Soil fertility effects on growth,nitrogen fixation,nodule enzyme activity and xylem exudates of lablab purpureus (L.) sweet,grown on a typic eutrustox

Abstract

The Lablab or Hyacinth bean, Lablab purpureus (L.) Sweet, is a drought tolerant grain‐forage legume widely grown within a wide range of neotropical regions of the world. It has been an important cultivated food grain crop for many centuries within extensive warm humid‐subhumid climatic areas and grown on widely different Ultisol and Oxisol soil types. The objective of these studies was to determine soil fertility effects of Ca, P, and K factorial combinations applied to a Typic Eutrustox on growth, nodulation, and nitrogen fixation of Lablab bean. Stem and nodule xylem components were determined along with soil pH and nitrification within the nodule rhizosphere. Highly significant increases resulted with P fertilization for stem, leaf, seed, nodulation, and nitrogenase activity. With the exception of nitrogenase, all were also increased with K additions. Both stem and seed were significantly increased with Ca treatments. Highly significant increases for total plant N content resulted with P, K, Ca, and P x K interaction. Allantoinates were initially dominant components of stem xylem exudate following decapitation but were not detected after 5 days. Amino‐amide composition remained relatively constant. Nitrates increased from 1.6 to 81.8 ug N ml^‐1 exudate during the 8 day collection period. Soil nitrate levels adjunct to decomposing nodules increased from 4.2 to 1661.1 ug g^‐1 soil. Initial high levels of ureide transforming enzymes ALTN, ALTC, and URC were not detected after 4 days. Amine‐amide enzymes AST, GS, GOGAT, and GDH were relatively constant with nitrate reductase increasing from 0.12 to 9.35 IU ml^‐1 xylem exudate during the 8 d period. Nodule xylem export components were dominated by ureides, 429.8 umol, and amines 30.3 umol ml^‐1. Enzyme activity levels were highest for AST 22.17 and GS 13.25 IU ml^‐1 with the ureide enzymes URC 2.24, ALTN 0.26, and ALTC 0.15 IU ml^‐1 . Nodule exudate composition was K 422.0, P 63.4, Ca 53.8, Mg 42.8, and Na 25.6 ug ml^‐1.     

Effects of soil fertility on growth,tuber yield,nodulation and nitrogen fixation of yam bean (Pachyrhizus erosus (L.) urban) grown on a typic eutrustox

The tuberous legume, Yam Bean, (Pachyrhizus erosus (L.) Urban, has been utilized as a food crop for many centuries. The large, starchy tubers have higher aitrogen content than potatoes, cassava and taro roots with 20% or more of the N fraction as ureidoglyco‐lates. Yam Bean (Jicama) tuber growth within the neotropical regions of the world is influenced by soil productivity. The objective of this study was to determine effects of soil fertility treatments on tuber yield, nodulation characteristics and nitrogen fixation.

Highly significant increases in growth and tuber production resulted with P additions, and to Ca and K levels when combined with P. Nodule weight and nitrogenase activity were similar in response to factorial soil treatments as were the tuber and total above ground plant growth. Total ureide content of tuber epiperi‐derm tissue increased significantly with P, Ca, and P + Ca treatments and increased quadratically with increased K additions when combined with P, Ca, and P + Ca treatments. Multiple regression for nitrogenase = 3.4 g top wt. + 3.5 g tuber wt. + 4.9 g nodule wt. + 3.7 umol ureide, R² = 0.88 and C.V. = 16.5%. The percent nonstructural tuber carbohydrates was not significantly influenced by soil treatments although total tuber N content significantly increased with the P treatments. Content of plant nutrient elements within nodule cytosol generally increased significantly with addition of the corresponding element in the soil fertility treatments. Increased K content was quadratic for increased K additions with concomitant decrease in Na content having highly significant negative correlation (r = ‐0.72). Adequate available soil P, Ca and K favorably influenced Yam Bean tuber growth and nitrogen fixation with P a first limiting factor for desirable high tuber yields.     

