Nonnodulated Soybean Plants Research Articles

Nitrogen Metabolism in Non-Nodulated and Nodulated Soybean Plants Related to Ureide Synthesis

Soybean plants can fix atmospheric N2 in the root nodule, a symbiotic organ with rhizobia. The primary forms of N transported from N2 fixation are ureides, allantoate, and allantoin, supplemented with asparagine. The nitrate absorbed in the roots is transported to the shoots in the forms of NO3− and asparagine with a little portion of ureides. The concentrations of N-metabolites were analyzed by capillary electrophoresis after supplying various concentrations of urea, precursors of ureides, and allopurinol, an inhibitor of xanthine dehydrogenase, to investigate the ureide synthesis pathway in the roots. When the non-nodulated soybean plants were treated with 0–5 mM of urea, the concentrations of asparagine and glutamine in the xylem sap and the roots increased remarkably. In addition, allantoate concentration increased with the urea concentrations becoming higher. Allopurinol inhibited the accumulation of allantoate but did not affect the asparagine and glutamine accumulation in roots, stems, leaves, and xylem sap, supporting that allantoate is synthesized by purine degradation in roots the same as in the nodules. When ureide precursors were supplied to the nodulated soybean plants, the concentrations of asparagine and glutamine in the xylem sap and roots increased, suggesting that the ureide precursors were absorbed and assimilated to amides in the roots.

Are phloem-derived amino acids the origin of the elevated malate concentration in the xylem sap following mineral N starvation in soybean?

A substantial increase in malate in the xylem sap of soybean subjected to mineral N starvation originates mainly from aspartate, a prominent amino acid of the phloem. A substantial increase in xylem malate was found when non-nodulated soybean plants were transferred to a N-free medium. Nodulated plants growing in the absence of mineral N and, therefore, dependent on symbiotic N2 fixation also contained elevated concentrations of malate in the xylem sap. When either nitrate or ammonium was supplied, malate concentrations in the xylem sap were low, both for nodulated and non-nodulated plants. Evidence was obtained that the elevated malate concentration of the xylem was derived from amino acids supplied by the phloem. Aspartate was a prominent component of the phloem sap amino acids and, therefore, a potential source of malate. Supplying the roots of intact plants with 13C-aspartate revealed that malate of the xylem sap was readily labelled under N starvation. A hypothetical scheme is proposed whereby aspartate supplied by the phloem is metabolised in the roots and the products of this metabolism cycled back to the shoot. Under N starvation, aspartate metabolism is diverted from asparagine synthesis to supply N for the synthesis of other amino acids via transaminase activity. The by-product of aspartate transaminase activity, oxaloacetate, is transformed to malate and its export accounts for much of the elevated concentration of malate found in the xylem sap. This mechanism represents a new additional role for malate during mineral N starvation of soybean, beyond that of charge balance.

Provision of carbon skeletons for amide synthesis in non-nodulated soybean and pea roots in response to the source of nitrogen supply

Soluble amino acids in roots and primary amino acids, which were involved in primary ammonium assimilation, in the metabolites of 14C-glucose fed to roots for 3 h in the dark were analyzed in the roots of non-nodulated soybean and pea plants grown in ammonium, nitrate or nitrogen-free media for 1 day. Compared with the effect of nitrate, ammonium supply strongly affected the content and synthesis of the amino acids in the roots. In both soybean and pea roots, the supply of ammonium increased considerably the concentrations of the primary amino acids, and asparagine was the most predominant amide, followed by glutamine. In nitrate-supplied soybean roots, the concentrations of asparagine, aspartate and alanine increased, but the concentration of glutamine was low. In the roots of pea plants grown in nitrate media, asparagine was the predominant amino acid, although the composition of the primary amino acids was little affected by nitrate supply. The proportion of amino acids synthesized from 14C-glucose increased and asparagine rather than glutamine was predominantly synthesized in ammonium-supplied soybean and pea roots, whereas in nitrate-supplied roots asparagine was more actively synthesized than glutamine, although asparagine was not predominant. The ratio of C4 (asparagine + aspartate) to C5 (glutamine + glutamate) amino acids was twofold higher in ammonium-supplied and nitrate-supplied soybean roots than in roots receiving no nitrogen. In contrast, in pea roots, the C4/C5 ratio was twofold higher only in ammonium nutrition. The results obtained suggest that the roots of leguminous plants might possess an indigenous ability to provide a carbon skeleton for preferential synthesis of asparagine rather than glutamine with a high intensity of ammonium supply.

