Effect of Rhizobium sp. BARIRGm901 inoculation on nodulation,nitrogen fixation and yield of soybean (Glycine max) genotypes in gray terrace soil

Soybean plants require high amounts of nitrogen, which are mainly obtained from biological nitrogen fixation. A field experiment was conducted by soybean (Glycine max) genotypes, growing two varieties (Shohag and BARI Soybean6) and two advanced lines (MTD10 and BGM02026) of soybean with or without Rhizobium sp. BARIRGm901 inoculation. Soybean plants of all genotypes inoculated with Rhizobium sp. BARIRGm901 produced greater nodule numbers, nodule weight, shoot and root biomass, and plant height than non-inoculated plants. Similarly, inoculated plants showed enhanced activity of nitrogenase (NA) enzyme, contributing to higher nitrogen fixation and assimilation, compared to non-inoculated soybean plants in both years. Plants inoculated with Rhizobium sp. BARIRGm901 also showed higher pod, stover, and seed yield than non-inoculated plants. Therefore, Rhizobium sp. BARIRGm901 established an effective symbiotic relationship with a range of soybean genotypes and thus increased the nodulation, growth, and yield of soybean grown in gray terrace soils in Bangladesh.     

Relationship between Ureide N and N(2) Fixation, Aboveground N Accumulation, Acetylene Reduction, and Nodule Mass in Greenhouse and Field Studies with Glycine max L. (Merr)

The relationship between ureide N and N₂ fixation was evaluated in greenhouse-grown soybean (Glycine max L. Merr.) and lima bean (Phaseolus lunatus L.) and in field studies with soybean. In the greenhouse, plant N accumulation from N₂ fixation in soybean and lima bean correlated with ureide N. In soybean, N₂ fixation, ureide N, acetylene reduction, and nodule mass were correlated when N₂ fixation was inhibited by applying KNO₃ solutions to the plants. The ureide-N concentrations of different plant tissues and of total plant ureide N varied according to the effectiveness of the strain of Bradyrhizobium japonicum used to inoculate plants. The ureide-N concentrations in the different plant tissues correlated with N₂ fixation. Ureide N determinations in field studies with soybean correlated with N₂ fixation, aboveground N accumulation, nodule weight, and acetylene reduction. N₂ fixation was estimated by ¹⁵N isotope dilution with nine and ten soybean genotypes in 1979 and 1980, respectively, at the V9, R2, and R5 growth stages. In 1981, we investigated the relationship between ureide N, aboveground N accumulation, acetylene reduction, and nodule mass using four soybean genotypes harvested at the V4, V6, R2, R4, R5, and R6 growth stages. Ureide N concentrations of young stem tissues or plants or aboveground ureide N content of the four soybean genotypes varied throughout growth correlating with acetylene reduction, nodule mass, and aboveground N accumulation. The ureide-N concentrations of young stem tissues or plants or aboveground ureide-N content in three soybean genotypes varied across inoculation treatments of 14 and 13 strains of Bradyrhizobium japonicum in 1981 and 1982, respectively, and correlated with nodule mass and acetylene reduction. In the greenhouse, results correlating nodule mass with N₂ fixation and ureide N across strains were variable. Acetylene reduction in soybean across host-strain combinations did not correlate with N₂ fixation and ureide N. N₂ fixation, ureide N, acetylene reduction, and nodule mass correlated across inoculation treatments with strains of Bradyrhizobium spp. varying in effectiveness on lima beans. Our data indicate that ureide-N determinations may be used as an additional method to acetylene reduction in studies of the physiology of N₂ fixation in soybean. Ureide-N measurements also may be useful to rank strains of B. japonicum for effectiveness of N₂ fixation.     

