隔根与接种 FM 对红壤上玉米/大豆植株生长及氮素利用的影响

【目的】探讨接种摩西管柄囊霉 (Funneliformis mosseae,FM) 和不同隔根处理对红壤上间作植株生长、植株氮吸收量和土壤氮的影响。【方法】采用盆栽模拟试验,设不同菌根处理[不接种 (NM)、接种 (FM)]与玉米/大豆不同隔根处理 (根系不分隔、部分分隔、完全分隔)。【结果】接种 FM 的玉米、大豆根系均有一定的侵染,菌根侵染率在部分分隔处理下最低。间作根系的分隔处理对玉米和大豆的菌根依赖性产生了明显影响,大豆的菌根依赖性随间作交互作用强度的加大而增加。无论何种隔根处理,接种 FM 均显著增加了玉米植株生物量,其地上部生物量高出 NM 处理 11.7%～81.4%,根系生物量高出 NM 处理 18.8%～166.7%。根系分隔处理下,接种 FM 均显著降低了大豆生物量。同一隔根方式下,接种 FM 明显提高了玉米的植株氮吸收量和根系氮吸收效率。在不分隔处理下,接种 FM 显著增加了大豆的地上部氮吸收量,但在部分分隔和完全分隔处理下则反而有所下降;在部分分隔处理下,接种 FM 显著降低了大豆根系的氮吸收量,在不分隔和完全分隔处理下亦呈下降趋势。在部分分隔处理下,接种 FM 显著提高了大豆根系氮吸收效率,在完全分隔处理下反而有明显下降,且在 NM–不分隔处理下的大豆根系氮吸收效率最低。相关分析显示,玉米、大豆植株氮吸收量与土壤碱解氮含量呈显著负相关。【结论】接种丛枝菌根真菌 (AMF) 和隔根方式的组合能不同程度地影响玉米和大豆对氮的吸收利用及间作植株的生长,并能对土壤有效氮产生较大影响。所有的复合处理中,AMF和间作根系部分分隔处理组合对玉米和大豆生长及氮素利用的促进作用较好,并能有效降低土壤碱解氮的残留。     

菌根与根瘤菌联合应用对复垦矿区根际土壤环境的改良后效

为了进一步确定微生物复垦对土壤的持续修复能力,在神东矿区活鸡兔开采沉陷试验基地,接种丛枝菌根真菌和大豆根瘤菌改良农用地土壤,已取得了较好的微生物复垦效果,经过2 a的退耕撂荒以后,对微生物复垦后效进行系统地研究。研究结果表明,撂荒后,接菌小区根外菌丝密度、根系侵染率和pH值均显著高于对照小区,分别比对照提高了90%、52%和1.3%;接菌小区细菌数量、微生物总量、有效磷含量、电导率和酸性磷酸酶活性也明显高于对照小区,但差异不显著。撂荒后与撂荒前相比,土壤呈退化趋势,但接菌小区菌丝密度几乎增加了1倍,差异显著,同时侵染率、细菌数量和酸性磷酸酶活性均明显升高,差异不显著;而在对照小区中,菌丝密度稍有增加,侵染率、细菌数量和酸性磷酸酶活性均明显降低,且差异不显著。利用丛枝菌根真菌与大豆根瘤菌联合作为绿色环保肥料,能够有效改善根际土壤环境,减轻土壤的退化程度,在弃耕状态下也能发挥良好的效用,对土地贫瘠的开采沉陷地进行生态修复具有重大意义。     

Field performance of three Bradyrhizobium japonicum strains with two soybean (Glycine max. L.) cultivars

Summary A field experiment was condutced in a clay loam soil to study the performance of three Bradyrhizobium japonicum strains; USDA 110, USDA 138 and TAL 379, in relation to their N₂-fixing potential and competitiveness on two soybean cultivars (Clark and Calland). Inoculation of soybean cultivars with these strains, either singly or in combination, induced significant increases in plant dry weight, N₂ fixation and seed yields. However, no significant differences were found between the rhizobial strains and/or their mixtures in N₂ fixation and increased seed yield for both cultivars. The two soybean cultivars gave similar responses to inoculation. No significant differences in seed yield were observed between Clark and Calland cultivars. The interaction between inoculant strain and soybean cultivar was not significant. The competition between strains for nodulation was assessed. Strain USDA 110 was the most competitive, followed by USDA 138. Strain TAL 379 was always less competitive on both cultivars. The incidence of double-strain occupancy of nodules varied from 8% to 40%.     

