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.     

Soybean nodulation and nitrogen fixation on soil amended with plant residues

Residues from some tree species may contain allelopathic chemicals that have the potential to inhibit plant growth and symbiotic N₂-fixing microorganisms. Soybean [Glycine max (L.) Merr] was grown in pots to compare nodulation and N₂-fixation responses of the following soil amendments: control soil, leaf compost, red oak (Quercus rubra L.) leaves, sugar maple (Acer saccharum Marsh) leaves, sycamore (Platanus occidentalis L.) leaves, black walnut (Juglans nigra L.) leaves, rye (Secale cereale L.) straw, and corn (Zea mays L.) stover. Freshly fallen leaves were collected from urban shade trees. Soil was amended with 20 g kg^-1 air-dried, ground plant materials. Nodulating and nonnodulating isolines of Clark soybean were grown to the R2 stage to determine N₂-fixation by the difference method. Although nodulation was not adversely affected, soybean grown on leaf-amended soil exhibited temporary N deficiency until nodulation. Nodule number was increased by more than 40% for soybean grown on amended soil, but nodule dry matter per plant generally was not changed compared with control soil. Nonnodulating plants were severely N deficient and stunted as a consequence of N immobilization. Nodulating soybean plants grown on leaf or crop residue amended soil were more dependent on symbiotically fixed N and had lower dry matter yields than the controls. When leaves were composted, the problem of N immobilization was avoided and dry matter yield was not reduced. No indication of an allelopathic inhibition on nodulation or N₂-fixation from heavy application of oak, maple, sycamore, or walnut leaves to soil was observed.     

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.     

The effects of rice (Oryza sativa L. ssp. japonica) husk biochar on nitrogen dynamics during the decomposition of hairy vetch in two soils under high-soil moisture condition

ABSTRACT

Legumes, including hairy vetch (Vicia villosa Roth), are widely used as green manures. They fix nitrogen (N) and provide the N to other crops when they decompose, and thus are considered alternatives for chemical N fertilizers. However, N-rich plant residues, including hairy vetch, are also sources of soil nitrous oxide (N₂O) emissions, a greenhouse gas. On one hand, rice (Oryza sativa L. ssp. japonica) husk biochar is widely used as a soil conditioner in Japan and has been reported as a tool to mitigate soil N₂O emissions. We conducted a soil core incubation experiment (1.5 months) to compare the N₂O emissions during the decomposition of surface-applied hairy vetch (0.8 kg dried hairy vetch m^?2 soil) under semi-saturated soil moisture conditions (~100% water-filled pore space (WFPS)), using two soil types, namely Andosol and Fluvisol. Throughout the incubation period, the use of biochar suppressed soil NH₄⁺-N concentrations in Andosol, whereas the effect of biochar on NH₄⁺-N was not clear in Fluvisol. Biochar increased the nitrate (NO₃^?-N) levels both in Andosol and Fluvisol, suggesting a negative influence on denitrification and/or a positive influence on nitrification. Biochar application did not influence the cumulative N₂O emissions. Our study suggests that rice husk biochar is not a good option to mitigate N₂O emissions during the decomposition of surface-applied hairy vetch, although this study was performed under laboratory conditions without plants. However, the trends of the inorganic-N concentration changes followed by the addition of hairy vetch and biochar were markedly different between the two soil types. Thus, factors behind the differences need to be further studied.     

Effects of symbiosis with Rhizobium fredii on transport of fixed nitrogen in the xylem of soybean plants

