Influence of sulphur supply on glucose and ATP concentrations of inoculated broad beans (Vicia faba minor L.)

In the present study, the influence of S supply on S concentrations, N₂ fixation, available amounts of glucose and adenylates of Vicia faba minor L. were weekly investigated, starting 6 weeks after sowing. Glucose was determined photometrically in shoots, roots and nodules and in ATP, ADP and AMP by bioluminescence in roots, mitochondria and bacteroids. Sulphur deficiency resulted in significantly lower S concentrations of shoots, roots and nodules, in a reduced N₂ fixation as well as in significantly lower amounts of glucose in shoots and nodules. In roots and bacteroids, S deficiency resulted in lower ATP concentrations, while the influence of S supply on ADP and AMP was less pronounced. With optimum S supply, the available amounts of glucose and ATP were strongly influenced by flower formation and seed development. Dedicated to Prof. Dr. Dr. h.c. W. Werner on the occasion of his 75th birthday.     

Carbon costs of nitrate reduction in broad bean (Vicia faba L.) and pea (Pisum sativum L.) plants

The present study aimed at the assessment of carbon (C) costs for nitrate reduction by measuring the additional CO₂ amounts released from roots of nitrate‐fed plants in comparison with urea‐fed plants. Only roots were suitable for these determinations, because nitrate reduction in illuminated shoots is fed nearly exclusively by reducing equivalents coming directly from photosynthetic processes. Therefore, in a first experiment, the sites of nitrate reduction were determined in nodule‐free broad bean (Vicia faba L.) and pea (Pisum sativum L.) plants grown in pots filled with quartz sand and supplied with KNO₃. The extent of nitrate reduction in the various plant organs was determined by measuring in vitro nitrate reductase activity and in situ ¹⁵NO reduction. Only between 9% and 16% of nitrate were reduced in roots of German pea cultivars, whilst 52% to 65% were reduced in broad bean roots. Therefore, C costs of the process could be determined only in broad bean, using an additional pot experiment. The C costs amounted to about 4.76 mol C (mol N)^–1 or 4 mg C (mg N)^–1, similar to those measured earlier for N₂ fixation. The high proportion of nitrate reduction in shoots of pea plants implies that only very little C is required for this nitrate fraction. This can explain the better root growth of nitrate‐nourished pea plants in comparison with N₂‐fixing organisms, which need C compounds for N₂ reduction in roots. Moreover, a different availability of photosynthates in roots of plant genotypes could explain physiologically the occurrence of “shoot and root reducers” in nature.     

Nodulation and N2 fixation of faba bean (Vicia faba L.) after soil amendment with crop residues

The effect of prior soil amendment with different N sources at 50 mg N (kg soil)^—1 on nodulation and N₂ fixation of faba bean (Vicia faba L. cv. Troy) using wheat (Triticum aestivum L. cv. Star) as reference crop was assessed in a pot experiment. Four treatments viz legume manure (LEGM) as clover shoots, cereal manure (CEREM) as barley straw, N fertilizer (FERT‐N) as Ca(NO₃)₂, and no‐manure control (NOMAN) were investigated consecutively at 45, 70, and 90 days after sowing (DAS). Faba bean nodulated profusely, with an increase on average from 629 nodules per pot at 45 DAS to nearly 2.3‐ and 3.3‐fold at 70 and 90 DAS, respectively. Low nodule numbers and nodule dry matter occurred under FERT‐N and CEREM, whereas high values were found for NOMAN and LEGM. Soil amendment affected percent N₂ fixation in relation to N source and plant age. Highest percent N₂ fixation (≥ 90 %) was found under the lowest N‐supplying amendments, no‐manure, and cereal manure, respectively. FERT‐N depressed N₂ fixation particularly at 45 DAS when N₂ fixation was reduced to as low as 23 %. The rise in N₂ fixation thereafter suggests that faba bean adjusted after depletion of mineral N in the soil. N₂ fixation was also decreased after cereal straw application, even though N concentration in faba bean plants was high. The results indicate that plant residues, both with high and low N concentration, applied to soil to raise its fertility may interfere with N₂ fixation of faba bean.     

