Nitrogen Mobilization from Stubble and Roots during Re-growth of Defoliated Perennial Ryegrass

Nitrogen re-mobilization during ryegrass (Lolium perenne L.)re-growth was studied as a function of time. Roots and stubblecompounds were labelled with ¹⁵N before clipping. Experimentalresults clearly showed that ryegrass re-growth involves twoperiods of differing physiological significance. The first occursduring the first 6 d when nearly all the nitrogen of the newleaves comes from organic nitrogen re-mobilized from roots andstubble. At the same time, the uptake of nitrogen from the mediumis very low, no reduction of endogenous nitrate occurs and proteolysisand amino amido nitrogen re-mobilization in stubble are notfully compensated for by synthesis.Thus, the change in protein-Ncontent, expressed per plant, in stubble decreases from 998µg (uncut plant) to 866 µg and 661 µg for2 d and 4 d respectively after cutting. The second period (afterthe sixth day) may be more representative of stable behaviourof ryegrass, with assimilation of mineral nitrogen from themedium providing the predominant source of nitrogen for re-growth. Key words: Lolium perenne L., nitrogen, re-growth     

Phloem Loading and Metabolism of Xylem-Borne Amino Compounds in Fruiting Shoots of a Legume

Cut, fruiting shoots of Lupinus albus L. supplied with ¹⁴C-and ¹⁵N-labelled L-asparagine, L-glutamine, L-aspartic acid,or L-glutamic acid through the transpiration stream readilytransferred the labelled carbon and nitrogen of each compoundto pods and seeds of fruits. A time course of labelling of phloemsap collected from petioles and fruit tips following feedingof labelled asparagine indicated that xylem to phloem exchangein leaflets was an immediate and effective route of transferof the amide to fruits and that this and the loading onto phloemof additional asparagine from unlabelled pools of the amidein stems furnished a major source of the nitrogen for fruitfilling. Xylem to phloem exchange of nitrogen was accomplishedin different ways for each amino acid. The amide nitrogen ofasparagine was transferred mainly in the unmetabolized compound,the nitrogen of aspartate and glutamate largely in a wide rangeof amino acids synthesized in the leaf, and the amide nitrogenof glutamine was transferred in a manner intermediate betweenthese extremes. Glutamine and asparagine were the principalphloem solutes labelled with nitrogen from any of the suppliedcompounds, but the photosynthetically produced amino acids,glutamate, aspartate, serine, alanine, and valine also became¹⁵N-labelled in phloem. The main pathway for glutamine synthesisin vegetative parts of the shoot appeared to be by amidationof glutamate, but asparagine was not considered to be derivedsimilarly from aspartate. Leaflets metabolized glutamine morereadily than asparagine, but in each case the amide nitrogenwas used for synthesis of a variety of amino acids and the carbonwas recovered largely in non-amino compounds.     

Changes in the amide and amino acid composition of xylem exudate from perennial ryegrass (Lolium perenne L.) during regrowth after defoliation

The paper presents the results of amino acid analyses in xylem sap during leaf regrowth of ryegrass plants defoliated firstly at the 8th and secondly at the 12th week of culture. The free amino acid composition of leaves, stubble and roots was also determined and some of the results are reported. Prior to defoliation, xylem sap contained a high proportion of amides, particularly glutamine. During regrowth after defoliation, the proportion of asparagine in the xylem sap increased until the third day when the highest ratios of asparagine/glutamine appeared. The results are compared with relative amounts of free amino acids in the different plant parts and discussed in relation to source-sink nitrogen transfer.     

Long-term Partitioning, Storage and Re-mobilization of 14C Assimilated by Lolium perenne (cv. Melle)

The youngest fully expanded leaves of single tillers of vegetativeperennial ryegrass plants were exposed to ¹⁴CO₂. Thereafter,quantitative and fractional analysis of the partitioning, storageand re-mobilization after defoliation of the ¹⁴C-labelled assimilatewas sequentially conducted over a 22 d period. In undefoliated plants, most ¹⁴C reached its final destinationwithin 5–6 of feeding. Forty per cent of assimilated ¹⁴Cwas subsequently lost through respiration, while 13.5, 8.5 and34 per cent remained in roots, stem bases and tops respectively.At least some ¹⁴C was distributed to tillers throughout theplant, but secondary tillers subtended by the fed tiller madethe greatest demand on ¹⁴C translocated from the fed tiller. A small, but significant portion of ¹⁴C was invested into longterm storage in undefoliated plants, four per cent of the totalassimilated still being present in a labile chemical form inroots and stem bases 22 d after feeding. In plants that wereseverely defoliated 4 d after feeding, depletion of reserve¹⁴C was observed relative to undefoliated plants. The depletiontook place from stem bases, not roots, and both low and highmolecular weight storage compounds were involved. A portionof the depleted ¹⁴C was incorporated into new growth after defoliation. Lolium perenne, perennial ryegrass, assimilate partitioning, storage, re-mobilization, defoliation     

