present in field soils may serve to limit chlorosis development in soybeans.
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  • 95
    ISSN: 1573-5036
    Keywords: common bean ; N2 ; 15N isotope dilution ; nitrogen fixation ; Phaseolus vulgaris L. ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Field experiments were performed in Austria, Brazil, Chile, Colombia, Guatemala, Mexico and Peru as part of an FAO/IAEA Co-ordinated Research Programme to investigate the nitrogen fixing potential of cultivars and breeding lines of common bean (Phaseolus vulgaris L.). Each experiment included approximately 20 bean genotypes which were compared using the 15N isotope dilution method. Great differences in nitrogen fixation were observed between and within experiments, with average values of 35% N derived from atmosphere (% Ndfa) and highest values of 70% Ndfa being observed. These values which were larger than had been reported previously for common bean, were observed only when environmental factors were favorable. Therefore, common bean lines are available, which can support high biological nitrogen fixation. These can be used either directly as cultivars for production or in breeding programmes to enhance nitrogen fixation in other cultivars.
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  • 96
    ISSN: 1573-5036
    Keywords: breeding ; nitrogen fixation ; nodulation ; 15N isotope ; Phaseolus vulgaris L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract N2 fixation in lines of Phaseolus vulgaris was measured by 15N-isotope dilution to determine whether a programme of crossing and recurrent selection had resulted in enhanced nitrogen fixation. In field experiments on an isohyperthermic Aquic Hapludoll soil the amount of N2 fixed by the different lines ranged from 18 to 36 kg ha−1 (32 to 47% of plant N) in 56 days. The quantity of N2 fixed and the proportion of plant N derived from fixation was not significantly greater in the lines selected for N2 fixation (RIZ lines) than parental lines. Total shoot N ranged from 53 to 77 kg ha−1 and partitioning of N to pods differed from 28 to 52% among the lines which all had similar growth habit and duration. Nodulation patterns were also distinct. Nodules formed early (10 to 15 plant−1 at 13 days) in many lines, and smallest amounts of fixation were observed in those lines which nodulated slowly and did not form substantial nodule mass until after 40 days. The screening criteria used in the selection of the RIZ lines had been largely indirect with other factors such as disease resistance also being included. Progress for increasing N2 fixation over good-fixing parental lines such as BAT76 was not significant and it is recommended that more attention be paid to early nodulation, to the use of soils with lower available N and to inter-crossing of lines having different good N2 fixation traits in order to further enhance the potential for N2 fixation in beans.
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  • 97
    ISSN: 1573-5036
    Keywords: A-value method ; legumes ; 15N isotope dilution ; nitrogen fixation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The 15N methods are potentially accurate for measuring N2 fixation in plants. The only problem with those methods is, how to ensure that the 15N/14N ratio in the plant accurately reflects the integrated 15N/14N ratio (R) in soil which is variable in time and with soil depth. However, the consequences of using an inappropriate reference plant vary with the level of N2 fixation and the conditions under which the study was made. For example, the errors introduced into the values of N2 fixation are higher at low levels of fixation, and decrease with increasing rates of fixation. At very high N2 fixation rates, the errors are often insignificant. Also, the magnitude of error is proportional to the rate of decline of the 15N/14N ratio with time. Since N2 fixation in most plants would be expected to below 60%, the question of how to select a good reference plant is still pertinent. In this paper, we have discussed some of the criteria to adopt in selecting reference plants, e.g. how to ensure that the reference plant is not fixing N2, is absorbing most of its N from the same zone as the fixing plant, and in the same pattern with time, etc. In addition, we have discussed 15N labelling materials and methods that are likely to minimize any errors even when the fixing and reference plants don't match well in certain important criteria. The use of slow release 15N fertilizer or 15N labelled plant materials results in slow changes in the 15N/14N ratio of soil, and is strongly recommended. Where 15N inorganic fertilizers are used, the application of the fertilizer in small splits at various intervals is recommended over a one-time application. The problem with the reference crop, which has sometimes discouraged potential users of the 15N methods, is surmountable, as discussed in this paper.