Nitrogen stress and the expression of asparagine synthetase in roots and nodules of soybean (Glycine max)

The difficulty of assaying asparagine synthetase (AS) (EC 6.3.5.4) activity in roots of soybean has been circumvented by measuring expression of the AS genes. Expression of three soybean asparagine synthetase (SAS) genes (SAS1, SAS2 and SAS3) was observed in roots of non-nodulated soybean plants cultivated on nitrate. Expression of these genes was reduced to very low levels within days after submitting the plants to a N-free medium. The subsequent return to a complete medium (containing nitrate) restored expression of all three AS genes. Roots of nodulated plants, where symbiotic nitrogen fixation was the exclusive source of N (no nitrate present), showed very weak expression of all three AS genes, but on transfer to a nitrate-containing medium, strong expression of these genes was observed within 24 h. In nodules, all three genes were expressed in the absence of nitrate. Under conditions that impair nitrogen fixation (nodules submerged in aerated hydroponics), only SAS1 expression was reduced. However, in the presence of nitrate, an inhibitor of N(2) fixation, SAS1 expression was maintained. High and low expressions of AS genes in the roots were associated with high and low ratios of Asn/Asp transported to the shoot through xylem. It is concluded that nitrate (or one of its assimilatory products) leads to the induction of AS in roots of soybean and that this underlies the variations found in xylem sap Asn/Asp ratios. Regulation of nodule AS expression is quite different from that of the root, but nodule SAS1, at least, appears to involve a product of N assimilation rather than nitrate itself.

N-stress alters aspartate and asparagine levels of xylem sap in soybean

The influence of N-stress on the transport of amino acids in the xylem sap was studied for nodulated and non-nodulated soybean plants. The transfer of non-nodulated plants cultivated with nitrate or of nodulated plants dependent exclusively on N 2 fixation to hydroponics with N-free nutrient solution (N-stress), led to a pronounced increase in aspartate and fall in asparagine (Asn) in the xylem sap. Such a change was also observed during the ‘N-hunger’ phase of the growth cycle of nodulated plants, characterized by the transient yellowing of leaves and coinciding with the initial stages of nodule development. In this case, aspartate increased to around 68% of the xylem amino acids while Asn, normally the most abundant amino acid, fell to 10%. The changes were reversible on recovery of plants from N-stress. In the case of nodulated plants, the fall in xylem Asn during N-stress and its subsequent rise during recovery closely followed Asn synthetase activity in the nodule, while aspartate produced an inverse relationship. Aspartate was prominent in the phloem, independent of the treatment. The possibility that the phenomenon is related to the metabolism of aspartate recycled through the root system is discussed.

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.

Dry matter and nitrogen accumulation are not affected by superoptimal concentration of ammonium in flowing solution culture with pH control.

While it is known that superoptimal concentrations of the nitrate (NO3-) ion in solution culture do not increase NO3- uptake or dry matter accumulation, the same is not known for the ammonium (NH4+) ion. An experiment was conducted utilizing flowing solution culture with pH control to investigate the influence of superoptimal NH4+ concentrations on dry matter, nitrogen (N), potassium (K), calcium (Ca), and magnesium (Mg) accumulation by nonnodulated soybean plants. Increasing the NH4+ concentration in solution from 1 to 10 mM did not affect dry matter or N accumulation. Accumulations of K, Ca, and Mg were slightly decreased with increased NH4+ concentration. The NH4+ uptake system, which is saturated at less than 1mM NH4+, is able to regulate uptake of NH4+ at concentrations as high as 10 mM.

Presence of acetone and acetoacetate decarboxylase in soybean moot nodules

Abstract Acetone was detected in the leaves, roots, and nodules of soybean [Glycine max (L.) Merr.] plants, but not in the stems. The concentration of acetone in the nodules was the highest among the three former organs. Although acetone was also detected in the leaves and the roots of non-nodulated soybean plants grown with nitrate, the concentration in each case was half of that in nodulated soybeans. A small amount of acetone evolved from detached nodules but not from nodulated roots. The activity of acetoacetate decarboxylase that catalyzes the conversion of acetoacetate to acetone was detected in the nodules, mainly in the bacteroids. These results suggest that acetone is produced by the action of acetoacetate decarboxylase in soybean nodules. The luctuations of the acetone content in the nodules during their development were not correlated with those of the acetylene reduction activity. The correlation between the changes in the acetylene reduction activity and those of the acetone content in detach...