Acid soil tolerance of leucaena species in greenhouse trials

Abstract

Lines of Leucaena leucocephala (Lam.) de Wit were grown in greenhouse pots of an acid, Al‐toxic Tatum subsoil (clayey, mixed, thermic typic Hapludult) treated with 0 or 3000 ppm CaCO₃ to give final soil pH values of 4.1 and 5.3, respectively. Lines of L. leucocephala, plus those of other Leucaena species, were also tested on an acid, Monmouth soil (clayey, mixed, mesic, typic Hapludult) treated with 0 or 1500 ppm CaCO₃ to give final soil pH values of 4.8 and 6.6, respectively. The major index of acid soil tolerance used was relative root yield (unlimed/limed %).

Relative root yields of 117 L. leucocephala lines on Tatum soil ranged from 34 to 246%. Hence, liming the soil from pH 4.1 to 5.3 was highly beneficial to some lines and highly detrimental to others. Because Tatum subsoil is 89% Al saturated at pH 4.1, line tolerance to unlimed soil indicates tolerance to Al. Causes of yield depression at pH 5.3 were not determined.

On Monmouth soil, in a test involving 148 lines of 6 Leucaena species, relative root yields (unlimed/limed %) ranged from 23 to 386%. The line showing highest tolerance to the acid soil (P.I. 279578) and that showing lowest tolerance (P.I? 281636) are both L,. leucocephala. The majority of lines used on Monmouth soil (124 of a total of 148) were from this species. Average performances of the 6 species indicated that L. diversifolia Benth. (5 lines) was most tolerant to the acid Monmouth soil and liming the soil from pH 4.8 to 6.6 actually decreased root yields. The species L.. leucocephala (124 entries) and L. pulverulenta Benth. (4 lines) were intermediate, and L. lanceolata S. Wats. (3 lines) and I., retusa Benth. (1 line) appeared more sensitive to acid Monmouth soil. The Al saturation of Monmouth soil at pH 4.8 was only 23% (compared with 89% for Tatum at pH 4.1). The major growth limiting factor in acid Monmouth soil is believed to be Al toxicity, but this soil has not been as throughly characterized as has Tatum, and other factors may well be involved in explaining differential tolerances of Leucaena lines on the unlimed versus limed soil.

Results of these studies indicate that Leucaena species and lines within species differ significantly in tolerance to acid soils having high levels of exchangeable Al. Acid soil tolerant lines of Leucaena may be useful in expanding the acreage of this crop on oxisols and ultisols of the tropics and subtropics.     

Soil fertility effects on growth,yield, nodulation and nitrogenase activity of Austrian winter pea

Austrian winter pea (Pisum sativum subspecies arvense (L.) Poir) is grown as a cool season annual to produce high protein seed and forage as well as for soil fertility improvement. This legume is grown on a wide range of soil types with many different cropping systems. The objective of these studies was to determine the influence of K levels, with and without P and Ca fertilization, for increased growth, yield, nodulation and nitrogenase activity. Results were from 3 years’ field and greenhouse experiments with a Psammentic Paleustalf (Eufaula series) utilizing Rhizobium leguminosarum (Frank), ATCC 10314 as inoculum. Soil fertility effects on composition and histology of field‐grown nodules are presented.