Physiological changes in soybean (Glycine max) Wuyin9 in response to N and P nutrition

Phosphorus deficiency is a very common problem in the acid soil of central China. Previous research has shown that starter N and N topdressing at the flowering stage (Rl) increased soybean (Glycine max) yield and N₂ fixation (Gan et al, 1997, 2000). However, there is little information available concerning soybean response to P‐fertiliser in soybean production in central China (Gan, 1999). A field experiment was conducted to investigate the response to P (0 kg P ha^?1, 22 kg P ha^?1, 44 kg P ha^?1 before sowing) and N fertiliser application (N1: 0 kg N ha^?1, N2: 25 kg N ha^?1 before sowing, N3: N2 + 50 kg N ha^?1 at the V2 stage and N4: N2 + 50 kg N ha^?1 at the R1 stage) on growth, yield and N₂ fixation of soybean. Both N and P fertiliser increased growth and seed yield of soybean (P < 0.01). Application of basal P fertiliser at 22 kg P ha^?1 or 44 kg P ha^?1 increased total N accumulation by 11% and 10% (P < 0.01) and seed yield by 12% and 13% (P < 0.01), respectively, compared to the zero P treatment. Although application of starter N at 25 kg N ha^?1 had no positive effect on seed yield at any P level (P > 0.05), an application of a topdressing of 50 kg N ha^?1 at the V2 or R1 stage increased total N accumulation by 11% and 14% (P < 0.01) and seed yield by 16% and 21% (P < 0.01), respectively, compared to the zero N treatment. Soybean plants were grown on sterilised Perlite in the greenhouse experiment to study the physiological response to different concentrations of phosphate (P1: 0 mM; P2: 0.05 mM; P3: 0.5 mM; P4:1.0 mN) and nitrate (N1: 0 mM with inoculation, N2: 20 mM with inoculation). The result confirmed that N and P nutrients both had positive effects on growth, nodulation and yield (P < 0.01). The relative importance of growth parameters that contributed to the larger biomass with N and P fertilisation was in decreasing order: (i) total leaf area, (ii) individual leaf area, (iii) shoot/root ratio, (iv) leaf area ratio and (v) specific leaf area. The yield increase at N and P supply was mainly associated with more seeds and a larger pod number per plant, which confirmed the result from the field experiment.     

Effect of oxygen availability on nitrogen fixation by two Lotus species under flooded conditions

The pasture legumes Lotus uliginosus (Schk.) and Lotus corniculatus (L.), known to differ in their tolerance to flooding, were inoculated with Rhizobium loti and flooded for 60 d while subjected to two levels of dissolved pO2: 0.241 and 0.094 mol ml^-1. L. uliginosus showed significantly greater growth (shoot and root) and N2 fixation under both pO2s, compared to L. corniculatus, although growth and N2 fixation by L. corniculatus was not affected by the low pO2. Surprisingly, in L. uliginosus., growth, nodulation and N2 fixation were all increased by low pO2 while nodulation of L. corniculatus where low pO2 plants showed a significant increase over that of the higher pO2 plants while L. uliginosus plants showed a decline. Root porosity of L. uliginosus doubled in the low pO2-treatment from a mean of 14.5% in high pO2 roots to 28.5%, whereas that of L. corniculatus was relatively unaffected by pO2, being 7% and 9% for high and low pO2 plants, respectively. The structure of nodules differed little between species and treatments, although nodules/nodulated roots from the L. uliginosus plants had particularly profuse lenticels and aerenchyma. However, L. corniculatus nodules, especially those grown in the lower pO2 showed signs of early senescence with vacuolation of infected cells and green coloration when cut open. Leghaemoglobin (Lb) concentrations in nodules from both species were unaffected by low pO2, although that of L. corniculatus nodules, regardless of pO2, was significantly greater than L. uliginosus. Concentrations of the intercellular glycoprotein recognized by the monoclonal antibody MAC265 were significantly reduced in nodules from the low pO2 treatment in both species. Immunogold labelling showed that the MAC265 antigen was localized primarily within intercellular spaces within nodule cortices from both Lotus species. A marked decrease in deposition of the MAC265 antigen within the cortices of L. uliginosus nodules grown in the lower pO2, is discussed in terms of the relative abilities of the two Lotus spp. to maintain an O2 supply to the N2-fixing bacteroids within submerged nodules.Keywords: Lotus uliginosus, Lotus corniculatus, N2 fixation, flooding, oxygen.      