Nitrogen and phosphorus nutritional interactions in a CO2enriched environment

Abstract Nonnodulated soybean plants (Glycine max. [L.] Merr. ‘Lee') were supplied with nutrient solutions containing growth limiting concentrations of N or P to examine effects on N‐ and P‐uptake efficiencies (mg nutrient accumulated/gdw root) and utilization efficiencies in dry matter production (gdw2/mg nutrient). Nutritional treatments were imposed in aerial environments containing either 350 or 700 μL/L atmospheric CO2 to determine whether the nutrient interactions were modified when growth rates were altered. Nutrient‐stress treatments decreased growth and N‐ and P‐uptake and utilization efficiencies at 27 days after transplanting (DAT) and seed yield at maturity (98 DAT). Atmospheric CO2 enrichment increased growth and N‐ and P‐utilization efficiencies at 27 DAT and seed yield in all nutritional treatments and did not affect N‐ and P‐uptake efficiencies at 27 DAT. Parameter responses to nutrient stress at 27 DAT were not altered by atmospheric CO2 enrichment and vice versa. Nutrient‐stress treatments lowered the relative seed yield response to atmospheric CO2 enrichment. Decreased total‐N uptake by P‐stressed plants was associated with both decreased root growth and N‐uptake efficiency of the roots. Nitrogen‐utilization efficiency was also decreased by P‐stress. This response was associated with decreased plant growth as total‐N uptake and plant growth were decreased to the same extent by P stress resulting in unaltered tissue N concentrations. In contrast, decreased total P‐uptake by N‐stressed plants was associated with a restriction in root growth as P‐uptake efficiency of the roots was unaltered. This response was coupled with an increased root‐to‐shoot dry weight ratio; thus shoot and whole‐plant growth were decreased to a much greater extent than total‐P uptake which resulted in elevated P concentrations in the tissue. Therefore, P‐utilization efficiency was markedly reduced by N stress.

Cyclic variations in nitrogen uptake rate in soybean plants: uptake during reproductive growth.

Net uptake of NO3- by non-nodulated soybean plants [Glycine max (L.) Merr. cv. Ransom] growing in flowing hydroponic culture was measured daily during a 63 d period of reproductive development between the first florally inductive photoperiod and [unknown word] seed growth. Removal of NO3- from a replenished solution containing 1.0 mol m-3 NO3- was determined by ion chromatography. Uptake of NO3- continued throughout reproductive development. The net uptake rate of NO3- cycled between maxima and minima with a periodicity of oscillation of 3 to 7 d during the floral stage and about 6 d during the fruiting stage. Coupled with increasing concentrations of carbon and C : N ratios in tissues, the oscillations in net uptake rates of NO3- are evidence that the demand for carbohydrate by reproductive organs is contingent on the availability of nitrogen in the shoot pool rather than that the demand for nitrogen follows the flux of carbohydrate into reproductive tissues.

Alterations in Leaf Carbohydrate Metabolism in Response to Nitrogen Stress

A series of experiments was conducted to characterize alterations in carbohydrate utilization in leaves of nitrogen stressed plants. Two-week-old, nonnodulated soybean plants (Glycine max [L.] Merrill, ;Ransom'), grown previously on complete nutrient solutions with 1.0 millimolar NO(3) (-), were transferred to solutions without a nitrogen source at the beginning of a dark period. Daily changes in starch and sucrose levels of leaves were monitored over the following 5 to 8 days in three experiments. Starch accumulation increased relative to controls throughout the leaf canopy during the initial two light periods after plant exposure to N-free solutions, but not after that time as photosynthesis declined. The additional increments of carbon incorporated into starch appeared to be quantitatively similar to the amounts of carbon diverted from amino acid synthesis in the same tissues. Since additional accumulated starch was not degraded in darkness, starch levels at the beginning of light periods also were elevated. In contrast to the starch effects, leaf sucrose concentration was markedly higher than controls at the beginning of the first light period after the N-limitation was imposed. In the days which followed, diurnal turnover patterns were similar to controls. In source leaves, the activity of sucrose-P synthase did not decrease until after day 3 of the N-limitation treatment, whereas the concentration of fructose-2,6-bisphosphate was decreased on day 2. Restricted growth of sink leaves was evident with N-limited plants within 2 days, having been preceeded by a sharp decline in levels of fructose-2,6 bisphosphate on the first day of treatment. The results suggest that changes in photosynthate partitioning in source leaves of N-stressed plants resulted largely from a stable but limited capacity for sucrose formation, and that decreased sucrose utilization in sink leaves contributed to the whole-plant diversion of carbohydrate from the shoot to the root.