Modelling nitrogen fixation of pea ( Pisum sativum L.)

Abstract

Nitrogen fixation was simulated for a leafless variety (Delta) of pea (Pisum sativum L.) in central Sweden. It is assumed that N₂ fixation is basically proportional to root biomass, but limited by high root N or low substrate carbon concentrations. Input data on root carbon and nitrogen were estimated from observations of above-ground biomass and nitrogen. The simulated N₂ fixation was compared with estimated values from observations using the ¹⁵N labelling technique. Test data were taken from pea monocultures and pea-oat mixtures with varying pea biomass levels during 1999. Simulated within-season accumulated N₂ fixation correlated to the estimated N₂ fixation with a correlation coefficient (R ²) of 0.74. For seasonal simulations, the predictability was higher (R ²=0.93). Two alternative non-dynamic models, estimating seasonal N₂ fixation as proportional to above-ground biomass and above-ground N, respectively, gave lower predictability (R ²=0.83 and 0.80, respectively). The models were also applied to a second year (1998) and two other sites by comparison with accumulated N₂ fixation estimated by the Difference method. A halved specific N₂ fixation rate (expressed per unit of root biomass) in 1999, compared with 1998, corresponded to essentially dryer and warmer soil conditions during 1999. It was indicated that the variations in soil moisture were more important than soil temperature. It was concluded that the abiotic responses might be of great importance for modelling N₂ fixation rate under different soil conditions.     

Interaction Effects of Nitrogen and Phosphorus on Nodulation in Red Clover (Trifolium pratense L.)

Red clover (Trifolium pratense L.) is one of the most important plants in forage production, especially in northern areas. Fertilisation practices are focused on high yield and forage quality but effects of nutrients on nodulation and N₂ fixation are poorly understood. The aim of this work was to study how nitrogen (N) and phosphorus (P) separately as well as in combination affected nodulation. Red clover plants were grown in pots with gravel in a greenhouse for 11 weeks. To resemble field conditions the root temperature was kept lower than the shoot temperature. Plants were given five different combinations of N and P concentrations during growth. The result showed that at high N concentrations P had a counteracting effect on the N inhibition. The N₂-fixation parameters, nodule number, nodule dry matter and specific nitrogenase activity, were six times higher in plants grown with high N and high P than in plants with high N and low P. When the N₂-fixation parameters and the dry matter of roots and shoots were related to total plant dry matter, there was a stronger effect of P on nodulation parameters than on roots and shoots. This indicates that P has a direct effect on the N₂-fixation parameters, rather than an indirect effect via increased plant growth. These results demonstrate the importance to studying the effects of more than one nutrient at a time.     

Nitrogen fixation and CO2 exchange in soybeans (Glycine max L.) inoculated with mixed cultures of different microorganisms

N₂ fixation, photosynthesis of whole plants and yield increases in soybeans inoculated with mixed cultures of Bradyrhizobium japonicum 110 and Pseudomonas fluorescens 20 or P. fluorescens 21 as well as Glomus mosseae were found in pot experiments in gray forest soil carried out in a growth chamber. The effects of pseudomonads and vesicular-arbuscular (VA) mycorrhizal fungus on these parameters were found to be the same. Dual inoculation of soybeans with mixed cultures of microorganisms stimulated nodulation, nitrogenase activity of nodules and enhanced the amount of biological nitrogen in plants as determined by the ¹⁵N dilution method in comparison to soybeans inoculated with nodule bacteria alone. An increased leaf area in dually infected soybeans was estimated to be the major factor increasing photosynthesis. P. fluorescens and G. mosseae stimulated plant growth, photosynthesis and nodulation probably due to the production of plant growth-promoting substances. Increasing phosphorus fertilizer rates within the range of 5–40 mg P 100 g^-1 1:1 (v/v) soil: sand in a greenhouse experiment led to a subsequent improvement in nodulation, and an enhancement of N₂ fixation and yield in soybeans dually inoculated with B. japonicum 110 and P. fluorescens 21. These indexes were considerably higher in P-treated plants inoculated with mixed bacterial culture than in plants inoculated with nodule bacteria alone.     