Understanding the variability in soybean nitrogen fixation across agroecosystems

Legume-based cropping systems have the potential to internally regulate N cycling due to the suppressive effect of soil N availability on biological nitrogen fixation. We used a gradient of endogenous soil N levels resulting from different management legacies and soil textures to investigate the effects of soil organic matter dynamics and N availability on soybean (Glycine max) N₂ fixation. Soybean N₂ fixation was estimated on 13 grain farm fields in central New York State by the ¹⁵N natural abundance method using a non-nodulating soybean reference. A range of soil N fractions were measured to span the continuum from labile to more recalcitrant N pools. Soybean reliance on N₂ fixation ranged from 36% to 82% and total N₂ fixed in aboveground biomass ranged from 40 to 224 kg N ha^?1. Soil N pools were consistently inversely correlated with % N from fixation and the correlation was statistically significant for inorganic N and occluded particulate organic matter N. However, we also found that soil N uptake by N₂-fixing soybeans relative to the non-nodulating isoline increased as soil N decreased, suggesting that N₂ fixation increased soil N scavenging in low fertility fields. We found weak evidence for internal regulation of N₂ fixation, because the inhibitory effects of soil N availability were secondary to the environmental and site characteristics, such as soil texture and corresponding soil characteristics that vary with texture, which affected soybean biomass, total N₂ fixation, and net N balance.     

Nitrate Inhibition of Legume Nodule Growth and Activity : II. Short Term Studies with High Nitrate Supply

Soybean plants (Glycine max [L.] Merr) were grown in sand culture with 2 millimolar nitrate for 37 days and then supplied with 15 millimolar nitrate for 7 days. Control plants received 2 millimolar nitrate and 13 millimolar chloride and, after the 7-day treatment period, all plants were supplied with nil nitrate. The temporary treatment with high nitrate inhibited nitrogenase (acetylene reduction) activity by 80% whether or not Rhizobium japonicum bacteroids had nitrate reductase (NR) activity. The pattern of nitrite accumulation in nodules formed by NR⁺ rhizobia was inversely related to the decrease and recovery of nitrogenase activity. However, nitrite concentration in nodules formed by NR⁻ rhizobia appeared to be too low to explain the inhibition of nitrogenase. Carbohydrate composition was similar in control nodules and nodules receiving 15 millimolar nitrate suggesting that the inhibition of nitrogenase by nitrate was not related to the availability of carbohydrate.

Nodules on plants treated with 15 millimolar nitrate contained higher concentrations of amino N and, especially, ureide N than control nodules and, after withdrawal of nitrate, reduced N content of treated and control nodules returned to similar levels. The accumulation of N₂ fixation products in nodules in response to high nitrate treatment was observed with three R. japonicum strains, two NR⁺ and one NR⁻. The high nitrate treatment did not affect the allantoate/allantoin ratio or the proportion of amino N or ureide N in bacteroids (4%) and cytosol (96%).

     

Hydrogen sulfide and rhizobia synergistically regulate nitrogen (N) assimilation and remobilization during N deficiency-induced senescence in soybean

Hydrogen sulfide (H₂S) is emerging as an important signalling molecule that regulates plant growth and abiotic stress responses. However, the roles of H₂S in symbiotic nitrogen (N) assimilation and remobilization have not been characterized. Therefore, we examined how H₂S influences the soybean (Glycine max)/rhizobia interaction in terms of symbiotic N fixation and mobilization during N deficiency-induced senescence. H₂S enhanced biomass accumulation and delayed leaf senescence through effects on nodule numbers, leaf chlorophyll contents, leaf N resorption efficiency, and the N contents in different tissues. Moreover, grain numbers and yield were regulated by H₂S and rhizobia, together with N accumulation in the organs, and N use efficiency. The synergistic effects of H₂S and rhizobia were also demonstrated by effects on the enzyme activities, protein abundances, and gene expressions associated with N metabolism, and senescence-associated genes (SAGs) expression in soybeans grown under conditions of N deficiency. Taken together, these results show that H₂S and rhizobia accelerate N assimilation and remobilization by regulation of the expression of SAGs during N deficiency-induced senescence. Thus, H₂S enhances the vegetative and reproductive growth of soybean, presumably through interactions with rhizobia under conditions of N deficiency.     