Seasonal Variation of Nitrogenous Compounds in Components of the Kiwifruit Vine

Free amino acids in 6-year-old kiwifruit vines [Actinidia deliciosa(A. Chev.) C. F. Liang et A. R. Ferguson] were measured overthe course of 1 year using components obtained from whole-vineharvests. Tissues examined from the perennial structure consistedof the wood and cortex of structural roots, wood and bark ofstem, leader and 1-year-old fruiting canes. Free acids in theannual growth (fine roots, flowers, fruit, leaves and non-fruitingshoots) were also measured. The range of amino acids extracted indicated that kiwifruitconforms to a conventional pattern of nitrogen metabolism. Acidspresent in greatest concentration depended on tissue type andsampling time. In perennial components and fine roots, arginineand glutamine were the predominant species, followed by gamma-aminobutyrate,aspartate, glutamate, alanine and valine. Generally, maximumconcentrations of all free acids were measured in a 10-weekperiod around budbreak. These same acids, plus asparagine, serineand threonine, were also prevalent in annual growth. In leaves,flowers and non-fruiting shoots, concentrations were greatestin the young tissue and declined with age. By contrast, concentrationsof arginine, asparagine and glutamine in fruit peaked approximately10 weeks after anthesis, subsequent to the cell division phaseof growth. During the year, free arginine averaged 44, 48 and 58 % of thetotal N in the fine roots, and the cortex and wood of structuralroots, respectively (the quantity of total N and amino-N inother components of the structural framework was much less thanthat in root tissue). Arginine was the principal N-containingspecies measured in xylem sap vacuum-extracted from 1-year-oldcanes over winter. During the period of vegetative growth, however,glutamine and nitrate were the principal N-transport forms present.The study highlights the importance of the fine root systemas the primary location of nitrogenous reserves in this plantand identifies arginine as the dominant N-storage form. Actinidia deliciosa (A. Chev.) C. F. Liang et A. R. Ferguson, amino acid composition, kiwifruit, nitrogen, whole-plant harvesting     

Composition and Distribution of Free Amino Acids in Flatpea (Lathyrus sylvestris L.) as Influenced by Water Deficit and Plant Age

Drought-stressed flatpea (Lathyrus sylvestris L.) plants from8 to 22 weeks old were analysed for nitrogen, soluble proteinand free amino acids. An increase in nitrogen and free aminoacid concentrations and a decrease in soluble protein levelwere observed in roots of plants up to 16 weeks old. The cumulativeconcentration of free amino acids increased with drought stress.Tissue concentrations of 2, 4-diaminobutyric acid (1.6–2.6%of the dry weight) were highest in leaves. Levels increasedsteadily, nearly doubling, in leaves and stems between weeks10 and 16. Levels in drought-stressed leaves were, on average,11.9% higher than those of controls. Estimated concentrationsof a mixture of 4-aminobutyric acid and an unknown amino acidwere highest in stems, increased in this tissue with age andtended to increase in stems and leaves and decrease in rootsin response to water deficit. Levels of the mixture of homoserineand another unidentified amino acid were not influenced by ageor water status of the plants. Root concentrations of asparagine,arginine, glutamine, aspartate, and another prominent, unidentifiedamino acid increased with plant age and reached a peak at thetime of flowering (14 to 18 weeks). Only the concentration ofthe unknown compound was elevated following drought stress.Concentrations of valine, isoleucine, leucine, phenylalanine,and methionine also increased during this period and were elevatedin drought-stressed plants. Proline levels increased with plantage and drought stress, but proline accounted for only about10% of the total free amino acids in the drought-stressed plants. Key words: 2, 4-Diaminobutyric acid, drought, flatpea     

Seasonal Changes in Nitrogen Fractions of Coptis japonica, a Temperate Forest Evergreen Chamaephyte, and their Ecological Implications