Effects of co-inoculation of Bradyrhizobium elkanii BLY3-8 and Streptomyces griseoflavus P4 on Rj4 soybean varieties

Co-inoculation of selected nitrogen-fixing bacteria with plant growth-promoting bacteria is the promising way for the improvement of soybean production through enhancing plant growth, nodulation, and N₂ fixation. Therefore, this experiment was conducted to study the effects of co-inoculation of Bradyrhizobium elkanii BLY3-8 with Streptomyces griseoflavus P4 on plant growth, nodulation, N₂ fixation, N uptake, and seed yield of Rj₄ soybean varieties. Two experiments with completely randomized design and three replicates were done in this study. N₂-fixation ability of soybean was evaluated by acetylene reduction activity (ARA) and relative ureide method. In the first experiment, synergetic effect in N₂ fixation and nodulation was occurred in co-inoculation treatment (BLY3-8 + P4) in Yezin-3 and Fukuyutaka. Based on these results, co-inoculation effect of BLY3-8 and P4 was assessed on Yezin-3 and Fukuyutaka varieties at three different growth stages, using Futsukaichi soil under natural environmental conditions. This study shows that co-inoculation of BLY3-8 and P4 significantly increased N₂ fixation at V6 stage; plant growth, nodulation, N₂ fixation, and N uptake at R3.5 stage; and shoot growth, N uptake, and seed yield at R8 stage, in Rj₄ soybean varieties compared with the control. Significant difference in plant growth, nodulation, N₂ fixation, N uptake, and yield between co-inoculation and control, not between single inoculation and control, suggests that there is a synergetic effect due to co-inoculation of BLY3-8 and P4.     

Regulation of mycorrhiza development in durum wheat by P fertilization: Effect on plant nitrogen metabolism

The aim of this work was to study the effect of arbuscular mycorrhizal fungus Glomus mosseae on growth and nitrogen (N) metabolism of durum wheat (Tritcum durum) under various P soil contents. The analyses were extended to macro and micronutrient tissue concentrations, nitrate reductase and glutamine synthetase activities, as well as protein, aminoacids, pyridine dinucleotides and adenine nucleotides. Arbuscular mycorrhiza increased wheat growth in soil in which P availability was low and nitrate was the dominant N form. The root colonization occurred at the highest level in plants grown in limiting soil P and was inversely related to soil P content. The micorrhizal wheat plants contained also the highest concentrations of macro (P, K, Ca, N) and micronutrients (Fe, Zn, Mn) as well as free amino acids, protein, NAD, NADP, AMP, ADP, ATP in roots and leaves. In particular, the micronutrient tissue concentrations (Zn, Mn) supported that mycorrhiza actively modulated their uptake limiting interferences and optimizing growth better than the plant roots, like a very efficient “rootstock”. Control plants grown at the highest soil P did not reach the same concentration as the mycorrhizal plants. Nitrate reductase activities in the roots of mycorrhizal plants were higher than in the control ones, while glutamine synthetase activities were highest in the leaves. Protein and amino acids concentrations, as well as AMP, ADP, ATP, NAD(P), and NAD(P)H were also higher than in the control. Among the free amino acids in the roots, the high levels of glutamine, asparagine, arginine, support the view that ammonium was transferred through the arbuscules to the root cells where it was re‐assimilated in the cortical cells, forming high N : C ratio‐amino acids. They were transferred to the leaves where all the other N compounds could be largely synthesized using the carbon skeletons supplied by photosynthesis.     

AM真菌菌丝际细菌具有固氮解磷双重功能

为了认识和理解丛枝菌根真菌(AM真菌)根外菌丝表面定殖的细菌是否同时具有固氮和解磷能力,从田间生长的玉米菌根根外菌丝表面分离鉴定了固氮菌,评价了其固氮和解磷能力.从AM真菌根外菌丝表面分离出23株可在无氮培养基中生长的细菌,分属于变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)和放线菌门(Ac...     

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.     

