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.     

Yield,nodulation, and N2 fixation by cowpea cultivars at different phosphorus levels

A greenhouse experiment was conducted to investigate the effect of a P application (0 vs. 50 mg P kg^-1) on yield, nodulation, and N₂ fixation by three cowpea cultivars (Soronko, Amantin, and IT81D-1137) using the ¹⁵N isotope-dilution method. When P was not applied the inoculated cowpea genotypes showed significant differences (Soronko>Amantin> IT81D-1137) in N accumulation, in contrast to the uninoculated cowpea cultivars, which accumulated similar amounts of N. The differences in shoot N in inoculated plants were thus caused by differences in N₂ fixation. The average values of N fixed (for both P levels) were 74% in Soronko, 59% in Amantin, and 42% in IT81D-1137, corresponding to 80, 51, and 24 mg N plant^-1, respectively. Inoculation increased the total shoot-N accumulation in cv. Soronko by 270% without P and by 204% with P, cv. Amantin by 152 and 104%, and cv. IT81D-1137 by 74 and 58%, respectively. With P, the % N derived from atmosphere (%Ndfa) was 42% for IT81D-1137, 62% for Amantin, and 76% for Soronko. The high value for Soronko indicates that in a soil of medium fertility, certain cowpea cultivars are capable of satisfying their total N requirement through N₂ fixation. The P effect on N₂ fixation was mainly in the total amount of N fixed rather than on the percentage derived from the atmosphere.     

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.     

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.     

Application of N2/Ar ratio method for N2 fixation studies in submerged soil

Abstract

Recently there has been developments in the measurement of N₂ fixation due mainly to the C₂H₂ reduction method (1). This method, however, has several disadvantages, especially for submerged soil, and the estimated amount of fixed N₂ on the basis of the C₂H₂ reduction activity is not very reliable. The tracer ¹⁵N₂ technique which gives a reliable estimation of the fixed N₂ is too expensive for common use. Development of an alternative method suitable for submerged soil would therefore be desirable. The present authors expected that the measurement of the ratio N₂/Ar in the soil solution might provide advantages for the estimation of the fixed N₂ in submerged soil.     

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.     

Effects of Rhizobium and vesicular-arbuscular mycorrhiza inoculations on nodulation,symbiotic nitrogen fixation and soybean yield in subtropical-tropical fields

Summary Field experiments were carried out to determine the effects of single and mixed inoculations with Rhizobium and vesicular-arbuscular mycorrhiza (VAM) on nodulation, symbiotic N₂ fixation and yield of soybeans in six Taiwan subtropical-tropical sites. Inoculation with Rhizobium alone significantly increased nodulation, nodule weight and nitrogenase activity of nodules in three out of six experimental fields, and affected soybean yields in the range –13% to + 134%. Inoculation with VAM fungi alone did not have a significant effect on nodulation and nitrogenase activity. Mycorrhiza inoculation affected soybean yields in the range –13% to + 65%, but only the yield increases at one out of six sites with N application were statistically significant. Mixed inoculation with Rhizobium and mycorrhiza affected yields in the range –8% to + 145% A synergistic effect from mixed inoculation of Rhizobium-mycorrhiza on soybean yields was found in one out of six experimental fields. The yield response to N application (40 kg N ha^–1) in these six paddy-field trials was not significant. These results suggest that single or mixed inoculation of rhizobia can greatly assist soybean grain yields and can replace N fertilizers.     

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.     