Cyclic variations in nitrogen uptake rate of soybean plants: effects of external nitrate concentration.

Net uptake of NO3- by non-nodulated soybean plants [Glycine max (L.) Merr. cv. Ransom] growing in flowing hydroponic cultures containing 0.5, 1.0 and 10.0 mol m-3 NO3- was measured daily during a 24-d period of vegetative development to determine if amplitude of maximum and minimum rates of net NO3- uptake are responsive to external concentrations of NO3-. Removal of NO3- from the replenished solutions during each 24-h period was determined by ion chromatography. Neither dry matter accumulation nor the periodicity of oscillations in net uptake rate was altered by the external NO3- concentrations. The maxima of the oscillations in net uptake rate, however, increased nearly 3-fold in response to external NO3- concentrations. The maxima and minima, respectively, changed from 4.0 and 0.6 mmol NO3- per gram root dry weight per day at an external solution level of 0.5 mol m-3 NO3- to 15.2 and -2.7 mmol NO3- per gram root dry weight per day at an external solution level of 10.0 mol m-3 NO3-. The negative values for minimum net uptake rate from 10.0 mol m-3 NO3- solutions show that net efflux was occurring and indicate that the magnitude of the efflux component of net uptake was responsive to external concentration of NO3-.

Endogenous NO3− in the Root as a Source of Substrate for Reduction in the Light

An experiment was conducted to investigate the reduction of endogenous NO(3) (-), which had been taken up by plants in darkness, during the course of the subsequent light period. Vegetative, nonnodulated soybean plants (Glycine max [L]. Merrill, ;Ransom') were exposed to 1.0 millimolar (15)NO(3) (-) for 12 hours in darkness and then returned to a solution containing 1.0 millimolar (14)NO(3) (-) for the 12 hours ;chase' period in the light. Another set of plants was exposed to (15)NO(3) (-) during the light period to allow a direct comparison of contributions of substrate from the endogenous and exogenous sources. At the end of the (15)NO(3) (-) exposure in the dark, 70% of the absorbed (15)NO(3) (-) remained unreduced, and 83% of this unreduced NO(3) (-) was retained in roots. The pool of endogenous (15)NO(3) (-) in roots was depleted at a steady rate during the initial 9 hours of light and was utilized almost exclusively in the formation of insoluble reduced-N in leaves. Unlabeled endogenous NO(3) (-), which had accumulated in the root prior to the previous dark period, also was depleted in the light. When exogenous (15)NO(3) (-) was supplied during the light period, the rate of assimilation progressively increased, reflecting an increased rate of uptake and decreased accumulation of NO(3) (-) in the root tissue. The dark-absorbed endogenous NO(3) (-) in the root was the primary source of substrate for whole-plant NO(3) (-) reduction in the first 6 hours of the light period, and exogenous NO(3) (-) was the primary source of substrate thereafter. It is concluded that retention of NO(3) (-) in roots in darkness and its release in the following light period is an important whole-plant regulatory mechanism which serves to coordinate delivery of substrate with the maximal potential for NO(3) (-) assimilation in photosynthetic tissues.

Nitrogen and dry-matter partitioning in soybean plants during onset of and recovery from nitrogen stress.