Seasonal changes in several forms of nitrogen were investigatedin Coptis japonica, an evergreen rosette hemicryptophyte intemperate deciduous forest. The concentration of total nitrogenin rhizomes and roots decreased during the period of new shootgrowth from winter to spring. In the rhizomes, total solubleprotein stored by early summer decreased gradually until winter,coupled with an increase in free amino acids. Nitrogen was largelystored in free amino acids in the roots, especially during summer.The total soluble protein in current-year leaves decreased fromspring to summer and then increased during winter. The seasonalchanges in nitrogen components were coincident with the changein light-saturated photosynthetic rates recorded in a previousstudy. The ratio of total soluble protein to total nitrogendecreased from spring to summer and then increased from latesummer to winter in the current-year leaves. In contrast, chlorophyllcontent and the ratio of chlorophyll to total nitrogen werehigher in summer than in other seasons. The results indicatethat nitrogen was used in a manner that better utilizes thevery weak light in summer and the higher light intensities inother seasons. The major component of the free amino acid poolwas asparagine, in every organ throughout the season, exceptfor the senescent leaves. Since asparagine has a high N:C ratio,we suppose that the asparagine-dominated amino acid pool isadvantageous in the carbon-limited environment of the forestfloor.Copyright 1994, 1999 Academic Press Free amino acid composition, total nitrogen, total soluble protein, photosynthesis, evergreen hemicryptophyte     

Seasonal Changes of Nitrogen Storage Compounds in a Rhizomatous Grass Calamagrostis epigeios

The seasonal dynamics in content and distribution of N-rich compounds between overwintering organs of Calamagrostis epigeios were examined. Samples were taken both from plants grown in natural conditions and in containers with controlled nutrient supply. There were significant changes in content of nitrate, free amino acids and soluble protein in all investigated plant parts during the course of a year. Amino acids showed both the highest maximum and seasonal fluctuation among the all N compounds observed and, therefore, appear to have a central role in N storage. Their content rises in the autumn, remains stable during winter and declines quickly at the beginning of spring. The most abundant amino acids in the end of winter storage period - asparagine, arginine and glutamine - constituted about 90 % of N in fraction of free amino acids. The portion of N stored in soluble proteins, however, was considerably smaller compare to both amino acids and nitrate. The amount of N stored in rhizomes of C. epigeios was smaller than in roots and stubble base before the onset of spring re-growth. This indicates that roots and stubble base are particularly important for winter N storage in this species.     

Origin of Amino Acids in Datura stramonium Seeds

¹⁴C isotope studies show that the seeds of Datura stramoniumL. can produce a number of amino acids (particularly alanine,glutamate, phenylalanine, and tyrosine) from a supply of sucroseand nitrate. These amino acids can be incorporated into theseed protein. The bulk of the amino acids incorporated into the seed proteinmust, however, be supplied by adult leaves in the proximityof the fruit, either as the amino acids themselves, or theirimmediate precursors. The major free-amino-acid products of Datura leaves are theamides asparagine and glutamine.     

Increased Defoliation Frequency Depletes Remobilization of Nitrogen for Leaf Growth in Grasses

Plants ofLolium perenneandFestuca rubrawere grown in sand culturereceiving all nutrients as a complete nutrient solution containing1.5 mMNH₄NO₃, and subjected to one of three defoliation treatments:undefoliated, defoliated on one occasion, or defoliated weekly.¹⁵Nlabelling was used to determine the rate of N uptake, allowingthe amount of N remobilized from storage for the growth of thetwo youngest leaves (subsequently referred to as ‘newleaves’) growing over a 14 d period after defoliationto be calculated. The total plant N uptake by both species wasreduced, compared with undefoliated plants, by both a singleand repeated defoliation, although neither caused complete inhibitionof uptake. Regularly defoliatedL. perennehad a greater reductionin root mass, concomitant with a greater increase in N uptakeper g root than did regularly defoliatedF. rubra. In both species,the amount of N derived from uptake recovered in the new leaveswas unaffected by the frequency of defoliation. BothL. perenneandF.rubramobilized nitrogen to the new leaves after a single defoliation,mobilization being sufficient to supply 50 and 41%, respectively,of the total nitrogen requirement. In regularly defoliated plants,no significant nitrogen was mobilized to the new leaves inL.perenne, and only a small amount was mobilized inF. rubra. Plantsachieved greater leaf regrowth when only defoliated once. Weconclude that increasing the frequency of defoliation of bothL.perenneandF. rubrahad little effect on the uptake of nitrogenby roots which was subsequently supplied to new leaves, butdepleted their capacity for nitrogen remobilization, resultingin a reduction in the rate of growth of new leaves. Lolium perenne; Festuca rubra; defoliation frequency; mobilization; root uptake; nitrogen     