Available soil P was a limiting plant nutrient in field studies with significant response to K resulting with PK combinations for top growth, tillers, pods, seed yield, nodule mass, and nitrogenase activity levels (C₂H₂, red.). Multiple regression for nitrogenase (umol C₂H₄ h^‐1) = 1.09 tiller number + 3.37 nodule weight + 2.29 pod number, R² = 0.837, C.V. = 29.9%. Results from the greenhouse experiments indicated significant responses with increased K application levels when combined with P and Ca fertilization for top growth, nodule weight, number of nodules and nitro‐genase activity. Highly significant correlations resulted with nitrogenase x nodule weight (r=0.538) and nitrogenase x top growth (r=0.359) with multiple regression of treatment effects for nitrogenase (μmol C₂H₄ h^‐1) = 2.73 P + 1.04 K + 4.92 Ca, R² = 0.797 and C.V. = 48.8%. Soil addition of plant nutrients resulted in significantly increased concentrations of those elements within nodules. Magnesium content was not consistently influenced by P, Ca, and K amendments. Sodium decreased with increased K fertilization. Multiple regression of elemental composition (mg g^‐1 nodule) for nitrogenase (pmol C₂H₄ h^‐1) = 0.21 P + 0.86 K + 2.35 Ca ‐ 2.01 Na, R² = 0.772, C.V. = 55.6%. The proportion of plant nutrients in nodules contained within the nodule cytosol was highest for K (56.2%) and lowest for Ca (21.4%) with intermediate levels of Mg (50.2%), P (45.4%), and Na (37.2%).

Practical application from these data include the requirement of adequate available soil K for increased yield and nitrogen fixation with favorable P and Ca soil levels in Austrian winter pea production.     

Perennial winged bean yield and nitrogen fixation improvement with soil fertility treatments of a typic eutrustox

The perennial legume, Winged Bean (Psophocarpus tetragonolobus (L) DC), has potential as a high protein food crop for the humid, tropical regions of the world. Edible seed pods, oil seed grain, leaves, flowers, and unique high protein tubers provide abundant nutritious components desirable for improved human diets. However, soil characteristics and fertility levels influence plant growth, yields and nitrogen fixation capability of this legume. Objectives of this study were to determine soil‐plant nutrient influences on vigorous regrowth, pod and tuber yields, nitrogenase (C₂H₂ red.) activity levels, and nodule cytosol components of the perennial Siempre cultivar grown on a Typic Eutrustox during three years, 1978–1980.

Available soil phosphorus was a first limiting plant nutrient during all three years of plant age. Effects of combined 100 mg P with 200 mg K/kg soil were highly significant for every parameter and plant age. Pod and seed yields more than doubled with PK addition compared to the check. Tuber growth, nodule mass and nitrogenase activity levels more than trebled with PK treatments as compared to the check. Both elemental P and K were significantly increased within the nodule cytosol of fertilized plants. Cytosol Na was significantly decreased with soil K additions. The best fit multiple regression was: nitrogenase = 1.99 nodule wt. + 6.34 tuber wt. + 0.39 tuber % N + 5.08 cytosol P + 1.55 cytosol K ‐ 0.45 cytosol Na, R² = 95.5, C.V. = 11.2%. The dominant nodule cytosol enzymes, aspartate aminotransferase (AST) and glutamine synthetase (GS), significantly increased with soil K additions regardless of P treatments. Glutamate dehydrogenase (GDH) and glutamate synthase (GOGAT) also contributed significantly with multiple regression for nitrogenase = 1.07 GS + 2.1 AST + 1.74 GOGAT ‐ 1.76 GDH + 12.89 Ureide, R² = .89, C.V. = 17.3%. Highly significant increases in nodule cytosol ureide composition with K soil additions has interest because of the role as a nitrogenous nonprotein component for many legumes. Increases in growth, nodulation and nitrogenase activity levels resulted with increased K levels of 0, 100, 200 and 300 mg K/kg soil when soil P and Ca were not limiting.     

Soil fertility effects on nodulation,nitrogen fixation,and yields of guar, (Cyamopsis tetragonoloba (L.) Taub.), grown on a typic eutrustox

Guar (Cyamopsis tetragonoloba (L.) Taub.) is a legume of unusual industrial importance due to increased utilization of Guar gum, ‘Guran,’ in numerous modern manufacturing and food processes. Guar has high tolerance of drought and inhibitory soil characteristics and has been utilized for many centuries within the subhumid regions of the Indo‐Pakistan subcontinent. Nodulation, nitrogen fixation, and yields of high protein seed with desirable Guran content are improved by favorable soil fertility. Plant growth, seed yields, and nodulation are influenced by plant population density.