Nitrogen fixation of Sesbania rostrata: Contribution of stem nodules to nitrogen acquisition

Abstract

Nitrogen contents, nodule numbers, and nodule dry weights of 6-week-oId Sesbania rostrata plants grown in sand culture with only root nodules, only stem nodules or with both were compared and the root nodules were found to contribute to nitrogen acquisition more significantly than the stem nodules. Similar findings were obtained in ¹⁵N₂-fixing experiments. An 8-week-old plant with both stem and root nodules fixed 1.50 mg nitrogen in a 12 h light period, while the fixation decreased to 1.15 mg nitrogen after the removal of the stem nodules, suggesting that root nodules played major role in nitrogen fixation. However, acetylene-reducing activities per nodule dry weight were higher in the stem nodules. Under flooding conditions, the aerenchyma tissues contributed to about 40% of N₂ transport to root nodules, and 60% was supplied through stem.     

Nitrogenase divarication with histological attributes distinctive for glycosylated aerial stem nodules and nitrosylated root nodules of sesbania

Exceptional symbiotic nitrogen fixation with Sesbania has provided high soil fertility for many past centuries of paddy rice production. Unique stem nodulation results in high nitrogenase activity levels of S. rostrata, Brem, during rapid growth in continuously flooded rice fields that greatly disfavor legume root nodulation and this functional development. The objective of this study was to determine plant nutrient interactions that influence contrasting root and aerial stem nodule histology governing effective nitrogenase activity levels and nitrogen fixation. Top growth, nodulation, and nitrogenase activity levels were significantly increased with increased available soil P. Response to K levels and Ca additions resulted only when soil P was adequate in all treatment combinations. However, there was no significant correlation between fresh nodule weight, nitrogenase activity, and nodules plant^‐1 for both root nodules and aerial stem nodules. Nodule histology was highly contrastive with nodule type and Rhizobium morphology, cytosol composition, and governing enzyme activity levels. Distinctive nonpleomorphic cocci bacteroids of functional aerial stem nodules have tentative designation as Azorhizobium caulinodans gen. nov. sp. nov.     

Genotypic Variation in Nodule Iron Content of Common Bean (Phaseolus Vulgaris L.) in Response to Phosphorus Deficiency

Common bean (Phaseolus vulgaris L.) can supply all of the iron that humans require for metabolism. Also, it fixes atmospheric nitrogen (N₂) in symbiosis with rhizobia. In order to analyze the relation between phosphorus (P) and iron (Fe) elements in nodules and their roles for the plant N₂-dependent growth, six common bean recombinant inbred lines (RIL) of the cross of BAT477 and DOR364 were inoculated with Rhizobium tropici CIAT 899 (originating from International Center of Tropical Agriculture, Colombia) and grown with sufficiency versus deficiency P supply in hydroaeroponic culture. Under P deficiency, the Fe content in nodules decreased in all studied genotypes and was significantly the highest for RIL 34. The nodule contents of Fe and P were significantly correlated under P deficiency. It is concluded that the regressions of nodule Fe content as a function of P content in nodules, roots and shoots, depend upon P supply and genotype.     

Der Einfluß gesteigerter Stickstoffgaben auf die Entwicklung der Wurzelknöllchen,auf die symbiontische Stickstoffassimilation sowie auf das Wachstum und den Ertrag von Ackerbohnen (Vicia faba L.)