An experiment was conducted to identify the main nitrogenous compound transported in the xylem sap of soybean plants nodulated with Rhizobium fredii. Soybean (Glycine max L. Merr.) cultivars, wild type Bragg (nod⁺, fix⁺) and its nitrate tolerant, hypernodulating mutant ntsll16 (nod⁺⁺, fix⁺) were used for this experiment. These soybean plants were inoculated with a slowgrowing rhizobium, Bradyrhizobium japonicum USDAllO or fast-growing rhizobia consisting of a mixture of R. fredii USDA191, USDA193, and USDA-194 and grown in a phytotron under natural light and controlled temperature conditions. Xylem sap was collected from Bragg and ntsll16 plants at the flowering and pod elongation stages. Acetylene reduction activity per plant or per nodule weight was not different between soybean lines and inoculums. The composition of the nitrogenous compounds in the xylem sap was compared between the symbionts, with B. japonicum and R. fredii. At the flowering stage, ureide-N and amide-N accounted for 53 to 70% and 20 to 27% respectively of the total N in the sap collected from the plants inoculated either with B. japonicum or R. fredii. At the pod elongation stage, ureide-N and amide-N accounted for 74 to 85%, and 7 to 19% of total sap N. With the growth of the soybean plants, the ratio of ureide-N in the xylem sap increased. These results suggest that in the case of wild soybean and the hypernodulating mutant line nodulated by R. fredii, ureide is transported as the main nitrogenous compound of fixed nitrogen in the xylem sap in the same way as in plants nodulated with B. japonicum.     

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 CO2 concentration in rhizosphere on nodulation and N2 fixation of soybean and cowpea

Soybean (Glycine max L. Merr.) cvs. Akisengoku and Peking, and cowpea (Vigna unguiculata Walp.) cv. Kegonnotaki were inoculated with Bradyrhizobium japonicum AlO17, Shinorhizobium fredii USDAI93, and B. sp. Vigna MAFF03-03063, respectively and were cultured hydroponically with supply of CO₂-free air, 3dm³ m^-3 CO₂ air, or 25 dm³ m^-3 CO₂ air to study the effects of the CO₂ concentration in the rhizosphere on plant growth, nodulation, and nitrogen fixation. Increase of the CO₂ concentration in the rhizosphere led to the increase of the plant dry weight in the symbiosis between Peking and USDAI93, and that between Kegonnotaki and MAFF03-03063. On the other hand, dry matter accumulation in the symbiosis between Akisengoku and AI017 decreased under the supply of 25 dm³ m^-3 CO₂ air aimed at increasing the CO₂ concentration in the rhizosphere beyond the optimum CO₂ concentration for growth. Nodule mass and nodule number per plant were highest in Akisengoku, followed by Kegonnotaki and lowest in Peking. Also the increase of the CO₂ concentration in the rhizosphere led to the increase of the nodule mass and number in Kegonnotaki, while no changes were observed in Akisengoku and Peking. Biological nitrogen fixation (BNF) was highest in Akisengoku, followed by Kegonnotaki, and lowest or near zero in Peking. BNF in Akisengoku and Kegonnotaki showed a similar tendency to that of dry matter accumulation. BNF of Peking was especially low under the supply of CO₂-free air, and it increased with the increase of the CO₂ concentration in the rhizosphere. For the symbiosis of Bradyrhizobium strains with soybean and cowpea, the most suitable CO₂ concentration for N₂ fixation and plant growth was estimated to be about 10 dm³ m^-3, while for the symbiosis of S. fredii with soybean, the value was estimated to be above 30 dm³ m^-3.     

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。     

Real-time imaging of nitrogen fixation in an intact soybean plant with nodules using 13N-labeled nitrogen gas

Abstract

Real-time images of nitrogen fixation in an intact nodule of hydroponically cultured soybean (Glycine max [L] Merr.) were obtained. In the present study, we developed a rapid method to produce and purify ¹³N-labeled radioactive nitrogen gas (half life: 9.97?min). ¹³N was produced from a ¹⁶O (p, α) ¹³N nuclear reaction. The target chamber was filled with CO₂ and irradiated for 10?min with protons at an energy of 18.3?MeV and an electric current of 5?μA, which was delivered from a cyclotron. All CO₂ in the collected gas was absorbed and removed with powdered soda-lime in a syringe and replaced with helium gas. The resulting gas was injected into gas chromatography and separated and a 35?mL fraction, including the peak of [¹³N]-nitrogen gas, was collected by monitoring the chromatogram. The obtained gas was mixed with 10?mL of O₂ and 5?mL of N₂ and used in the tracer experiment. The tracer gas was fed into the underground part of intact nodulated soybean plants and serial images of the distribution of ¹³N were obtained non-invasively using a positron-emitting tracer imaging system (PETIS). The rates of nitrogen fixation of the six test plants were estimated to be 0.17?±?0.10?μmol N₂?h^?1 from the PETIS image data. The decreasing rates of assimilated nitrogen were also estimated to be 0.012?±?0.011?μmol?N₂?h^?1. In conclusion, we successfully observed nitrogen fixation in soybean plants with nodules non-invasively and quantitatively using [¹³N]N₂ and PETIS.     