The study tested the hypothesis that resupplying nitrogen after a period of nitrogen stress leads to restoration of the balance between root and shoot growth and normal functional activity. Nonnodulated soybean plants were grown hydroponically for 14 days with 1.0 mM NO3- in a complete nutrient solution. One set of plants was continued on the complete nutrient solution for 25 days; a second set was given 0.0 mM NO3- for 25 days; and the third set was given 0.0 mM NO3- for 10 days followed by transfer to the complete solution with 1.0 mM NO3- for 15 days. In continuously nitrogen-stressed plants, emergence and expansion of main-stem and branch leaves were severely inhibited as low nitrogen content limited further growth. This was followed by a shift in partitioning of dry matter from the leaves to the roots, resulting in an initial stimulation of root growth and a decreased shoot:root ratio. Reduced nitrogen also was redistributed from the leaves into the stem and roots. When nitrogen stress was relieved, leaf initiation and expansion were renewed. With the restoration of the balance between root and shoot function, the shoot:root ratio and distribution of reduced nitrogen within the plant organs returned to levels similar to those of nonstressed plants.

Utilization of ammonium as a nitrogen source: effects of ambient acidity on growth and nitrogen accumulation by soybean.

Dry matter accumulation of plants utilizing NH4+ as the sole nitrogen source generally is less than that of plants receiving NO3- unless acidity of the root-zone is controlled at a pH of about 6.0. To test the hypothesis that the reduction in growth is a consequence of nitrogen stress within the plant in response to effects of increased acidity during uptake of NH4+ by roots, nonnodulated soybean plants (Glycine max [L.] Merr. cv Ransom) were grown for 24 days in flowing nutrient culture containing 1.0 millimolar NH4+ as the nitrogen source. Acidities of the culture solutions were controlled at pH 6.1, 5.1, and 4.1 +/- 0.1 by automatic additions of 0.01 N H2SO4 or Ca(OH)2. Plants were sampled at intervals of 3 to 4 days for determination of dry matter and nitrogen accumulation. Rates of NH4+ uptake per gram root dry weight were calculated from these data. Net CO2 exchange rates per unit leaf area were measured on attached leaves by infrared gas analysis. When acidity of the culture solution was increased from pH 6.1 to 5.1, dry matter and nitrogen accumulation were reduced by about 40% within 14 days. Net CO2 exchange rates per unit leaf area, however, were not affected, and the decreased growth was associated with a reduction in rates of appearance and expansion of new leaves. The uptake rates of NH4+ per gram root were about 25% lower throughout the 24 days at pH 5.1 than at 6.1. A further increase in solution acidity from pH 5.1 to 4.1 resulted in cessation of net dry matter production and appearance of new leaves within 10 days. Net CO2 exchange rates per unit leaf area declined rapidly until all viable leaves had abscised by 18 days. Uptake rates of NH4+, which were initially about 50% lower at pH 4.1 than at 6.1 continued to decline with time of exposure until net uptake ceased at 10 days. Since these responses also are characteristic of the sequence of responses that occur during onset and progression of a nitrogen stress, they corroborate our hypothesis.

Cyclic Variations in Nitrogen Uptake Rate in Soybean Plants

Uptake of NO(3) (-) by nonnodulated soybean plants (Glycine max L. Merr. cv Ransom) growing in flowing hydroponic culture at 22 and 14 degrees C root temperatures was measured daily during a 31-day growth period. Ion chromatography was used to determine removal of NO(3) (-) from solution during each 24-hour period. At both root-zone temperatures, rate of NO(3) (-) uptake per plant oscillated with a periodicity of 3 to 5 days. The rate of NO(3) (-) uptake per plant was consistently lower at 14 degrees C than 22 degrees C. The lower rate of NO(3) (-) uptake at 14 degrees C during the initial 5 to 10 days was caused by reduced uptake rates per gram root dry weight, but with time uptake rates per gram root became equal at 14 and 22 degrees C. Thereafter, the continued reduction in rate of NO(3) (-) uptake per plant at 14 degrees C was attributable to slower root growth.

