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.     

Growth and Nitrogen Fixation in Soybean as Affected by Phosphorus Fertilizer and Sheep Manure using 15N Isotopic Dilution

A field experiment was conducted to study the effect of adding different phosphorus (P) fertilizer levels [0, 40, and 80 kg phosphorus pentoxide (P₂O₅) ha^?1 (abbreviated as P0, P1, and P2, respectively)] and rates of sheep manure (M) [0, 20, and 40 ton ha^?1 (abbreviated as M0, M1, and M2, respectively)] on growth and nitrogen (N₂) fixation of soybean (Glycine max L.). Sorghum bicolor L. was employed as a reference crop to evaluate N₂ fixation using the ¹⁵N-isotpic dilution technique. Results showed that addition of P fertilizer or sheep manure had positive effects on dry-matter production, N accumulation, and seed yield. Such effects were more pronounced when adding sheep manure and P together than adding separately. Solely P fertilizer had a small impact on N₂ fixation. A tangible increase in the amounts of N₂ fixed due to manure addition occurred. The efficient use of N fertilizer (%NUE) increased significantly as the result of adding a high level of P fertilizer. However, a drastic decrease in %NUE was observed when sheep manure was added solely or in combination with P fertilizer. From productivity and ecological standpoints, P2M1 and P2M2 surpassed the other treatments in showing greater grain yield and greater N₂ fixation. However, considering the high cost of sheep manure, P2M1 was the optimal treatment for improving growth and N₂ fixation in soybean plants with minimal manure consumption. In conclusion, the integrated use of manure and P fertilizer could be considered a useful agricultural practice for improving the performance of soybean plants grown in an Aridisol. Their beneficial effects were mainly attributed to the enhancement of N₂ fixation through root growth and soil property improvements besides being a source of P and other nutrients that are essential for N₂-fixation process.     

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.     

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 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.     

不同利用方式红壤反硝化势和气态产物排放特征

采用厌氧培养-乙炔抑制法测定了4种不同利用方式红壤的反硝化势和气态产物N₂O和N₂的排放速率。结果表明,不同利用方式红壤反硝化势和N₂O和N₂的排放速率差异明显,土壤反硝化势强弱顺序依次为：竹林>茶园>林地>旱地。反硝化势与土壤有机碳(P<0.05)、厌氧培养期间土壤CO₂累积排放量(P<0.01)、nirS基因丰度( P<0.05)和nirK基因丰度(P<0.05) 呈显著正相关关系。逐步回归分析结果表明,CO₂累积排放量表征的易矿化碳是造成不同利用方式红壤反硝化势差异的主要原因,可以解释反硝化势变化的66%(P<0.01)。不同利用方式红壤N₂O和N₂排放速率差异明显,旱地红壤N₂O和N₂排放速率均最低,表明土壤pH的提升并没有增加旱地红壤的反硝化损失风险和N₂O排放速率。土壤易矿化有机碳含量也是影响不同利用方式红壤N₂O和N₂排放速率的主要因素。反硝化功能基因nirS、nirK和nosZ的丰度均与CO₂累积排放量呈显著正相关关系,进一步支持了土壤易矿化有机碳含量是影响不同利用方式红壤反硝化势和气态产物排放的主要因子。土壤pH是影响不同利用方式红壤反硝化气态产物N₂/N₂O的主要因素,但是pH影响红壤N₂/N₂O的微生物机制仍需要进一步研究。     

Phosphorus Deficiency Delays the Onset of Nodule Function in Soybean

Effects of phosphorus (P) deficiency on nodulation were examined in soybean grown in nutrient solution for 7 weeks. Increasing P supply increased shoot growth of nitrogen (N₂)-fixing plants from week 5 and that of nitrate-fed plant from week 4 after treatment. Nitrogen (N₂)-fixing plants had a greater P requirement for maximum growth at week 5. Increasing P supply from 1 to 16 μ M increased N concentration in N₂-fixing plants at week 4 but did not affect it from week 5. By contrast, P deficiency increased N concentration in nitrate-fed plants. Increasing P supply improved nodule formation from week 3. Nodule mass was affected more by P supply than nodule number, which, in turn, was affected more than plant growth. However, P supply did not decrease nodule specific N₂ fixation from week 5. The results suggest that P deficiency impaired symbiotic N₂ fixation through delaying onset of nodule function and decreasing nodule development.     

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.     

15N标记的土壤估测某些豆科植物N2固定的温室实验

Pot experiments were carried out to estimate N2 fixation by vetch,milk vetch,sickle alfalfa and broadbean in pure stand using a ^15N-labelled soil.Winter wheat was used as the non-fixing control.The 15N-labelled soil used was prepared by growing corn-wheat-corn successively on a nearly organic-matter-free Xiashu loess supplemented with adequate amounts of (15NH4)2SO4,P,K and micronutrients,then incorporating these 15N-labelled plant materials into the soil after each havest,and allowing the plant materials to be decomposed aerobically for 410d after incorporation of the plant material of the thire crop.The 15N enrichment of wheat plant-N varied slightly with organs,with a maximum difference of 9.8%,Based on 15N enrichment of soil N inferred from the mean value of the 15N enrichment in different organs of wheat 79%-91% of total N in the tops and 67%-74% of total N in the roots of legumes studied were derived from atmosphere .Estimate by isotope dilution method was in good agreement with that by the conventional difference method provided values obtained by the latter were corrected for seed N,and also with that from the measurement of N accumulated in the tops of the legumes.     

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.     

Nitrogen requirements associated with improved conservation tillage for corn production

Abstract

Excessive soil erosion and use of nitrogen fertilizer are costly to the Atlantic Coastal Plain corn (zea mays L., ’Funks G 4507') producer and both may serve to create environmental hazards. An in‐row chisel (36 cm deep) tillage method was compared with the standard 5 cm fluted coulter method for planting corn in premature wheat (Triticim aestivum L.) residues grown on an Orangeburg sandy loam (Typic Paleudult). Five orthogonal N levels ranging up to 440 kg of N/ha were used to determine an economic N optimum for each tillage method.

The in‐row chisel tillage method provides a possible yield advantage in the Atlantic Coastal Plain because of observed restricting soil layers within the normal corn rooting zone. The estimated profit‐maximizing quantities of N fertilizer were 262 and 295 kg of N/ha (234 and 263 1b of N/acre) for the fluted coulter and in‐row tillage procedures, respectively. Corn grain yields associated with these inputs were 9.6 × 10³ and 12.6 × 10³ kg/ha (153 and 200 bu/acre), respectively. The yield increase associated with in‐row chiseling through a 2.5 metric ton mulched surface is attributed to potentially improved rooting and more efficient water storage and use.