Phosphorus deficiency does not enhance proton release by roots of soybean [Glycine max (L.) Murr.]

Little work has been done on root exudation in soybean under P deficiency. This study examined the effect of P supply on release of protons and carboxylates by roots of soybean (Glycine max Heinong 35), and to correlate the release with excess uptake of cations over anions. Plants were either reliant on N₂ fixation or supplied with nitrate and were grown in nutrient solution with 1–50 μM P for 7 weeks. Release of protons and carboxylates from roots, and concentrations of Ca, Mg, K, Na, P, S, Cl and N in plants were measured weekly from week 4. Unlike in many other species, P deficiency decreased proton release per unit root biomass in N₂-fixing plants and increased release of hydroxyl ions in nitrate-fed soybean. While P deficiency generally decreased uptake of K, Ca, Mg, S, Cl and P, it increased nitrate uptake per unit root biomass. Irrespective of P supply, amounts of protons released correlated well with excess uptake of cations over anions by the roots. Phosphorus deficiency increased release of carboxylates but the amounts released were small. The results suggest that soybean displays strategies of P acquisition through decreasing proton release which favors P mobilization in acid soils, and increasing root-to-shoot ratio and specific root length.     

The response and recovery of nitrogen fixation activity in soybean to water deficit at different reproductive developmental stages

Soybean (Glycine max [L.] Merr.) N₂ fixation is a primary plant mechanism responsible for meeting plant-N demand during seed development. Nitrogen fixation is recognized as a drought-sensitive mechanism; however, N₂ fixation response to water deficit and N₂ fixation recovery at different reproductive stages are not well documented. We tested the hypothesis that water deficit during late reproductive stages would inhibit N₂ fixation and lead to the breakdown of essential leaf proteins and an inability to recover N₂ fixation. Acetylene reduction activity (ARA) and N redistribution response to a 5-d drought period at flowering (R2), early seed fill (R5), and late seed fill (R6) were evaluated in one genotype (Hendricks, maturity group 0). Control plants maintained high rates of nodule activity until late seed fill. Plants drought stressed at R2 and R5 recovered ARA after rewatering and in some cases had higher nitrogenase activity than control plants during mid-seed fill. Recovery of ARA on plants stressed at R2 and R5 was associated with higher shoot N concentration than control plants at maturity. Drought stress at R6 reduced ARA, and the inability to recover ARA after stress alleviation at R6 resulted in decreased individual seed mass, which was likely caused by an acceleration of leaf N redistribution and a shorter seed-fill period. Results emphasized the importance of soybean N₂ fixation during late seed development on seed yield and that the ability to recover N₂ fixation following drought is dependent upon crop developmental stage.     

Differences in photosynthetic behaviour and leaf senescence of soybean (Glycine max [L.] Merrill) dependent on N2 fixation or nitrate supply

Biological N₂ fixation can fulfil the N demand of legumes but may cost as much as 14% of current photosynthate. This photosynthate (C) sink strength would result in loss of productivity if rates of photosynthesis did not increase to compensate for the costs. We measured rates of leaf photosynthesis, concentrations of N, ureides and protein in leaves of two soybean cultivars ( Glycine max [L.] Merrill) differing in potential shoot biomass production, either associated with Bradyrhizobium japonicum strains, or amended with nitrate. Our results show that the C costs of biological N₂ fixation can be compensated by increased photosynthesis. Nodulated plants shifted N metabolism towards ureide accumulation at the start of the reproductive stage, at which time leaf N concentration of nodulated plants was greater than that of N-fertilized plants. The C sink strength of N₂ fixation increased photosynthetic N use efficiency at the beginning of plant development. At later stages, although average protein concentrations were similar between the groups of plants, maximum leaf protein of nodulated plants occurred a few days later than in N-fertilized plants. The chlorophyll content of nodulated plants remained high until the pod-filling stage, whereas the chlorophyll content of N-fertilized plants started to decrease as early as the flowering stage. These results suggest that, due to higher C sink strength and efficient N₂ fixation, nodulated plants achieve higher rates of photosynthesis and have delayed leaf senescence.     