The effect of increasing nitrogen fertilizer rates on the development of root nodules, on the symbiotic N₂ assimilation, and on growth and yield of broad beans (Vicia faba L.) In pot experiments with broad beans (Vicia faba minor) the effect of increasing N fertilizer rates (0–800 mg N/pot) on nodulation, symbiotic N assimilation, plant growth, and yield has been studied. The plants were harvested at the beginning and at the end of flowering and at maturity. The following results were obtained: 1. No significant yield differences between treatments were found at the 1st and 2nd harvest, with exception of the 800 mg N/pot treatment which gave higher root yields at the first harvest. 2. At the 1st harvest (beginning of flowering) root nodule yield of the N-zero treatment was higher than the root nodule yield of the N treated plants. At later stages, however, no major differences in root nodule yield between the treatments N₁₀₀ and N₂₀₀ and the N-zero treatment were obtained. 3. Highest grain yields were obtained in the treatment with the lowest N-rate (100 mg N/pot) and in the treatment with the highest N rate (800 mg N/pot). The absolute highest amount of symbiotically fixed N was produced in the low N treatment (100 mg N/pot). Provided that the soil is low in available N a low fertilizer rate is required in order to exploit the full N₂ assimilation potential. 4. The total soil N remained fairly constant throughout the growing period.     

Efficiency of N2 fixation in Vicia faba L. in combination with different Rhizobium leguminosarum strains

The efficiency of symbiotic dinitrogen (N₂) fixation in Vicia faba L. in combination with 3 different Rhizobium leguminosarum strains was studied in a pot experiment during vegetative and reproductive growth. The objective of the experiments was to assess variability among Rhizobium strains inoculated on single legume species and determine possible reasons for observed variations. Dry matter formation, N₂ fixation and the carbon (C) costs of N₂ fixation were determined in comparison with nodule free plants grown with urea. Nodule number and the capacity of different respiratory chains in the nodules were also measured. The plants inoculated with the Rhizobium strain A 37 formed less dry matter and fixed less N compared to the other two Rhizobium strains (Vic 1 and A 150). This coincided with a lower number of nodules and higher C costs of N₂ fixation. The C costs for N₂ fixation were in all cases significantly lower during reproductive growth compared to vegetative growth. Neither the latter nor the differences in C expenditure for N₂ fixation between the Rhizobium strains could be explained in terms of differences or shifts in the capacity of different respiratory chains in the nodules.     

Effects of hairy vetch foliage application on nodulation and nitrogen fixation in soybean cultivated in three soil types

We investigated the effects of applying hairy vetch foliage on nodulation and atmospheric nitrogen (N₂) fixation in soybean cultivated in three soil types in pot experiments. Soybean plants were grown in Gley Lowland soil (GLS), Non-allophanic Andosol (NAS), and Sand-dune Regosol (SDR) with hairy vetch foliage application in a greenhouse for 45 days. In GLS, the nodule number was not influenced by the application, however, nodule dry weight and N₂ fixation activity tended to increase. In NAS and SDR, nodule formation was depressed by foliage application. Soybean plant growth was promoted in GLS and SDR but not in NAS. These promotive effects of hairy vetch foliage application on soybean plant growth in GLS were considered to be mainly caused by the increase in N₂ fixation activity of the nodules, whereas it was considered to be mainly caused by the increase in nitrogen uptake activity of the roots in SDR. The varying effects of hairy vetch foliage application on soybean nodulation may be due to soil chemical properties such as pH and cation exchange capacity, which are related to soil texture. Therefore, we conclude that it is important to use hairy vetch for soybean cultivation based on the different effects of hairy vetch on soybean plant growth in different soil types.     

N2 fixation and growth of legumes as affected by sulphur fertilization

The influence of three sulphur application rates in combination with two nitrogen application rates on N₂ fixation and growth of different legumes was investigated. N was applied as N-labelled ¹⁵NH₄ ¹⁵NO₃. The ¹⁵N isotope dilution technique was used to estimate N₂ fixation. At both N increments dry matter yield was highest with high S supply. Independently of the N supply, the high S application rate resulted in a significantly higher N accumulation, which was mainly caused by a higher N₂ fixation rate. With the grain legumes the weight of nodules was increased by the high S application rate. The higher number of nodules per pot with optimum S supply was the result of a better root growth. Rates of acetylene reduction correlated significantly with S supply.     