Changes in Free Amino Compounds during the Development of Two Wheat Cultivars Subjected to Different Degrees of Water Stress

The changes of total nitrogen and free amino compounds, exceptproline, in leaves, stems, roots and ears of the wheat cultivarsYecora and Generoso were followed during the development inthe field with and without irrigation. The time course of thesum of the amino compounds was similar in stems and roots, butwas different in the leaves and ears. The levels of particularcompounds for a given cultivar varied with plant development,organ and water stress. Glutamate, alanine, aspartate, and serinewere found at high concentrations in all organs, although theirproportion was substantially lower in the stems and roots whereglutamine and asparagine were also detected in considerableamounts. Large amounts of glutamine were also found in the ears.The concentrations of some compounds increased in the periodsof more severe water stress, although the concentration of totalnitrogen did not show a similar response. From correlationsbetween      

Seasonal Changes of the Metabolites in the Leaves, Bark and Xylem Tissues of Olive Tree (Olea europaea. L) I. Nitrogenous Compounds

The changes of total nitrogen and free amino compounds wereexamined in leaves, bark and xylem tissues of olive tree duringa complete annual cycle. In leaves total nitrogen decreasedin spring/early-summer but reached the highest level in autumn-winterperiods. In bark tissues total nitrogen fluctuations were mainlyrelated to the periodical cambial activities. In xylem tissuestotal nitrogen was detected in low levels with no fluctuationsover the year. The free amino compounds in leaves showed seasonalpeaks during the spring and autumn stem elongations, while aconsiderable restriction of their levels was detected in summer.Glutamate, aspartate, proline, alanine, serine and -aminobutyrateare the predominant amino acids detected in leaves. Glutamateand aspartate increased considerably in spring and autumn periodsbut disappeared in summer. Proline remained almost unchangedover the year. Alanine, in addition to the spring and autumnfluctuations, also fluctuated in summer, -aminobutyrate wasdetected at highest levels in winter. In bark tissues, the patternof fluctuations and the composition of the free amino compoundswere similar to those of leaves. In xylem tissues free aminocompounds were detected in high levels over the year exceptfor the drops in spring and summer cambial activity periods.Xylem was the most important reservoir for the readily transportablesoluble nitrogen being accumulated during the maturing of theproduced new xylem. Arginine, glutamine, alanine, glutamate,aspartate, serine, -aminobutyrate and proline are the most prevalentamino compounds in xylem. Arginine and glutamine showed extensiveinterchanges. Arginine increased in autumn while glutamine andalanine showed low levels in the same period. Olive tree (Olea europaea L), amino acids     

Effects of Edaphic Factors on the Soluble Carbohydrate Content of Roots of Lolium perenneL. and Trifolium repens L.

In a 3³x2² factorial experiment with Lolium perenne L. and Trifoliumrepens L. grown at three soil moisture tensions, three levelsof nitrogen and three levels of phosphorus, there was a widerange of percentage soluble carbohydrate (TSC) in the roots.The significant positive correlations between percentage ofTSC and root weight suggest that root growth-rate was proportionalto the concentration of TSC in the roots. In other experiments with Lolium perenne grown at five constantsoil temperatures and factorial combinations with three soilmoisture treatments with and without manure, there was declinein the percentage of TSC with increasing temperature. This impliedthat the respiration sink strength in the roots did not controlthe partitioning of photosynthate to the roots. No firm conclusions can be drawn from this work, but it is inferredthat there is a dynamic balancing mechanism in the foliage whichcontrols the partitioning of photosynthate between foliage androot growth. This mechanism appears to divert carbohydrate toroots in inverse proportion to root activity.     

Changes in Total Nitrogen and Free Amino Acids in Stem Cuttings of Mulberry (Morus alba L.)