Source of the soybean N credit in maize production

Nitrogen response trials throughout the United States Corn Belt show that economic optimum rates of N fertilization are usually less for maize (Zea mays L.) following soybean (Glycine max L.) than for maize following maize; however, the cause of this rotation effect is not fully understood. The objective of this study was to investigate the source of the apparent N contribution from soybean to maize (soybean N credit) by comparing soil N mineralization rates in field plots of unfertilized maize that had either nodulated soybean, non-nodulated soybean, or maize as the previous crop. Crop yields, plant N accumulation, soil inorganic N, and net soil mineralization were measured. Both grain yield (6.3 vs. 2.8 Mg ha^–1) and above-ground N accumulation (97 vs. 71 kg ha^–1) were greatly increased when maize followed nodulated soybean compared with maize following maize. A partial benefit to yield and N accumulation was also observed for maize following non-nodulated soybean. Cumulative net soil N mineralization following nodulated soybean, non-nodulated soybean, and maize was 112, 92 and 79 kg N ha^–1, respectively. Net mineralization of soil N appeared to be influenced by both quality (C:N ratio) and quantity of residue from the previous crop. In addition to an increase in plant available N from mineralization, the amount of soil inorganic N (especially in soil 5 cm from the row) was greater following nodulated soybean than non-nodulated soybean or maize. Based on these data, the soybean N credit appears to result from a combination of a decrease in net soil mineralization in continuous maize production and an increase in residual soil N from symbiotic fixation.     

Production of reactive oxygen species and antioxidant enzymes of pea and soybean plants under hypoxia and high CO<Subscript>2</Subscript> concentration in medium

Generation of reactive oxygen species (ROS) and activities of antioxidant enzymes (catalase, peroxidase, ascorbate peroxidase) in pea (Pisum sativum L.) and soybean (Glycine max L.) under hypoxia (3–24 h) and high CO₂ concentration in medium were studied. In sensitive to hypoxia pea seedlings, hypoxia enhanced markedly production of superoxide anion-radical, hydroperoxides, and especially hydrogen peroxide. In more tolerant soybean plants, these changes were less pronounced. During first hours of hypoxia, activity of lipoxygenase in plant cells increased. This allows a suggestion that this enzyme is involved in the processes of hydroperoxide accumulation in plant tissues under oxygen deficit. In pea and soybean plants, a correlation between tolerance to hypoxia, the rate of ROS generation, and antioxidant enzyme activities was established. During the first hours of hypoxia, the catalase activity in soybean plants increased stronger than in sensitive to hypoxia pea plants. At longer exposure to hypoxia (24 h), peroxidases started to play the higher role in cell defense against hypoxia, but only in soybean plants. The medium with the higher CO₂ content induced higher changes in the processes of ROS accumulation and activities of lipoxygenase and antioxidant enzymes. This permits us to refer CO₂, accumulated as a product of respiration in the cells, to low-molecular signal molecules switching on plant adaptation to hypoxic stress.     

N(2) fixation by bacteria associated with maize roots at a low partial o(2) pressure

Nitrogen fixation by bacteria associated with roots of intact maize plants was measured by exposing the roots to N(2) at a partial O(2) pressure (pO(2)) of 2 or 10 kPa. The plants were grown in a mixture of Weswood soil and sand and then transferred to plastic cylinders containing an N-free plant nutrient solution. The solution was sparged continuously with a mixture of air and N(2) at a pO(2) of 2 or 10 kPa. Acetylene reduction was measured after the roots were exposed to the low pO(2) overnight. The air-N(2) atmosphere in the cylinders was then replaced with an O(2)-He atmosphere at the same pO(2), and the roots were exposed to 20 kPa of N(2) for 20 to 22 h. Incorporation of N into the roots was 200 times greater at 2 kPa of O(2) than at 10 kPa of O(2). Adding l-malate (1 g of C liter) to the nutrient solution increased root-associated nitrogenase activity, producing a strong N label which could be traced into the shoots. Fixed N was detected in the shoots within 5 days after the plants were returned to unfertilized soil. In a similar experiment with undisturbed plants grown in fritted clay, movement of fixed N into the shoots was evident within 4 days after the roots were exposed to N(2) at 2 kPa of O(2). Inoculation with Azospirillum lipoferum yielded no significant differences in shoot dry weight, total nitrogen content, percent nitrogen, or N enrichment of plant tissues. Inoculated plants did exhibit greater root dry weight than uninoculated plants, however.     