Top Growth quadratically increased with plant population densities of 10, 25, and 50 plants per meter² for P and P+K treatments. Highest seed yield and nodule development resulted for all soil fertility treatments with 25 plant per meter². Ontogenetic response from anthesis to full seed maturity resulted with increased nodulation significantly correlated (r = 0.67) with top growth. However, nitrogenase activity levels plant‘¹ declined significantly with plant maturity. Nodule cytosol enzyme activity levels at anthesis were: allantoinase (ALTN) 29.5 IU, aspartate transaminase (AST) 19.3 IU, glutamine synthase 12.5 IU, uricase (URC) 2.25 IU, and allantoicase (ALTC) 0.14 IU g^‐1 fresh nodule. Cytosol composition included ureidoglycolates (ureides) 425.8 μmol, amine‐amide content 29.5 μmol, nitrate 1.95 μmol, pyruvate 0.21 μimol, and oketoglutarate 0.09 μmol g‘¹ fresh nodule. Plant nutrient composition of nodule cytosol was K 394.9 jig, P 59.4 μg, Ca 48.9 μg, Mg 39. μg, and Na 19.0 μg g^‐1 fresh nodule. Precise histology with Giemsa and PAS procedures were essential for microscopy with cellular compartmentalization of purine enzymatic transformations governing nitrogenase activity and nodule development.     

Relationship between nitrogen fixation and numbers of cyanobionts and heterocysts in fifteen Azolla strains

The relationship between nitrogen fixation and the numbers of cyanobionts and heterocysts in the mature leaves of fifteen Azolla strains from five species was examined. The nitrogenase (acetylene reduction) activity per leaf and the numbers of cyanobionts and heterocysts in the mature region varied depending on the Azolla strains. The numbers of cyanobionts and heterocysts, particularly the number of heterocysts, were closely correlated with the nitrogenase activity per leaf. However, the nitrogenase activity per leaf was not correlated with the nitrogenase activity per heterocyst. The results indicate that the variation in the nitrogenase activity of the leaves among Azolla strains mainly resulted from the variation in the number of heterocysts and not from the variation in the nitrogenase activity per heterocyst.     

Effects of changes in applied nitrogen concentrations on nodulation,nitrogen fixation and nitrogen accumulation during the soybean growth period

ABSTRACT

The specific mechanism by which nitrogen application affects nodulation and nitrogen fixation in legume crops remains uncertain. To further study the effects of nitrogen application on soybean nodulation and nitrogen accumulation, three consecutive tests were performed during the VC-V4, V4-R1 (10 days), and R1-R2 (10 days) growth periods of soybean. In a dual-root soybean system, seedlings on one side were watered with a nutrient solution containing NH₄⁺ or NO₃^? as the N source (N+ side), and those on the other side were watered with a nitrogen-free nutrient solution (N- side). During the VC-R2 period, on the N+ side, high nitrogen treatment inhibited nodule growth and nitrogenase activity (EC 1.18.6.1), and the inhibition was significantly increased with increasing high nitrogen supply time (10 days, 20 days). When the high nitrogen treatment time reached 20 days, the specific nitrogenase activity (C₂H₄ μmol^?1 g^?1 nodule dry mass h^?1) was similar to that in the low nitrogen treatment, indicating that the nitrogen fixation capacity per gram of dry mass nodules was almost the same. Therefore, it is assumed that long-term high nitrogen treatment mainly reduces nitrogen fixation by reducing the nodule number. The effect of nitrogen concentration on the roots on the N+ side was greater than that on the N- side. Taken together, these results indicate that nitrogen application affects a contact-dependent local inhibition of root nodule growth, nitrogenase activity, and nitrogen accumulation. The whole plant systematically regulates specific nitrogenase activity, and high nitrogen inhibition is recoverable.     