Changes in total nitrogen and free amino acid contents in stemcuttings of Morus alba have been studied. The fresh and dryweights and total nitrogen amounts of the parent stems of cuttingsdecreased initially after cutting. Their increase follows theformation of main roots in cuttings, suggesting that, like carbohydrates,sugars and starch, stored nitrogenous substances are used forsprouting and rooting of cuttings. Amino acids found in stems,roots and shoots are those common in other higher plants withthe exception of pipecolic acid and 5-hydroxypipecolic acid.Significant changes in the levels of asparagine, proline, arginine,-aminobutyric acid and alanine in roots, bark and wood of parentstems were observed during cutting growth, whereas those ofother amino acids remained comparatively constant; the mostpredominant amino acid in the starting materials was proline.while that in the cuttings during growth was asparagine. Theresults suggest that, among free amino acids, asparagine, prolineand arginine play the major part in storage of nitrogen in mulberry.The importance of glut-amine and asparagine in nitrogen metabolismin mulberry has been discussed.     

Characterization of nitrogen relationships between Sorghum bicolor and the root-hemiparasitic angiosperm Striga hermonthica (Del.) Benth. using K15 NO3 as isotopic tracer

The role of the host in the nitrogen nutrition of Striga hermonthica (Del.) Benth. (Scrophulariaceae) parasitic on Sorghum bicolor cv. SH4 Arval has been investigated using (15)N-nitrate as the tracer. It is shown that, when nitrate is absorbed only by the roots of the host plant, a rapid transfer of nitrogen to the parasite can be detected. The xylem sap of S. hermonthica contained approximately equal amounts of nitrate and amino acids, mostly glutamine and asparagine. Infection altered the free amino acid profile of the host tissues, leading notably to a large increase in asparagine and a decrease in glutamine. The haustoria of S. hermonthica, although rich in nitrate, showed a low concentration of free amino acids, particularly lacking in asparagine and glutamine. The roots of S. hermonthica, in contrast, were rich in both asparagine and glutamine while, in the shoots, asparagine constituted 80% of the total FAA pool. Asparagine was also found to be the primary (15)N-enriched amino acid in the shoots of S. hermonthica while, interestingly, it was glutamate that was most strongly enriched in the roots. It is concluded that nitrogen nutrition in S. hermonthica is based on a supply of both nitrate and amino acids from the host. This implies a non-specific transfer in the transpiration stream. Nitrate reduction probably occurs mainly in the leaves of the parasite. Assimilation also occurs in S. hermonthica and excess nitrogen is stored as the non-toxic nitrogen-rich compound, asparagine. This specific trait of nitrogen metabolism of the parasite is discussed in relation to the effect of nitrogen fertilization on reducing infestation.     

Assimilation of Nitrogen in Mutants Lacking Enzymes of the Glutamate Synthase Cycle

¹⁵N labelling was used to investigate the pathway of nitrogenassimilation in photorespiratory mutants of barley (Hordeumvulgare cv. Maris Mink), in which the leaves have low levelsof glutamine synthetase (GS) or glutamate synthase, key enzymesof ammonia assimilation. These plants grew normally when maintainedin high CO₂, but the deletions were lethal when photorespirationwas initiated by transfer to air. Enzyme levels in roots weremuch less affected, compared to leaves, and assimilation oflabelled nitrate into amino acids of the root showed very littledifference between wild type and mutants. Organic nitrogen wasexported from roots in the xylem sap mainly as glutamine, levelsof which were somewhat reduced in the GS-deficient mutant andenhanced in the glutamate synthase deficient mutant. In theleaf, the major effect was seen in the glutamatesynthase mutant,which had an extremely limited capacity to utilize the importedglutamine and amino acid synthesis was greatlyrestricted. Thiswas confirmed by the supply of [¹⁵N]-glutamine directly to leaves.Leaves of the GS-deficient mutant assimilatedammonia at about75% the rate found for the wild type, and this was almost completelyeliminated by addition of the inhibitormethionine sulphoximine.Root enzymes, together with residual levels of the deleted enzymesin the leaves, have sufficient capacityfor ammonia assimilation,through the glutamate synthase cycle, to provide adequate inputof nitrogen for normal growth of themutants, if photorespiratoryammonia production is suppressed. Key words: Hordeum vulgare, ¹⁵N, glutamine synthetase, glutamate synthase, ammonia assimilation     