壳聚糖对NaCl胁迫下菜用大豆结瘤固氮的影响

【目的】研究壳聚糖对盐胁迫抑制菜用大豆结瘤固氮的缓解效应,为进一步探讨壳聚糖抗逆机理提供新的线索。【方法】以蛭石为基质,以菜用大豆‘特早王’–根瘤菌共生体系为研究对象,采用人工气候箱培养,研究NaCl胁迫下壳聚糖对菜用大豆根瘤形成、生物固氮的影响。菌种为与‘特早王’共生匹配性较好的快生根瘤菌N18。接种后的植株进行如下4个处理:1)叶面喷施清水,根部浇灌无氮营养液(CK);2)叶面喷施壳聚糖水溶液,根部浇灌无氮营养液(CTS);3)叶面喷施清水,根部浇灌溶有NaCl的无氮营养液(Cl);4)叶面喷施壳聚糖水溶液,根部浇灌溶有NaCl的无氮营养液(CTS+Cl)。上述各处理施用的水或水溶液均为无菌水配制,NaCl处理的浓度为50 mol/L,CTS处理的适宜浓度为200 mg/L。接种30天后,将大豆植株取出,用清水将根部蛭石冲洗干净后,立即测定根瘤固氮酶活性、根瘤数及根瘤鲜重,然后测定根瘤豆血红蛋白含量和根系活力,最后测植株干重和全氮量。【结果】氯化钠胁迫下,植株干重显著下降,与CK相比降幅达49%,喷施壳聚糖后(CTS+Cl),降低幅度显著减小,但依然显著低于CK (P <0.05)。无盐条件下,与CK相比,壳聚糖处理(CTS)增加植株干重的效果不明显。喷施壳聚糖显著增加了菜用大豆的根瘤数、根瘤鲜重、植株含氮量、根系活力、豆血红蛋白含量及固氮酶活性(P <0.05)。NaCl胁迫显著抑制了菜用大豆的结瘤固氮作用,其中根瘤数、根瘤鲜重分别较CK下降了79%、90%,而壳聚糖处理(CTS+Cl)使菜用大豆在盐逆境下的结瘤数、根瘤鲜重、植株全氮含量、根系活力、豆血红蛋白含量及固氮酶活性等均显著回升,增幅分别达对照的29%、20%、17%、48%、19%、21%,但均显著低于CK。【结论】非NaCl胁迫下,喷施壳聚糖可以显著促进菜用大豆结瘤,提高豆血红蛋白含量及固氮酶活性,最终增加植株含氮量。在NaCl胁迫下,外源壳聚糖可以显著缓解氯化钠胁迫导致的对根系活力和结瘤固氮的影响。因此,叶面喷施壳聚糖是促进菜用大豆结瘤固氮和生长的有效措施。     

Elevated CO2 concentrations increase leaf nitrate reduction by strengthening sink activity in soybean plants

An experiment was conducted to examine the effect of CO₂ enrichment on the nitrate uptake, nitrate reduction activity, and translocation of assimilated-N from leaves at varying levels of nitrogen nutrition in soybean using ¹⁵N tracer technique. CO₂ enrichment significantly increased the plant biomass, apparent leaf photosynthesis, sugar and starch contents of leaves, and reduced-N contents of the plant organs only when the plants were grown at high levels of nitrogen. A high supply of nitrogen enhanced plant growth and increased the reduced-N content of the plant organs, but its effect on the carbohydrate contents and photosynthetic rate were not significant. However, the combination of high CO₂ and high nitrogen levels led to an additive effect on all these parameters. The nitrate reductase activity increased temporarily for a short period of time by CO₂ enrichment and high nitrogen levels. ¹⁵N tracer studies indicated that the increase in the amount of reduced-N by CO₂ enrichment was derived from nitrate-N and not from fixed-N of the plant. To examine the translocation of reduced-N from the leaf in more detail, another experiment was conducted by feeding the plants with ¹⁵NO₃-N through a terminal leaflet of an upper trifoliated leaf under depodding and/or CO₂ enrichment conditions. The export rate of ¹⁵N from the terminal leaflet to other plant parts decreased by depodding, but it increased by CO₂ enrichment. CO₂ enrichment increased the percentage of plant ¹⁵N in the stem and / or pods. Depodding increased the percentage of plant ¹⁵N in the leaf and stem. The results suggested that the increase in the leaf nitrate reduction activity by CO₂ enrichment was due to the increase of the translocation of reduced-N from leaves through the strengthening of the sink activity of pods and / or stem for reduced-N.     