Assimilation of 15NO3− Taken Up by Plants in the Light and in the Dark

An experiment was conducted to determine the extent that NO(3) (-) taken up in the dark was assimilated and utilized differently by plants than NO(3) (-) taken up in the light. Vegetative, nonnodulated soybean plants (Glycine max L. Merrill, ;Ransom') were exposed to (15)NO(3) (-) throughout light (9 hours) or dark (15 hours) phases of the photoperiod and then returned to solutions containing (14)NO(3) (-), with plants sampled subsequently at each light/dark transition over 3 days. The rates of (15)NO(3) (-) absorption were nearly equal in the light and dark (8.42 and 7.93 micromoles per hour, respectively); however, the whole-plant rate of (15)NO(3) (-) reduction during the dark uptake period (2.58 micromoles per hour) was 46% of that in the light (5.63 micromoles per hour). The lower rate of reduction in the dark was associated with both substantial retention of absorbed (15)NO(3) (-) in roots and decreased efficiency of reduction of (15)NO(3) (-) in the shoot. The rate of incorporation of (15)N into the insoluble reduced-N fraction of roots in darkness (1.10 micromoles per hour) was somewhat greater than that in the light (0.92 micromoles per hour), despite the lower rate of whole-plant (15)NO(3) (-) reduction in darkness.A large portion of the (15)NO(3) (-) retained in the root in darkness was translocated and incorporated into insoluble reduced-N in the shoot in the following light period, at a rate which was similar to the rate of whole-plant reduction of (15)NO(3) (-) acquired during the light period. Taking into account reduction of NO(3) (-) from all endogenous pools, it was apparent that plant reduction in a given light period ( approximately 13.21 micromoles per hour) exceeded considerably the rate of acquisition of exogenous NO(3) (-) (8.42 micromoles per hour) during that period. The primary source of substrate for NO(3) (-) reduction in the dark was exogenous NO(3) (-) being concurrently absorbed. In general, these data support the view that a relatively small portion (<20%) of the whole-plant reduction of NO(3) (-) in the light occurred in the root system.

Alterations in internal partitioning of carbon in soybean plants in response to nitrogen stress.

Alterations in internal partitioning of carbon were evaluated in plants exposed to limited nitrogen supply. Vegetative, nonnodulated soybean plants (Glycine max (L.) Merrill, 'Ransom') were grown for 21 days with 1.0 mM NO3- and then exposed to solutions containing 1.0, 0.1, or 0.0 mM NO3- for a 25-day treatment period. In nitrogen-limited plants, there were decreases in emergence of new leaves and in the expansion rate and final area at full expansion of individual leaves. As indicated by alterations in accumulation of dry weight, a larger proportion of available carbon in the plant was partitioned to the roots with decreased availability of nitrogen. Partitioning of reduced nitrogen to the root also was increased and, in plants devoid of an external supply, considerable redistribution of reduced nitrogen from leaves to the root occurred. The general decrease in growth potential and sink strength for nutrients in leaves of nitrogen-limited plants suggested that factors other than simply availability of nitrogen likely were involved in the restriction of growth in the leaf canopy and the associated increase in carbon allocation to the roots.

Relative Content of NO3− and Reduced N in Xylem Exudate as an Indicator of Root Reduction of Concurrently Absorbed 15NO3−

It is unclear if the relative content of NO(3) (-) and reduced N in xylem exudate provides an accurate estimate of the percentage reduction of concurrently absorbed NO(3) (-) in the root. Experiments were conducted to determine whether NO(3) (-) and reduced N in xylem exudate of vegetative, nonnodulated soybean plants (Glycine max [L.] Merr., ;Ransom') originated from exogenous recently absorbed (15)NO(3) (-) or from endogenous (14)N pools. Plants either were decapitated and exposed to (15)NO(3) (-) solutions for 2 hours or were decapitated for the final 20 minutes of a 50-minute exposure to (15)NO(3) (-) in the dark and in the light. Considerable amounts of (14)NO(3) (-) and reduced (14)N were transported into the xylem, but almost all of the (15)N was present as (15)NO(3) (-). Dissimilar changes in transport of (14)NO(3) (-), reduced (14)N and (15)NO(3) (-) during the 2 hours of sap collection resulted in large variability over time in the percentage of total N in the exudate which was reduced N. Over a 20-minute period the rate of (15)N transport into the xylem of decapitated plants was only 21 to 36% of the (15)N delivered to the shoot of intact plants. Based on the proportion of total (15)N which was found as reduced (15)N in exudate and in intact plants in the dark, it was estimated that 5 to 17% of concurrently absorbed (15)NO(3) (-) was reduced in the root. This was much less than the 38 to 59% which would have been predicted from the relative content of total NO(3) (-) and total reduced N in the xylem exudate.

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 KNO(3) 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 KNO(3), beginning at 65 days, C(2)H(2) reduction (N(2) 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 N(2) fixation.