Root exudates: a pathway for short-term N transfer from clover and ryegrass

The short-term transfer of nitrogen (N) from legumes to grasses was investigated in two laboratory studies. One study was done in pots where the roots of white clover (Trifolium repens L.) and perennial ryegrass (Lolium perenne L.) were allowed to co-exist, and a second study was performed using a micro-lysimeter system designed to maintain nutrient flow from the clover to the grass, whilst removing direct contact between the root systems. The ¹⁵N-dilution technique was used to quantify the transfer of N between species. Levels of ammonia and amino acids were measured in root exudates. The amounts of N transferred were in the same order of magnitude in both the pot and micro-lysimeter experiments. In the micro-lysimeter experiment, 0.076 mg of N were transferred per plant from clover to ryegrass during the course of the experiment. Ammonium exudation was much higher than amino acid exudation. The most abundant amino acids in both clover and ryegrass root exudates were serine and glycine. However, there was no correlation between the free amino acid profile of root extracts and exudates for both plant species: Asparagine was the major amino acid in clover roots, while glutamine, glutamate and aspartate were the major amino acids in ryegrass roots. Comparison of exudates obtained from plants grown in non-sterile or axenic conditions provides evidence of plant origin of ammonium, serine and glycine.     

The Influence of Nitrogen Nutrition on the Accumulation of Free Amino Acids in Root Tissue of Urtica dioica and their Apical Transport in Xylem Sap

Urtica dioica plants were grown on a nitrogen supply of 3, 15and 22 mM with nitrate and ammonium as nitrogen source. In contrastto nitrate reductions amino acid synthesis occurred in roottissue. At 3 mM ammonium obviously the amino acids were rathertransported via xylem upwards to the shoots than stored in theroots. Particularly increased ammonium supply led to stimulatedstorage of free amino acids in the roots, mainly as asparagineand arginine. In xylem asparagine was the dominant nitrogentransporting compound, while arginine was hardly translocated.With the enhancement of nitrogen supply, the second amide, glutamine,became more and more important with respect to the transportof nitrogen. (Received September 3, 1984; Accepted November 2, 1984)     

Free amino acids and total nitrogen during shoot development in Scots pine seedlings

The concentration of free amino acids and total nitrogen was studied in needles, stems and roots of seedlings of Pinus sylvestris L. for five weeks during the second growth period ("summer"). In one group of seedlings the source/sink relation was disturbed through removal of the terminal buds. The seedlings were cultivated in artificial year-cycles in a climate chamber.
Total nitrogen increased in needles and sterns of intact seedlings in the beginning of the "summer" and decreased during shoot growth. In seedlings, from which the buds had been removed, nitrogen remained at high levels in the primary needles and accumulated in steins and roots. The results are consistent with utilization of nitrogen in older needles and in the stem during shoot elongation.
The pool of free amino acids increased in the beginning of the "summer" and decreased after bud break in primary needles, stems and roots. Arginine and glutamine, in the roots also asparagine, were the dominating amino acids (amides included). Together, these compounds (plus glutamate and aspartate) contributed about 90% of the nitrogen in the amino acid pool in all organs. In primary needles and in the stem, arginine predominated at the end of hardening (75–85% of the amino acid nitrogen). Free amino acids contributed at most ca 10% of the total nitrogen in primary needles, where the ratio of free amino acid nitrogen: total nitrogen was highest at the end of dormancy and in the early "summer". Free amino acids accumulated after bud removal in primary needles and especially in stems and roots. Glutamine became relatively more dominant than arginine in the different organs.
The observations are consistent with the role of arginine and glutamine for storage and transport of nitrogen in conifers. Because of the low concentrations of amino acid nitrogen in the primary needles, arginine is not considered a major nitrogen reserve in needles of Scots pine seedlings.     

The Route of Nitrate-Nitrogen Assimilation in the Root of Datura stramonium L.

Datura roots were pressure-infiltrated with 400 µg ml^–115N-nitrate feeding solutions with and without the additionof 7 mM L-methionine-DL-sulphoximine (MSO), a glutamine synthetaseinhibitor. Over a 30 min time course the main diversion of newlyreduced ¹⁵N in MSO untreated roots was to glutamine. In MSO-treatedroots ammonia assimilation into amino compounds was completelysuppressed, with resultant accumulation of a large ¹⁵N ammoniapool. This treatment also caused marked concentrational changesin the free amino compound pools, suggesting that conditionsof nitrogen stress had been induced. Glutamate dehydrogenaseactivity was unaffected by the MSO treatment. The results are consistent with the concept that the glutaminesynthetase/glutamate synthase pathway is the major route ofnewly reduced nitrogen assimilation in Datura roots.