Denitrifying ability of indigenous strains of Bradyrhizobium japonicum isolated from fields under paddy-upland rotation

Summary In Japan some paddy fields are used for upland crops for several years and then returned to paddy fields (paddy-upland rotation). Soybeans (Glycine max L.) are an important summer crop. The ability to denitrify and some characteristics of denitrification by isolated strains of Bradyrhizobium japonicum were investigated to clarify the frequency of denitrifiers in indigenous populations of B. japonicum in fileds under paddyupland rotation. Eight field plots with different cropping systems at two sites were used. The fields consisted of a Gray Lowland Soil, and either soybeans or paddy rice (Oryza sativa L.) was grown as a summer crop, with barley (Hordeum distichum L.) or wheat (Triticum aestivum L.) as a winter crop. All B. japonicum strains present in the plots were able to denitrify. Isolated strains fell into two main groups (groups I and II) according to the rate of denitrification. Strains of group I evolved N₂O with C₂H₂ at a rate comparable to that of Alcaligenes denitrificans IAM 12370, whereas the denitrification activity of group II strains was 100 times lower than that of group I strains. Both group I and II strains occurred in each plot. Amounts of N₂O produced by indigenous strains with and without C₂H₂ suggested that strains of group I and II evolved N₂ or N₂O, respectively, as the end product of denitrification. One strain (S 107) that was isolated had the highest denitrifying ability with an end product of N₂O. These results indicate that indigenous bradyrhizobia may partly contribute to denitrification of field soil under a paddy-upland rotation.     

The effects on N2 fixation (C2H2 reduction), bacterial population and rice plant growth of two modes of straw application to a wetland rice field

Summary The effects of incorporation and surface application of straw to a wetland rice field on nitrogen fixation (C₂H₂ reduction), bacterial population and rice plant growth were studied. Rice straw (5 t ha^–1) was chopped (10- to 15-cm pieces) and applied to the field 2 weeks before transplanting IR42, a long-duration variety, and IR50, a short-duration variety. The acetylene-reducing activity (ARA) of IR42 and IR50 measured at heading stage for 3 consecutive days showed significantly higher ARA in IR42 as a result of the 2 straw application methods. Mostly up to 20 days after straw surface application and incorporation, the dark ARA in the soil, total and N₂-fixing heterotrophs, and photoorganotrophic purple nonsulphur bacteria (POPNS) in the soil and in association with degrading straw were stimulated. Higher bacterial populations were associated with straw on the surface than with straw incorporated. The POPNS counts, in particular, were increased hundreds fold in the surface-applied straw treatment. Straw applications also increased the root, shoot and total plant biomass at heading stage and the total dry matter yield at harvest in both varieties. The data show the potentials of straw as a source of substrate for the production of microbial biomass and for the non-symbiotic N₂ fixation to improve soil fertility and plant nutrition.     

三种土壤上六种丛枝菌根真菌生长特征和接种效应

以分离于华北、华中和华南3个生态区及法国引进的丛枝菌根真菌为试验菌株,采用三室根箱培养的方法,研究了它们在华北、华中和华南3种典型土壤褐土、棕壤和红壤上的菌根形成、接种效应、磷吸收贡献和根外菌物量情况。结果表明,6种菌株在上述指标上存在显著的种间或生态型差异,土壤与菌株间存在显著的交互作用。6种菌株在3种土壤上都能与玉米形成菌根,在褐土和棕壤上大多数菌株在分离地所在地区土壤类型上的菌根侵染率较高,说明其对该土壤条件的适应性较强;在红壤强酸性土壤条件抑制了菌根真菌的侵染。菌株BEG168、BEG167、BEG151、BEG221和BEG141在褐土上,BEG151和BEG221在棕壤上,BEG168和BEG150在红壤上显著提高了玉米的生物量。在褐土和棕壤上,除BEG150外,BEG168、BEG167、BEG151、BEG221和BEG141能显著促进宿主吸磷;而在红壤上,BEG168和BEG141显著促进了宿主吸磷。若以真菌的根外菌物量作为衡量AM真菌菌株土壤生态适应性的指标,BEG141和BEG167是生态适应性强的菌株,为广幅生态型菌株;菌株BEG168和BEG151次之,前者在棕壤上适应性高,后者在红壤上适应性高。BEG150和BEG221生态适应性较窄,仅适应红壤或褐土,为窄幅生态型菌株。Glomus.etunicatum的两个生态型BEG168和BEG221在土壤生态适应性上差异很大,前者在两种土壤上收集到菌物量,而后者只在褐土上收集到菌物量。土壤条件可以决定丛枝菌根真菌的生长状况和功能。     

Competition among strains of Bradyrhizobium and vesicular-arbuscular mycorrhizae for groundnut (Arachis hypogaea L.) root infection and their effect on plant growth and yield