Soil fertility effects with tripartite symbiosis for growth,nodulation and nitrogenase activity of vicia faba L

Fababean (Vicia faba L.) is one of the oldest known important grain legume food crops grown within the temperate and subtropical regions of the world. This species is adapted for both forage and food grain production as a cool season annual crop on a wide range of soil and climatic conditions with effective tripartite symbiosis. Both Rhizobium and endophyte mycorrhiza are essential for high levels of production and symbiotic N₂ fixation. The objective of these greenhouse and field studies was to determine effects of Glomus fasciculatum colonization with soil fertility treatments to a Psammentic Paleustalf (Eufaula series) on growth, nodulation, nitrogenase activity and nodule composition for V. faba, var. Major, fuh Rumi (Nile) inoculated with R. leguminosarum Frank.

Top growth and nodule mass were increased approximately 10 fold and nitrogenase activity about 7 fold with the highly significant effect of mycorrhiza and response to low soluble Ca₃(PO₄)₂ fertilization in greenhouse studies. With both effective mycorrhiza and Rhizobium inoculation in the field experiments, seed yields were correlated with top growth (r = 0.841). Phosphorus and Ca fertilization resulted in highly significant increases in seed yields. Nitrogenase activity was correlated with nodule wt. (r = 0.958) and highly significant increases resulted with P and Ca soil amendment. Plant nutrient element composition of nodules increased with the fertilization treatments for P, Ca and increased K levels. Sodium content decreased significantly with increased K fertilization (r = ‐0.846). Potassium composition increased significantly with P content (r = 0.523). Enhanced N₂‐fixation along with increased high protein forage and food grain production with Fababeans have much potential. However, soil fertility and management techniques for improved production include effective mycor‐rhizal colonization.     

Influence of pruning frequency of Albizia lebbeck, Gliricidia sepium and Leucaena leucocephala on nodulation and potential nitrogen fixation

The influence of four pruning frequencies on biomass, nodulation and N₂ fixation was investigated on Albizia lebbeck, Gliricidia sepium and Leucaena leucocephala grown in the screenhouse for 16 months, using acetylene reduction and ¹⁵N dilution methods. Frequent prunings at 4-month intervals had no deleterious effect on symbiotic N₂ fixation, which increased in Gliricidia and Leucaena in particular. Nodulation and nitrogenase activity varied inconsistently within species, and were not influenced by pruning frequency. Cumulative assessment of pruning effect showed higher biomass, N yield and N₂-fixing capacity of the woody species than at last harvest, and appeared to have more practical relevance. Across species, cumulative total dry matter, N yields, and both percentage and absolute amount of N₂ derived from atmosphere increased with pruning frequency, except when trees were pruned 3 times. Of the three species, G. sepium had the lowest biomass production, N₂ fixation and N accumulation. Received: 25 October 1995     

Effects of salt stress on nitrogen fixation,oxygen diffusion,and ion distribution in soybean,common bean,and alfalfa

The exposure of legume nodulated‐roots to 100 mM NaCl resulted in a rapid decrease in plant growth associated with a short‐term inhibition of both nodule growth and nitrogenase activity (C₂H₂ reduction=ARA). However, these NaCl effects varied among species, common bean being more sensitive than soybean and alfalfa. The higher sensitivity of common bean was associated with a higher accumulation of sodium (Na) and chlorine (Cl) in the nodules and only a small difference between salt‐treated and control plants of common bean in their responses of ARA to raising rhizosphere pO₂. By contrast, soybean and alfalfa plants showed a higher stimulation of ARA by pO₂ for the salt‐treatment than for the control. It is concluded that the intraspecific variation in short‐term inhibition of ARA by salt may involve the regulation of O₂ diffusion and the distribution of ions in nodules.     