Effect of soybean inoculation with Bradyrhizobium and wheat inoculation with Azotobacter on their productivity and N turnover in a Vertisol

A long-term field experiment was conducted for 8 years on a Vertisol in central India to assess quantitatively the direct and residual N effects of soybean inoculation with Bradyrhizobium and wheat inoculation with Azotobacter in a soybean–wheat rotation. After cultivation of soybean each year, its aerial residues were removed before growing wheat in the same plots using four N levels (120, 90, 60 and 30 kg ha^?1) and Azotobacter inoculation. Inoculation of soybean increased grain yield by 10.1% (180 kg ha^?1), but the increase in wheat yields with inoculation was only marginal (5.6%; 278 kg ha^?1). There was always a positive balance of soil N after soybean harvest; an average of +28 kg N ha^?1 yr^?1 in control (nodulated by native rhizobia) plots compared with +41 kg N ha^?1 yr^?1 in Rhizobium-inoculated plots. Residual and direct effects of Rhizobium and Azotobacter inoculants caused a fertilizer N credit of 30 kg ha^?1 in wheat. Application of fertilizers or microbial inoculation favoured the proliferation of rhizobia in crop rhizosphere due to better plant growth. Additional N uptake by inoculation was 14.9 kg N ha^?1 by soybean and 20.9 kg N ha^?1 by wheat crop, and a gain of +38.0 kg N ha^?1 yr^?1 to the 0–15 cm soil layer was measured after harvest of wheat. So, total N contribution to crops and soil due to the inoculants was 73.8 kg N ha^?1 yr^?1 after one soybean–wheat rotation. There was a total N benefit of 13.8 kg N ha^?1 yr^?1 to the soil due to regular long-term use of microbial inoculants in soybean–wheat rotation.     

Effect of nodulation with Bradyrhizobium japonicum and Shinorhizobium fredii on xylem sap composition of Peking (Glycine max L. Merr )

Five barley cultivars were grown together in complete, low-P·low-pH and high-Al medium containing only NO₃, only NH₄ or both NO₃ and NH₄ as N sources, respectively using an automatic control system of pH for water culture, and the relationship between the differential Al tolerance and the plant-induced pH change of medium among the barley cultivars was investigated.

The pH of the medium containing only NO₃ as N source tended to increase, whereas the pH of the other media containing only NH₄ or both NO₃ and NH₄ as N sources tended to decrease, but the fluctuations of the medium pH could be maintained within the value of 0.2 pH in the complete medium and within the value of 0.1 pH in the high-Al medium.

Barley cultivars still differed in their Al tolerance in the medium which was continuously stirred and circulated at a constant pH. The pattern of Al tolerance was not affected by the N sources in the medium. The plant-induced pH change of medium for each cultivar was influenced by the N sources in the medium, and was not correlated positively with Al tolerance. The contents of Al and Ca or other nutrient cations in roots were positively correlated with Al tolerance and positive correlations were recognized also between the contents of Al and Ca or some other nutrient cations in the roots.

In conclusion, the following mechanisms are proposed. Al tolerant barley cultivars exclude Al actively outside the plasmalemma of the root cells, and the excluded Al may polymerize and or react with P to form Al precipitates. Consequently, in the Al tolerant barley cultivars the Al content may be low in the root protoplasts, high in the whole root tissues and the contents of Ca or other nutrients may be high in the roots. The plant-induced pH change of medium is not considered to be the cause of the differential Al tolerance among barley cultivars.     

Influence of mycorrhiza vs. soluble phosphate on growth,nodulation, and N2 fixation (15N) in alfalfa under different levels of water potential

Summary The legume Medicago sativa (+Rhizobium melilott) was grown under controlled conditions to study the interactions between soluble P in soil (four levels), or a mycorrhizal inoculum, and the degree of water potential (four levels) in relation to plant development and N₂ fixation. ¹⁵N-labelled ammonium sulphate was added to each pot for a qualitative estimate of N₂ fixation, in order to rank the effects of the different treatments.Dry-matter yield, nutrient content and nodulation increased with the amount of plant-available P in the soil, and decreased as the water stress increased, for each P-level. The mycorrhizal effect on dry matter, N yield, and on nodulation was little affected by the water potential. Since P uptake was affected by the water content in mycorrhizal plants, additional mechanisms, other than those mediated by P, must be involved in the mycorrhizal activity.There was a positive correlation between N yield and nodulation for the different P levels and the mycorrhizal treatment at all water levels. A high correlation between plant unlabelled N content and atom% ¹⁵N excess was also found for all levels of P. In mycorrhizal plants, however, the correlation between unlabelled N yield and ¹⁵N was lower. This suggests that mycorrhiza supply plants with other N sources in addition to those derived from the improvement on N₂ fixation.     