Summary Strains of Bradyrhizobium influenced root colonization by a species of vesicular-arbuscular mycorrhizae (VAM), and species of VAM influenced root nodulation by strains of Bradyrhizobium in pot experiments. In a field experiment, the effects of VAM on competition amongst inoculated bradyrhizobia were less evident, but inoculation with Bradyrhizobium strains increased root colonization by VAM. Certain VAM/Bradyrhizobium inoculum strain combinations produced higher nodule numbers. Plants grown without Bradyrhizobium and VAM, but supplied with ammonium nitrate (300 g ml^–1) and potassium phosphate (16 g ml^–1), produced higher dry-matter yields than those inoculated with both symbionts in the pot experiment. Inoculation with either symbiont in the field did not result in higher pod and haulm yields at harvest.ICRISAT Journal Article No. 886     

Nickel-tolerant Brevibacillus brevis and arbuscular mycorrhizal fungus can reduce metal acquisition and nickel toxicity effects in plant growing in nickel supplemented soil

The growth of clover (Trifolium repens ) and its uptake of N, P and Ni were studied following inoculation of soil with Rhizobium trifolii, and combinations of two Ni-adapted indigenous bacterial isolates (one of them was Brevibacillus brevis) and an arbuscular mycorrhizal (AM) fungus (Glomus mosseae). Plant growth was measured in a pot experiment containing soil spiked with 30 (Ni I), 90 (Ni II) or 270 (Ni III) mg kg⁻¹ Ni-sulphate (corresponding to 11.7, 27.6 and 65.8 mg kg⁻¹ available Ni on a dry soil basis). Single inoculation with the most Ni-tolerant bacterial isolate (Brevibacillus brevis) was particularly effective in increasing shoot and root biomass at the three levels of Ni contamination in comparison with the other indigenous bacterial inoculated or control plants. Single colonisation of G. mosseae enhanced by 3 fold (Ni I), by 2.4 fold (Ni II) and by 2.2 fold (Ni III) T. repens dry weight and P-content of the shoots increased by 9.8 fold (Ni I), by 9.9 fold (Ni II) and by 5.1 fold (Ni III) concomitantly with a reduction in Ni concentration in the shoot compared with non-treated plants. Coinoculation of G. mosseae and the Ni-tolerant bacterial strain (B. brevis) achieved the highest plant dry biomass (shoot and root) and N and P content and the lowest Ni shoot concentration. Dual inoculation with the most Ni-tolerant autochthonous microorganisms (B. brevis and G. mosseae) increased shoot and root plant biomass and subtantially reduced the specific absorption rate (defined as the amount of metal absorbed per unit of root biomass) for nickel in comparison with plants grown in soil inoculated only with G. mosseae. B. brevis increased nodule number that was highly depressed in Ni I added soil or supressed in Ni II and Ni III supplemented soil. These results suggest that selected bacterial inoculation improved the mycorrhizal benefit in nutrients uptake and in decreasing Ni toxicity. Inoculation of adapted beneficial microorganisms (as autochthonous B. brevis and G. mosseae) may be used as a tool to enhance plant performance in soil contaminated with Ni.     

Changes in acetylene reduction activities and effects of inoculated rhizosphere nitrogen-fixing bacteria on rice

Acetylene reduction activities (ARAs) of soils and rice plants during rice-growing season were monitored in temperate region in northeast China. This activity was significantly higher in rhizosphere soil than that in inter-row soil after rice seedlings were transplanted. The ARA was high for most of growing season, suggesting that the native N₂-fixing bacteria responded to rice roots very quickly. Sixteen strains of free-living N₂-fixing bacteria were isolated from three different soils. The ARAs of these strains were correlated with the averaged soil ARAs, suggesting that the isolated strains were likely the active flora responsive to rice roots. The strains were inoculated by soaking seedling roots into the liquid culture for 2 h, and the seedlings were transplanted into pots. Most strains tested did not show any growth-promoting effects except Azotobacter armeniacus and Azotobacter nigricans, which showed growth-promoting effects only at late rice growth stage and only when inoculated in combination but not separately. Present data indicated the promising future applications of these two strains in combination in the region, but further research is needed to understand the underlying mechanisms.     