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.     

Comparison of 15N natural abundance and difference methods for estimating N2 fixation of soybeans grown in a tropical soil

Abstract

A study was carried out to compare the difference or N-yield method with the ¹⁵N natural abundance method for the estimation of the fractional contribution of biological N₂ fixation in the different plant parts of nodulating and non-nodulating isolines of soybeans. The results indicated that the δ¹⁵N values of most plant parts of soybeans were significantly lower (p<0.05) in the nodulating than in the non-nodulating isoline. However, in the case of the root+nodule component, the δ¹⁵N value was higher in the nodulating than in the non-nodulating isoline possibly due to isotopic discrimination of ¹⁵N over ¹⁴N which may have occurred in the nodules. Inoculation of soybeans with the Bradyrhizobium japonicum strain CB 1809 increased significantly (p<0.05) the δ¹⁵N value of the root+nodule component implying that the effectiveness of the soybean-rhizobium symbiosis had increased by inoculation.

Percentage of plant N derived from atmospheric N₂ fixation (%Ndfa) estimated by the ¹⁵N natural abundance method was highly correlated (r=0.762, p<0.01) with that by the difference or N-yield method and the differences between the two methods were not statistically significant. The agreement between the two methods was closer at maturity than at the early reproductive stage.

The %Ndfa obtained by the difference method ranged from 48.4 to 92.6% whereas the %Ndfa obtained by the ¹⁵N natural abundance method ranged from 43.2 to 92.4% in the different plant parts. Based on the ¹⁵N natural abundance method, approximately 15% of the N in pod, shoot, grain, and shell was derived from the soil but in the case of stover, this fraction was about 55%.     

玉米/大豆、玉米/花生间作对作物氮素吸收及结瘤固氮的影响

禾本科与豆科作物间作具有显著的增氮作用。为探明玉米/大豆、玉米/花生间作模式的氮素吸收、氮营养竞争能力及豆科结瘤特性的变化,解释玉米与豆科间作体系的增氮效应,通过田间试验,设置玉米单作（MM）、大豆单作（SS）、玉米/大豆间作（MS）、花生单作（PP）、玉米/花生间作（MP）等5种种植模式,研究不同种植模式对作物氮素积累、氮营养竞争强弱及豆科结瘤固氮特性的调控作用。结果表明,与单作相比,间作显著降低玉米和大豆的氮素积累量,对花生的氮素积累量影响不显著。5种模式系统氮素积累总量表现为MS > SS > MP,PP和MM处理最低且差异不显著,MS处理比MP处理显著高21.8%。与MM处理相比,MS和MP处理的玉米氮素积累量分别降低20.5%和11.7%,其中MP处理籽粒、叶片和茎秆氮素积累量比MS处理高8.9%、21.2%和14.3%。与SS处理相比,MS处理的大豆氮素积累量降低28.5%,其中,中行、边行分别降低10.1%、15.4%。玉米相对大豆氮营养竞争比率表现为强（CR_ms>1）,相对花生则表现为弱（CR_mp<1）。与SS处理相比,五叶期MS处理的大豆根瘤数量显著增加,根瘤鲜重无显著差异,盛花期后根瘤数量和鲜重均显著降低;MS处理的大豆根瘤固氮酶活性均降低,且中行降低幅度更大。与PP处理相比,开花期MP处理的花生根瘤数量和鲜重均显著增加,下针期后均显著降低;MP处理的花生根瘤固氮酶活性均降低,且边行降低幅度更大。各间作模式作物的氮素积累量虽然降低,但间作模式的系统氮素积累量却显著高于各单作模式,两种间作模式中MS处理的氮素积累总量最高。     

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     

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.     