Microbial activity and bacterial composition of H2-treated soils with net CO2 fixation

The effects of H₂ gas treatment of an agricultural soil cultivated previously with a mixture of clover (Trifolium pratense) and alfalfa (Medicago sativa) on CO₂ dynamics and microbial activity and composition were analyzed. The H₂ emission rate of 250 nmol H₂ g⁻¹ soil h⁻¹ was similar to the upper limit of estimated H₂ amounts emitted from N₂ fixing nodules into the surrounding soil ([Dong, Z., Layzell, D.B., 2001. H₂ oxidation, O₂ uptake and CO₂ fixation in hydrogen treated soil. Plant and Soil 229, 1-12.]). After 1 week of H₂ supply to soil samples simultaneously with H₂ uptake net CO₂ production declined continuously and this finally led to a net CO₂ fixation rate in the H₂-treated soil of 8 nmol CO₂ g⁻¹ soil h⁻¹. The time course of H₂ uptake and CO₂ fixation in the soils corresponded with an increase in microbial activity and biomass of the H₂-treated soil determined by microcalorimetric measurements, fluorescence in situ hybridization analysis (FISH) and DNA staining (DAPI). Shifts in the bacterial community structure caused by the supply of H₂ were recorded. While the H₂ treatment stimulated β-and γ-subclasses of Proteobacteria, it had no significant effect on α-Proteobacteria. In addition, FISH-detectable bacteria of the Cytophaga-Flavobacterium-Bacteroides phylum increased in numbers.     

Effect of CO2 enrichment on carbon and nitrogen interaction in wheat and soybean

Effect of CO₂ enrichment on the carbon-nitrogen balance in whole plant and the acclimation of photosynthesis was studied in wheat (spring wheat) and soybean (A62-1 [nodulated] and A62-2 [non-nodulated]) with a combination of two nitrogen application rates (0 g N land area m^-2 and 30 g N land area m^-2) and two temperature treatments (30/20°C (day/night) and 26/16°C). Results were as follows.

1. Carbon (dry matter)-nitrogen balance of whole plant throughout growth was remarkably different between wheat and soybean, as follows: 1) in wheat, the relationship between the amount of dry matter (DMt) and amount of nitrogen absorbed (Nt) in whole plant was expressed by an exponential regression, in which the regression coefficient was affected by only the nitrogen application rate, and not by CO₂ and temperature treatments, and 2) in soybean the DMt-Nt relationship was basically expressed by a linear regression, in which the regression coefficient was only slightly affected by the nitrogen treatment (at 0N, DMt-Nt balance finally converged to a linear regression). Thus, carbon-nitrogen interaction in wheat was strongly affected by the underground environment (nitrogen nutrition), but not by the above ground environment (CO₂ enrichment and temperature), while that in soybean was less affected by both under and above ground environments.

2. The photosynthetic response curve to CO₂ concentration in wheat and soybean was less affected by the CO₂ enrichment treatment, while that in wheat and soybean (A62-2) was affected by the nitrogen treatment, indicating that nitrogen nutrition is a more important factor for the regulation of photosynthesis regardless of the CO₂ enrichment.

3. Carbon isotope discrimination (..:1) in soybean was similar to that in wheat under ambient CO₂, while lower than that in wheat under CO₂ enrichment, suggesting that the carbon metabolism is considerably different between wheat and soybean under the CO₂ enrichment conditions.     

利用15N2直接标记法研究水稻种植对稻田固氮量和固氮活性的影响

稻田固氮对土壤维持肥力有着重要的作用,但水稻种植与固氮菌及其活性之间的关系尚不清楚。本试验利用¹⁵N₂直接标记法测定了下位砂姜土发育的简育水耕人为土在种水稻和不种水稻条件下的生物固氮量,及其在土壤不同层次(0～1、1～5、5～15 cm)和水稻中的分配,并通过实时荧光定量PCR技术测定了土壤中固氮菌nifH DNA及RNA基因数量。结果表明:种水稻处理显著提高了土壤各层固氮量,尤其提高了1～5 cm和5～15 cm土层土壤固氮量对总固氮量的贡献;种水稻处理的总固氮量是不种水稻处理的10.3倍;水稻植株中生物固定的氮占总固氮量的31.48%;在0～1 cm土层,种水稻处理显著提高了nifH RNA基因数量,而对nifH DNA基因数量的增加不显著。可见,水稻种植没有增加固氮菌的数量,稻田固氮量的增加是因为水稻种植极大地促进了固氮菌nifH基因的表达,提高了固氮菌的固氮活性。     

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.     