Effect of Mn on growth,modulation, and nitrogen fixation by soybeans grown in the greenhouse

Abstract

Soybean (Glycine max (L.) Merr. cv Bragg) plants were grown in the greenhouse using a low‐Mn Leefield sand amended with 0, 2.5, 5, 20 and 50 yg Mn/g. The plants were inoculated and were primarily dependent on symbiotically fixed N. Measurements of DTPA‐extractable soil Mn, soil pH, leaf tissue Mn, top weight, top N content, and nodule weight, volume and number were made at 27, 42, 56, 63 and 69 days after planting. The DTPA extrac‐tant was a good predictor of leaf tissue Mn giving a highly significant (P = 0.01) overall correlation coefficient of 0.704 for this comparison. Because of an unexpected decline in soil pH from 6.8 to 6.0 and an associated increase in DTPA‐extractable Mn from 0.14 to 0.24 yg/g during preparation and handling prior to the first harvest time, Mn in the leaf tissue of the controls was never less than 21 yg/g. Since this concentration of Mn is above the deficient level, no significant responses in top growth, nitrogen fixation or nodule measurements were obtained from the addition of low rates of Mn. The highest Mn rate was only mildly toxic in terms of top growth and top N content, producing leaf tissue having Mn concentrations ranging from 171 to 180 yg/g at the last three harvest periods.     

Effect of organic matter on the growth,nodulation, and nitrogen fixation of soybean grown under acid and saline conditions

Abstract

A field experiment was conducted to examine the effects of two organic materials (Bark and Tenporon composts) on the growth, nodulation and nitrogen fixation (measured as acetylene-reducing activity) of soybean plants (Glycine max L.) under acid and saline conditions. These organic materials were incorporated into acid and saline (induced by irrigating the soil with a NaCl solution) soils at 4% rate of application.

These composts tended to improve nodule formation in soybean under acid and saline conditions, especially in the plants treated with Bark compost. Acetylene-reducing activity was significantly (P<0.05) increased by these composts under acid condition.

These composts also tended to improve shoot growth under acid and saline conditions, presumably due to the improvement of the soil moisture status of the soils and nutrient uptake. These results suggest that the growth of soybean could be improved by the application of organic matter under acid and saline conditions.     

Rhizobiophage effects on nodulation,nitrogen fixation,and yield of field-grown soybeans (Glycine max L. Merr.)

Summary Previous laboratory and greenhouse studies have shown that phages significantly reduce soil populations of homologous rhizobia. Reductions in nodulation and N₂ fixation have also been observed. The purpose of the current study was to examine the effect of a phage specific ofBradyrhizobium japonicum USDA 117 on nodulation, nodule occupancy, N₂ fixation and soybean growth and yield under field conditions. The phage was inoculated in combination withB. japonicum USDA 117 and/orB. japonicum USDA 110 (resistant strain) into a rhizobia-free sandy loam soil and planted toGlycine max (L.) Merr. Williams. When the phage was applied to soil inoculated withB. japonicum USDA 117 alone, significant reductions in nodule weight and number, shoot weight, foliar N, nitrogenase activity, and seed index were observed. When, however, the soil also contained the non-homologous strain,B. japonicum USDA 110, no significant effects on any of these parameters were found. Nodule occupancy by competing strains ofB. japonicum USDA 110 and USDA 117 was also affected by the phage. In soil which did not contain the phage, 46% and 44% of the identified nodules were occupied by USDA 110 and 117, respectively. When the phage was present in the soil, nodule occupancy byB. japonicum USDA 117 was reduced to 23%, while occupancy byB. japonicum USDA 110 was increased to 71%. These results suggest that nodulation by selected strains of rhizobia can be restricted and nodulation by more effective, inoculated strains can be increased through the introduction of a homologous phage to soils.     

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.     

Rapid and reversible nitrate inhibition of nodule growth and N2 fixation activity in soybean (Glycine max (L.) Merr.)