Evaluation of the Relative Ureide Content of Xylem Sap as an Indicator of N(2) Fixation in Soybeans: GREENHOUSE STUDIES

The use of the relative ureide content of xylem sap [(ureide-N/total N) × 100] as an indicator of N₂ fixation in soybeans (Merr.) was examined under greenhouse conditions. Acetylene treatments to inhibit N₂ fixation were imposed upon the root systems of plants totally dependent upon N₂ fixation as their source of N and of plants dependent upon both N₂ fixation and uptake of exogenous nitrate. Significant decreases in the total N concentration of xylem sap from plants of the former type were observed, but no significant decrease was observed in the total N concentration of sap from the latter type of plants. In both types of plants, acetylene treatment caused significant decreases in the relative ureide content of xylem sap. The results provided further support for a link between the presence of ureides in the xylem and the occurrence of N₂ fixation in soybeans. The relative ureide content of xylem sap from plants totally dependent upon N₂ fixation was shown to be insensitive to changes in the exudation rate and total N concentration of xylem sap brought about by diurnal changes in environmental factors. There was little evidence of soybean cultivars or nodulating strains affecting the relative ureide content of xylem sap. `Ransom' soybeans nodulated with Rhizobium japonicum strain USDA 110 were grown under conditions to obtain plants exhibiting a wide range of dependency upon N₂ fixation. The relative ureide content of xylem sap was shown to indicate reliably the N₂ fixation of these plants during vegetative growth using a ¹⁵N method to measure N₂ fixation activity. The use of the relative ureide content of xylem sap for quantification of N₂ fixation in soybeans should be evaluated further.     

Understandings and future challenges in soybean functional genomics and molecular breeding

Soybean(Glycine max) is a major source of plant protein and oil.Soybean breeding has benefited from advances in functional genomics.In particular,the release of soybean reference genomes has advanced our understanding of soybean adaptation to soil nutrient deficiencies,the molecular mechanism of symbiotic nitrogen(N) fixation,biotic and abiotic stress tolerance,and the roles of flowering time in regional adaptation,plant architecture,and seed yield and quality.Nevertheless,many challenges remain...     

Effect of nodulation on the nitrate assimilation in vegetative soybean plants

Summary Nitrate assimilation in the first trifoliate leaf of vegetative soybean plants (Glycine max L. Merr, cv Hodgson) was studied in relation to nodulation. Nodulated and non-nodulated plants were grown in a nitrate medium (4 mM). As a control nodulated plants were grown in a nutrient medium without combined nitrogen. This study included measurements of the acetylene reduction activity of the whole plant and of thein vitro nitrate reductase, glutamine synthetase and glutamate dehydrogenase activities in the first leaf and of the nitrate concentration. Nitrate accumulation and nitrate reductase activity were depressed in nodulated plants; root growth was decreased in the presence of nitrate. The relationships between nitrate assimilation and nodulation are discussed.     

Inhibition of Nitrogen Fixation in Soybean Plants Supplied with Nitrate I. Nitrite Accumulation and Formation of Nitrosylleghemoglobin in Nodules