Evaluation of N2 fixation by applying 15N labeled plant material and ammonium sulfate

Effect of different ¹⁵N labeled sources on the estimation of N₂ fixation was investigated. The combination of ¹⁵N labeled ammonium sulfate, ¹⁵N labeled plant material, and ¹⁵N labeled ammonium sulfate with unlabeled plant material, was examined in pot experiments. Two cultivars of soybean (Glycine max) and one of mungbean (Vigna radiata) were used. No significant difference was observed among the treatments for the estimation of N₂ fixation. This was due to the homogeneity and stability of the ¹⁵N abundance in soil which resulted in a similar N uptake from the soil by the N² fixing and reference crops. The plant yield, total N uptake and amount of N₂ fixed were higher in the Yellow Soil than in the Andosol. The amount of N₂ fixed was strongly influenced by the plant growth and consequently it affected the plant yield. The slow decomposition of plant material in the Andosol resulted in a low yield in both the N₂ fixing and reference crops. Thus, the artificial decrease of the available N content in soil, by application of plant material, did not stimulate N, fixation but suppressed plant growth and N₂ fixation.     

Effect of CO2 enrichment on biomass production,photosynthesis, and sink activity in Soybean cv. Bragg and its supernodulating mutant nts 1007

Soybean (Glycine max L. Merr.) cv. Bragg and its supernodulating mutant nts 1007 were grown in pots containing vermiculite with a N-free nutrient solution in order to examine the effect of elevated CO₂ concentration (100+20 Pa CO₂ ) on biomass production, photosynthesis, and biological nitrogen fixation. The whole plant weight increase in Bragg was higher than in the mutant at a high CO₂ concentration. Apparent photosynthetic activities of the upper leaves in both Bragg and the mutant increased up to 14 d after treatment initiation by the CO₂ enrichment and thereafter decreased to some extent. Both leaf area and leaf thickness of Bragg increased more than in nts 1007. With the elevated CO₂ concentration, biological nitrogen fixation (BNF) also responded in the same manner as biomass production in both Bragg and nts 1007. The increase of BNF in Bragg was largely due to an increase in nodule weight. Starch contents in the leaves of both Bragg and the mutant increased significantly by CO₂ enrichment, with a higher increase in Bragg than in its mutant. Sugar content in leaf differed only slightly in both Bragg and the mutant. N content in leaf decreased in both Bragg and its mutant, with the decrease being more pronounced in Bragg. However, in other plant parts (roots, stem, and petiole + pods), N content increased in the mutant while in Bragg, it decreased in the pod. N accumulation rate was higher in Bragg than in the mutant and increased more in Bragg than in the mutant by CO₂ enrichment. The ureide content in leaf decreased in Bragg but increased in the mutant by elevated CO₂ concentration. In the nodules, ureide content increased in both Bragg and the mutant by CO₂ enrichment. Based on these results, it is suggested that in terms of biomass production and photosynthetic rate, Bragg responded more to elevated CO₂ concentration than its mutant nts 1007. The alleviation of the stunted vegetative growth of the mutant by CO₂ enrichment was limited despite the significant increase in the photosynthetic activity, presumably due to the limitation of sink activity in the growing parts and not to insufficient supply of N through BNF.     

A simple gas chromatographic approach to evaluate CO2 release,N2O evolution,and O2 uptake from soil

Summary We have developed a simple method for the determination of gaseous compounds that reflect microbial activity in soil, as affected by factors such as the presence of an organic amendment (peat) or a variation in soil moisture. The method is based on a gas chromatographic analysis of the headspace of vials containing the soil under examination. A single gas chromatograph can detect up to 10 different gases. As expected, after peat was added to the soil, CO₂ evolution and O₂ uptake increased significantly. Positive relationships were found between the evolution of N₂O, and soil moisture and the amount of peat added to the soil. Both the these variables influenced the CO₂:O₂ ratio. The results given by this method show high reproducibility.