Nodulated soybean (Glycine max. (L) Merr. cv. Williams) plants were hydroponically cultured, and various combinations of 1-week culture with 5 or 0 mm nitrate were applied using 13-d-old soybean seedlings during three successive weeks. The treatments were designated as 0-0-0, 5-5-5, 5-5-0, 5-0-0, 5-0-5, 0-5-5, and 0-0-5, where the three sequential numbers denote the nitrate concentration (mm) applied in the first-second-third weeks. The size of the individual nodule was measured periodically using a slide caliper. All the plants were harvested after measurement of the acetylene reduction activity (ARA) at the end of the treatments. In the 0-0-0 treatment, the nodules grew continuously during the treatment period. Individual nodule growth was immediately suppressed after 5 mm nitrate supply. However, the nodule growth rapidly recovered by changing the 5 mm nitrate solution to a 0 mm nitrate solution in the 5-0-0 and 5-5-0 treatments. In the 5-0-5 treatment, nodule growth was completely inhibited in the first and the third weeks with 5 mm nitrate, but the nodule growth was enhanced in the second week with 0 mm nitrate. The nodule growth response to 5 mm nitrate was similar between small and large size nodules. After the 5-5-5, 5-0-5, 0-0-5, and 0-5-5 treatments, where the plants were cultured with 5 mm nitrate in the last third week, the ARA per plant was significantly lower compared with the 0-0-0 treatment. On the other hand, the ARA after the 5-0-0 and 5-5-0 treatments was relatively higher than that after the 0-0-0 treatment, possibly due to the higher photosynthate supply associated with the vigorous vegetative growth of the plants supplemented with nitrate nitrogen. It is concluded that both soybean nodule growth and N₂ fixation activity sensitively responded to the external nitrate level, and that these parameters were reversibly regulated by the current status of nitrate in the culture solution, possibly through sensing of the nitrate concentration in roots and / or nodules.     

Effect of CO2-free air treatment on nitrogen fixation of soybeans inoculated with Bradyrhizobium japonicum and Rhizobium fredii

Soybean (Glycine max L. Merr.) cultivars Akisirome and Peking were inoculated with Bradyrhizobium japonicum Is-21 and Rhizobium fredii USDA 194, respectively, and were grown in cylindrical pots containing sterilized vermiculite which were aerated with CO₂-free air or ambient air to study the effects of CO₂ deficiency in the rhizosphere on plant growth, nodulation, and nitrogen fixation. The repressive effects of CO₂-free air treatment were more conspicuous in Peking than Akisirome, and nodule number, nodule mass, amount of biologically fixed N and plant growth of Peking were reduced remarkably by the CO₂-free air treatment.

Acetylene reduction activity (ARA) of Peking inoculated with USDA 194 and Akisirome inoculated Is-21 was assayed in the absence and presence of CO₂, ARA of Akisirome was not affected by the absence of CO₂, while that of Peking decreased drastically.

Based on these results, it was concluded that nitrogen fixation by Akisirome inoculated with B. japonicum Is-21 was not appreciably influenced by CO₂ deficiency in the rhizosphere, while that of Peking inoculated with R. fredii USDA 194 was severely repressed, and the decrease was estimated to be due to both the reduction of the nodule mass and specific nitrogen-fixing activity.     

Effect of soil physical properties on soybean nodulation and N2 fixation at the early growth stage in heavy soil field in Hachirougata Polder,Japan

We studied the effect of the soil physical properties on soybean nodulation and N₂ fixation in the heavy soil of an upland field (UF) and an upland field converted from a paddy field (UCPF) in the Hachirougata polder, Japan. Seeds of the soybean cultivar Ryuho were sown in each field with or without inoculation of Bradyrhizobium japonicum A1017. The soybean plants were sampled at 35 (V3) and 65 (Rl) d after sowing (DAS), and then nodulation and the percentage of N derived from N₂ fixation in the xylem sap were determined. The soil physical properties were different between UF and UCPF, especially the air permeability and soil water regime. Nodule growth was restricted in UCPF irrespective of rhizobial inoculation, though rhizobial infection was not inhibited by the unfavorable soil physical conditions. Soybean plant growth was closely related to the nodule mass and N₂ fixation activity, and the inoculation of a superior rhizobium strain was effective only at 35 DAS. These results indicate that soybean nodulation and N₂ fixation was considerably affected by the physical properties of heavy soil, and that it is important to maintain the N₂ fixation activity and inoculate the soybean plants with a superior rhizobium strain at a later growth stage in order to increase soybean production in heavy soil fields.