Forage nitrogen yield and soil nitrogen in artificial grasslands with varied Medicago seedling proportion

ABSTRACT

This study aimed to understand the effects of Medicago spp proportion on symbiotic and non-symbiotic nitrogen (N) utilization of plants, and subsequent forage production and soil N status in an artificial Leymus chinensis grassland. By a two-year field experiment conducted in semi-arid northern China, it was found that the corresponding biomass proportions of legume in swards were 0, 39, 63, 83 and 98% when legume seedling proportions at sowing were 0, 25, 50, 75 and 100%, respectively. Increased Medicago seedling proportion (from 25 to 100%) decreased legume N₂ fixation capacity from 53 to 21%, as a consequence, this reduced total symbiotic N₂ fixation and its contribution to forage production with Medicago seedling proportion increasing from 50 to 75% or more. However, as increased Medicago seedling proportion enhanced legume biomass and sward uptake to soil mineral N, higher legume stands still led to the greater biomass and N yield. The cultures with 50% seedling of legume had 4–13% greater soil N concentration than the cultures with 0, 25, 75 and 100% seedling of legume. We concluded that pure Medicago stands led to the greatest forage yield, while medium Medicago stands could lead to the greater symbiotic N fixation and soil N concentration.     

Atmospheric nitrogen fixation by photosynthetic microorganisms in a submerged Philippine soil

Photosynthetic nitrogen-fixing microorganisms help maintain the nitrogen level of soil in rice paddies when environmental factors favor the growth of microorganisms. Our studies showed that blue-green algae in particular have a significant role in nitrogen-fixation in light. The most active nitrogen-fixation by microorganisms occurred in the soil shortly after it had been submerged under light. The longer the submergence, the less nitrogen microorganisms were fixed. In a greenhouse experiment, the fixed nitrogen appeared not to be immediately available to the rice plant. The amount of nitrogen that can be fixed in the field by nitrogen-fixing microorganisms in paddy water was estimated using the acetylene reduction method during the rice-growing period. The amount of nitrogen fixation by these microorganisms is not sufficient to account for the amount of nitrogen uptake by rice during the rice-growing period.     

Effects of land-use type and nitrogen addition on nitrous oxide and carbon dioxide production potentials in Japanese Andosols

Land-use type and nitrogen (N) addition strongly affect nitrous oxide (N₂O) and carbon dioxide (CO₂) production, but the impacts of their interaction and the controlling factors remain unclear. The aim of this study was to evaluate the effect of both factors simultaneously on N₂O and CO₂ production and associated soil chemical and biological properties. Surface soils (0–10 cm) from three adjacent lands (apple orchard, grassland and deciduous forest) in central Japan were selected and incubated aerobically for 12 weeks with addition of 0, 30 or 150 kg N ha^–1 yr^–1. Land-use type had a significant (p < 0.001) impact on the cumulative N₂O and CO₂ production. Soils from the apple orchard had higher N₂O and CO₂ production potentials than those from the grassland and forest soils. Soil net N mineralization rate had a positive correlation with both soil N₂O and CO₂ production rates. Furthermore, the N₂O production rate was positively correlated with the CO₂ production rate. In the soils with no N addition, the dominant soil properties influencing N₂O production were found to be the ammonium-N content and the ratio of soil microbial biomass carbon to nitrogen (MBC/MBN), while those for CO₂ production were the content of nitrate-N and soluble organic carbon. N₂O production increased with the increase in added N doses for the three land-use types and depended on the status of the initial soil available N. The effect of N addition on CO₂ production varied with land use type; with the increase of N addition doses, it decreased for the apple orchard and forest soils but increased for the grassland soils. This difference might be due to the differences in microbial flora as indicated by the MBC/MBN ratio. Soil N mineralization was the major process controlling N₂O and CO₂ production in the examined soils under aerobic incubation conditions.     

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.