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  • 98
    ISSN: 1573-5036
    Keywords: common bean ; maize ; intercropping ; nitrogen fixation ; 15N isotope ; 15N natural variation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Thirty one selected bean lines were evaluated in the field for ability to support N2 fixation when intercropped with maize which received 0, 30 and 60 kg N ha−1 as ammonium sulphate. The amount of fixed N2 was estimated using the natural variation of 15N and wheat as the standard non-fixing crop. Nitrogen as low as 15 kg N ha−1 at sowing suppressed nodule weight and activity (acetylene reduction activity) but not nodule number, suggesting that the main effect of mineral N was on nodule development and function. 15N data revealed a high potential of the bean genotypes to fix N2, with the most promising ones averaging between 50–60% of seed N coming from fixation. Bean lines CNF-480, Puebla-152, Mexico-309, Negro Argel, CNF-178, Venezuela-350 and WBR22-3, WBR22-50 and WBR22-55 were ranked as good fixers.
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  • 99
    ISSN: 1573-5036
    Keywords: common bean ; foliar N ; nitrogen fixation ; N fertilizer ; 15N isotope ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Common bean (Phaseolus vulgaris L.) is able to fix 20–60 kg N ha−1 under tropical environments in Brazil, but these amounts are inadequate to meet the N requirement for economically attractive seed yields. When the plant is supplemented with N fertilizer, N2 fixation by Rhizobium can be suppressed even at low rates of N. Using the 15N enriched method, two field experiments were conducted to compare the effect of foliar and soil applications of N-urea on N2 fixation traits and seed yield. All treatments received a similar fertilization including 10 kg N ha−1 at sowing. Increasing rates of N (10, 30 and 50 kg N ha−1) were applied for both methods. Foliar application significantly enhanced nodulation, N2 fixation (acetylene reduction activity) and yield at low N level (10 kg N ha−1). Foliar nitrogen was less suppressive to nodulation, even at higher N levels, than soil N treatments. In the site where established Rhizobium was in low numbers, inoculation contributed substantially to increased N2 fixation traits and yield. Both foliar and soil methods inhibited nodulation at high N rates and did not significantly increase bean yield, when comparing low (10 kg N ha−1) and high (50 kg N ha−1) rates applied after emergence. In both experiments, up to 30 kg N ha−1 of biologically fixed N2 were obtained when low rates of N were applied onto the leaves.
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  • 100
    ISSN: 1573-5036
    Keywords: acetylene reduction ; Azospirillum ; nitrogen fixation ; para-nodulation ; plant growth regulators ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Treatment of wheat seedlings with the synthetic auxin, 2,4-dichlorophenoxyacetic acid (2,4-d), induced nodule-like structures or tumours (termed para-nodules) where lateral roots would normally emerge. The formation of these structures promoted increased rates of acetylene reduction at reduced oxygen pressure (0.02–0.04 atm) in seedling inoculated with Azospirillum brasilense, compared to seedlings inoculated without auxin treatment. Fluorescent microscopy, laser scanning confocal microscopy and direct bacterial counts all showed that the 2,4-d treatment stimulated internal colonization of the root system with azospirilla, particularly in the basal region of the nodular structures. Both colonization with azospirilla and acetylene-reducing activity were further stimulated by simultaneous treatment with another synthetic auxin, naphthaleneacetic acid (NAA) and, less reliably, with indoleacetic acid (IAA) and indolebutyric acid (IBA). These auxins produced shortening of many initiated lateral roots, although 20 times the concentration of NAA was required to achieve rounded structures similar to those obtained with 2,4-d. Treatment with NAA, IAA or IBA alone also stimulated colonization with azospirilla and acetylene reduction rates at 0.02 atm oxygen, but less effectively than by treatment with 2,4-d. Such exogenous treatments of wheat seedlings with synthetic growth regulators provide an effective laboratory model for studies on the development of a N2-fixing system in cereals.
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