Inoculation with Phosphate Solubilizing Mesorhizobium Strains Improves the Performance of Chickpea (Cicer aritenium L.) Under Phosphorus Deficiency

The use of phosphate-solubilising bacteria as inoculants increases plant phosphorus (P) uptake and thus crop yield. Strains from the genus Mesorhizobium are among the most powerful phosphate solubilizing microorganisms. In order to study efficiency in P uptake and N₂ fixation in chickpea (Cicer aritenium), forty-two rhizobia strains natively from Tunisian soils were studied in symbiosis with the chickpea variety “Béja1” which is frequently cultivated in Tunisia. Plants were inoculated separately with these strains under controlled conditions in perlite under two sources of P i.e. soluble (KH₂PO₄) and insoluble P (Ca₂HPO₄). At flowering stage, growth, nodulation, P uptake and N₂ fixation were assessed in all symbiotic combinations. The results showed that the S27 strain efficiently mobilized P into plants, observed as a significant increase of plant P content when insoluble P (Ca₂HPO₄) was supplied to the soil. This was associated with a significant increase in plant biomass, nodule number and N content under insoluble P conditions. Additionally, inoculation with the Mesorhizobium strain S27 significantly increased the root acid phosphatase activity under insoluble P. This study also shows significant correlations found between plant P content and acid phosphatase activity under low P conditions which may highlight the contribution of acid phosphatases in increasing P use efficiency. A field experiment also showed that most of the chickpea analyzed parameters were improved when plants inoculated with two selected rhizobia strains (S26 and S27) and supplied with P2O5. Overall, these findings postulate that rhizobial inoculation should not only be based on the effectiveness of strains regarding N fixation, but also to other traits such as P solubilisation potential.     

Plant and soil responses to mycorrhizal fungi and rhizobacteria in nodulated or nitrate-fertilized peas (Pisum sativum L.)

Rhizosphere organisms affect plant development and soil stability. This study was conducted to determine the effects of a vesicular-arbuscular mycorrhizal (VAM) fungus [Glomus mosseae (Nicol. &>; Gerd.) Gerd. and Trappe] and a rhizobacterium (Bacillus sp.) on nitrate-fertilized or nodulated pea (Pisum sativum L.) plants and on the status of water-stable soil aggregates. The plants were grown in pots in a yellow clay-loam soil, and inoculated with the VAM fungus and the rhizobacterium, with one of the two, or with neither. The Bacillus sp. and G. mosseae did not affect shoot dry mass in nodulated plants. Under N fertilization, the VAM fungus enhanced plant growth, while the rhizobacterium inhibited shoot growth, VAM root colonization, and nodule formation, but enhanced the root:shoot and the seed:shoot ratios. The inhibition of shoot growth and of root colonization appeared to be related. The water stability and pH of the VAM soils were higher than those of the non-VAM soils. The rhizobacterium enhanced the water-stable aggregate status in the non-VAM soils only. Under both N-nutrition regimes, the soils had the greatest proportion of the water-stable aggregates when inoculated with both rhizo-organisms and the lowest when colonized by neither. The two rhizo-organisms affected both plants and soil, and these effects were modified by the source of N input through N₂ fixation or fertilization. Received: 5 April 1995     

Nitrogen metabolism is related to improved water‐use efficiency of nodulated alfalfa grown with sewage sludge under drought

Leguminous plants grown in sewage sludge–amended soils can acquire nitrogen by assimilation of nitrate and ammonium from the soil solution or from atmospheric‐dinitrogen (N₂) fixation through association with N₂‐fixing bacteria. We proposed that operation of both metabolic processes could contribute to alleviate the impact of drought in sludge‐treated plants. A greenhouse experiment was conducted to evaluate the involvement of nodule metabolism in the use efficiency of water and N in sludge‐treated plants. Treatments comprised (1) plants inoculated with rhizobia and amended with sewage sludge; (2) plants inoculated with rhizobia without any amendment; and (3) noninoculated plants supplied with ammonium nitrate, each under well‐watered and drought conditions. Under drought, sludge‐treated plants had increased plant growth and higher photosynthetic and water‐use efficiencies than untreated plants. Drought stimulated nitrate reductase and GS/GOGAT activities but did not affect the activities of phosphoenolpyruvate carboxylase and malate dehydrogenase or the leghemoglobin concentration. The results suggest that under drought conditions, both N₂ fixation and nitrate assimilation in nodules of sludge‐treated plants contributed to improve plant N supply and to increase the drought tolerance of alfalfa.     