The accumulation of nitrite in nodules was investigated to elucidatethe mechanism of inhibition of nitrogen fixation in nodulesof soybean (Glycine max. [L.] Merr.) plants supplied with nitrate.Acetylene-reducing activity (ARA) in nodules fell within 24h as a result of the supply of exogenous nitrate, accompaniedby an increase in the accumulation of nitrite in the cytosolbut not in the bacteroids of nodules. Nitrate reductase (NR)activity in the nodule cytosol remained high, irrespective ofthe supply of nitrate. Nitrosylleghemoglobin (LbNO) was detectedspectrophotometrically in the extract from nodules in whichnitrogen fixation was inhibited by nitrate. In experiments invitro, it was found that LbNO was easily formed from leghemoglobinin the presence of nitrite and dithionite. Thus, it is suggested that nitrogen fixation was inhibited primarilyby a decrease in the function of leghemoglobin, attributableto the formation of LbNO, which was caused by the accumulationof nitrite generated from nitrate by NR in the nodule cytosol. (Received August 22, 1989; Accepted January 24, 1990)     

Growth,nutrient dynamics,and efficiency responses to carbon dioxide and phosphorus nutrition in soybean

Plant mineral nutrients such as phosphorus may exert major control on crop responses to the rising atmospheric carbon dioxide (CO₂) concentrations. To evaluate the growth, nutrient dynamics, and efficiency responses to CO₂ and phosphorus nutrition, soybean (Glycine max (L.) Merr.) was grown in controlled environment growth chambers with sufficient (0.50 mM) and deficient (0.10 and 0.01 mM) phosphate (Pi) supply under ambient and elevated CO₂ (aCO₂, 400 and eCO₂, 800 µmol mol^?1, respectively). The CO₂ × Pi interaction was detected for leaf area, leaf and stem dry weight, and total plant biomass. The severe decrease in plant biomass in Pi-deficient plants (10–76%) was associated with reduced leaf area and photosynthesis (P_net). The degree of growth stimulation (0–55% total biomass) by eCO₂ was dependent upon the severity of Pi deficiency and was closely associated with the increased phosphorus utilization efficiency. With the exception of leaf and root biomass, Pi deficiency decreased the biomass partitioning to other plant organs with the maximum decrease observed in seed weight (8–42%) across CO₂ levels. The increased tissue nitrogen (N) concentration in Pi-deficient plants was accredited to the lower biomass and increased nutrient uptake due to the larger root to shoot ratio. The tissue P and N concentration tended to be lower at eCO₂ versus aCO₂ and did not appear to be the main cause of the lack of CO₂ response of growth and P_net under severe Pi deficiency. The leaf N/P ratio of >16 was detrimental to soybean growth. The tissue P concentration needed to attain the maximum productivity for biomass and seed yield tended to be higher at eCO₂ versus aCO₂. Therefore, the eCO₂ is likely to increase the leaf critical P concentration for maximum biomass productivity and yield in soybean.     

Transport of nitrogen in the xylem of soybean plants

Experiments were conducted to characterize the distribution of N compounds in the xylem sap of nodulated and nonnodulated soybean plants through development and to determine the effects of exogenous N on the distribution of N compounds in the xylem. Xylem sap was collected from nodulated and nonnodulated greenhouse-grown soybean plants (Glycine max [L.] Merr. “Ransom”) from the vegetative phase to the pod-filling phase. The sum of the nitrogen in the amino acid, nitrate, ureide (allantoic acid and allantoin), and ammonium fractions of the sap from both types of plants agreed closely with total N as assayed by a Kjeldahl technique. Sap from nodulated plants supplied with N-free nutrient solution contained seasonal averages of 78 and 20% of the total N as ureide-N and amino acid-N, respectively. Sap from nonnodulated plants supplied with a 20 millimolar KNO₃ nutrient solution contained seasonal averages of 6, 36, and 58% of total N as ureide-N, amino acid-N, and nitrate-N, respectively. Allantoic acid was the predominant ureide in the xylem sap and asparagine was the predominant amino acid. When well nodulated plants were supplied with 20 millimolar KNO₃, beginning at 65 days, C₂H₂ reduction (N₂ fixation) decreased relative to nontreated plants and there was a concomitant decrease in the ureide content of the sap. A positive correlation (r = 0.89) was found between the ureide levels in xylem sap and nodule dry weights when either exogenous nitrate-N or urea-N was supplied at 10 and 20 millimolar concentrations to inoculated plants. The results demonstrate that ureides play a dominant role in N transport in nodulated soybeans and that the synthesis of ureides is largely dependent upon nodulation and N₂ fixation.