Effects of deep placement of controlled-release nitrogen fertilizer on soybean growth and yield under sulfur deficiency

Sulfur (S) and Nitrogen (N) metabolisms in plants are interacted and it is known that S deficiency decrease N absorption and metabolism. In leguminous plants S deficiency also decreases N₂ fixation by rhizobia in the nodules. Deep placement of a controlled-release N fertilizer is a good method to provide nitrogen to soybean without inhibiting N₂ fixation; thus, it was hypothesized that this method is able to provide nitrogen effectively to sulfur-deficient soybean plants. In this study effects of deep placement of coated urea on S-N physicological interaction, growth and productivity in soybean plants were examined using pot experiments. Soybean plants were grown with sulfate concentrations of 30, 100, or 1000 μM, with or without deep placement of coated urea. Shoot weights at the developing stage were not affected by S deficiency. SPAD values of leaves during the flowering stage decreased with S deficiency and increased with the deep placement of coated urea. S deficiency decreased seed weight per plant at the harvesting stage, but this decrease was attenuated by the deep placement of coated urea. N and S content in shoots at the developing stage increased with the deep placement of coated urea, whereas in seeds, only the N content increased. N₂ fixation activity based on the relative ureide-N content in xylem sap indicated that the deep placement of coated urea did not inhibit N₂ fixation activity at the early flowering stage. Without deep placement of coated urea, the relative ureide-N content decreased under S deficiency at the seed filling stage. These results suggest that the deep placement of coated urea is an efficient method to supply N to support soybean yield under S deficiency.

Abbreviations: Deep+: with deep placement of coated urea; Deep–: without deep placement of coated urea     

Nitrogen fixation and soil N level during maturation affect the contents of storage compounds of soybean seeds

We compared the concentrations and contents of protein and oil in mature seeds from nodulated and non-nodulated soybean plants grown on soils with four different N levels during maturation. We observed a positive correlation between the contents of protein and oil in seeds from nodulated plants. Seeds from nodulated plants grown on urea-treated soil showed higher protein and lower oil contents than those from plants grown on soil treated with coated slow release N fertilizer (LP-100). Contents of these compounds in seeds from nodulated plants grown on LP-100 soil were almost the same as those from non-nodulated plants on the same soil. These observations indicated that N economy in roots during seed maturation affects the contents of storage compounds. We suggested that the control of the N₂ fixation activity of soybean plants and management of soil N level during seed maturation are important to determine the contents of protein and oil in seeds.     

Integration of arbuscular mycorrhiza inoculation in hydroseeding technology: effects on plant growth and inter‐species competition

Hydroseeding is a technique increasingly used to establish vegetation on large degraded areas, such as large‐scale road construction sites and quarries. Native grasses and legume species are used on rehabilitation and restoration projects as a first step in the recovery of such places, prior to the establishment of native forbs and shrubs that occurs at a slower pace. The effect of mycorrhizal inoculation on the development of nine species of grasses and legumes that can be potentially used in restoration processes in the Mediterranean area was studied, in microcosm experiments under greenhouse conditions. The effect of adding arbuscular mycorrhizal (AM) inoculum to a hydroseeding mixture was also investigated in greenhouse and in field conditions. In the hydroseeding experiments the mycorrhizal inoculum was added to the seed slurry in a jet agitated hydroseeding machine and sprayed to the soil surface with a pressurised spray in a one‐step application. The study shows that Glomus intraradices Schenk & Smith BEG72 is able to establish the symbiosis when applied at sowing while Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe BEG116 is not. It also confirms that legumes are more highly mycotrophic than grasses. The results of the hydroseeding experiments demonstrate the establishment of the symbiosis using this technology, both in the greenhouse and in the field. Mycorrhizal inoculation improved above ground plant growth and increased the legumes/grasses ratio. Copyright © 2007 John Wiley & Sons, Ltd.     

Copper,nitrogen, and Rhizobium japonicum relationships in determinate soybean

A relationship among Cu, N, and Rhizobium japonicum was hypothesized because previous research had shown that (a) 35% or more legumes in the Atlantic Coastal Plain have Cu concentrations of 6 mg kg^‐1 or less, (b) Cu influences N fixation in some legumes, and (c) irrigated soybean (Glycine max L. Merr.) can accumulate most of its N through fixation. Soybean were grown on a Cu‐deficient Norfolk (fine‐loamy, siliceous, thermic Typic Paleudult) loamy sand with 3 fertilizer sources of Cu, 2 strains of R. japonicum, and with or without 336 kg ha^‐1 of N fertilizer. Application of Cu significantly increased the number of pods plant^‐1 suggesting pod abortion in determinate soybean may be caused by low Cu, but seed yield was not increased. Fertilization with N increased vegetative growth, but not total biomass or seed yield. Inoculation with R. japonicum strain 110 significantly increased seed yield by 0.3 Mg ha^‐1 compared to strain 587. The yield increase was similar with or without fertilizer N application indicating strain response was not totally caused by improved N efficiency. There was no relationship between seed yield and nodule occupancy as measured by the ELISA technique.     

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.     

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.