Bedeutung der arbuskulären Mykorrhiza (AM) für die Schwermetalltoleranz von Luzerne (Medicago Sativa L.) und Hafer (Avena sativa L.) auf einem klärschlammgedüngten Boden

Effect of arbuscular mycorrhizal fungi (AMF) on heavy metal tolerance of alfalfa (Medicago sativa L.) and oat (Avena sativa L.) on a sewage-sludge treated soil In pot experiments with a sewage sludge treated soil, the influence of two arbuscular mycorrhizal fungi (AMF) isolates of Glomus sp. (T6 and D13) on plant growth and on the uptake of heavy metals by alfalfa (Medicago sativa L.) and oat (Avena sativa L.) was investigated. Alfalfa showed an increase of biomass with mycorrhizal infection only to a small extent. In oat AMF inoculation increased the growth of both root and shoot by up to 70% and 55% respectively. Mycorrhization raised the P-content and -uptake in alfalfa, but not in oat, in both roots and shoots. Mycorrhizal alfalfa showed lower Zn-, Cd- and Ni-contents and uptake in roots and shoots. The root length was significantly decreased in mycorrhizal alfalfa plants (up to 38%). The translocation of heavy metals into the shoot of mycorrhizal alfalfa was slightly increased. Mycorrhizal infection of oat led to higher concentrations of Zn, Cd and Ni in the root but to less Zn in the shoot. The translocation of heavy metals to the oat shoot was clearely decreased by mycorrhizal colonisation. This may be based on the ability of fungal tissues to complex heavy metals at the cell walls, thus excluding metals from the shoot. This conclusion is supported by the enhanced root length (up to 78%) of mycorrhizal oat plants in this experiment. The mycorrhizal infection seemed to protect plants against heavy metal pollution in soils. It was obvious that different host plants reacted in different ways.     

Nitrogenase Activity and Nodule Biomass of Cowpea (Vigna unguiculata L. Walp.) Decrease in Cover Crop Mixtures

Legume cover crops are often used to build soil nitrogen (N) fertility and there is increasing interest in cover crop mixtures. The objective of this mechanistic greenhouse study was to determine the effect of cover crop community diversity and soil fertility on nitrogenase activity and nodule biomass of cowpea. Cover crops were grown for 42–53 days, aboveground biomass was harvested, and nitrogenase activity was estimated with the acetylene reduction assay. Roots were then excavated to determine nodule and root biomass. Nitrogenase activity and nodule biomass per plant were greatest in cowpea monoculture and reduced by 71–98 percent in four-species mixtures. Reduced capacity for N₂ fixation was partially driven by lower cowpea biomass in mixtures. The ratio of root nodule / shoot biomass increased by 81–297 percent in low-fertility relative to high-fertility soils, which contributed to increased nitrogenase activity. Results suggest cowpea monocultures in low-fertility soils have the greatest potential for N₂ fixation.     

Iron dependent ureides and allantionase synthesis in pigeon pea

The effect of iron (Fe) on ureide metabolism was examined in 45‐day‐old pigeon pea (Cajanus cajan L.) (ureide plant) and alfalfa (Medicago sativa) (amide plant). Plants were either inoculated with Rhizobium or fertilized with ammonium nitrate (NH₄NO₃). The ureides, allantoin and allantoate, and allanotinase activity were increased in pigeon pea with Fe supplementation. Specific effect of Fe on ureide metabolism of pigeon pea was indicated by the lack of same effect in alfalfa under similar conditions. Nitrogenase activity was elevated with increasing concentrations of Fe in pigeon pea (ureide) as well as alfalfa (amide) symbiosis. Nitrogen (N₂) fixation, ureides, and allantoinase activity were reduced at 10 ppm and above concentration of Fe.