Effect of ammonium nutrition on the activity of nitrate reductase in the roots of apple seedlings

Active extracts of nitrate reductase were prepared from theroots of apple seedlings c.v. Granny Smith which were grownin nutrient solution under controlled enviromental conditions.The nutrient solutions contained various ratios of nitrate andammonium ions but all the treatments contained a total of 112ppm nitrogen. Maximum nitrate reductase activity in the roots was obtainedwhen plants were supplied with nitrate as the sole source ofnitrogen. Roots grown in solution containing only ammonium nitrogenhad little or no activity. When plants were supplied with bothforms of nitrogen in the nutrient solution, the presence ofammonium ions markedly lowered the activity of nitrate reductasein the roots. Plants supplied with 98 ppm nitrate nitrogen plus14 ppm ammonium nitrogen had activities only half those of plantsgrown in nitrate alone. Plants supplied with equal amounts ofammonium and nitrate nitrogen had activities less than one sixththose of plants grown in nitrate alone. (Received June 3, 1972; )     

Preferential Assimilation of Ammonium Ions from Ammonium Nitrate Solutions by Apple Seedlings

Apple seedlings, Pyrus malus L., were grown in complete nutrient solutions containing nitrate, ammonium, or ammonium plus nitrate as the nitrogen source. Uptake of nitrogen was calculated from depletion measurements of the nutrient solutions and by using ¹⁵N labelled nitrate and ammonium salts. If the plants received nitrogen as ammonium only or as nitrate only, the amounts of nitrogen taken up were similar. However, if the seedlings were supplied with ammonium nitrate, the amount of nitrate-nitrogen assimilated was only half that of ammonium. Nevertheless, if ammonium and nitrate were supplied to a plant with a split-root system, with each root half receiving a different ion, the uptakes were similar. The possibility of independent inhibition by ammonium of both nitrate uptake and reduction in the roots is discussed.     

Inhibition of ammonium assimilation restores elongation of seminal rice roots repressed by high levels of exogenous ammonium

Elongation of seminal and lateral roots of rice seedlings was markedly inhibited by high ammonium levels in growth medium. However, high exogenous nitrate concentrations had little inhibitory effect on root growth. The objective of this study was to elucidate the relationship between inhibition of rice root growth induced by high ammonium conditions and ammonium assimilation in the seedlings. Activity of glutamine synthetase (GS) was kept at a low level in the seminal roots of the seedlings grown under high nitrate levels. In contrast, high ammonium levels significantly enhanced the GS activity in the roots, so that Gln abundantly accumulated in the shoots. These results indicate that ammonium assimilation may be activated in the seminal roots under high ammonium conditions. Application of methionine sulfoximine (MSO), an inhibitor of GS, relieved the repression of the seminal root elongation induced by high ammonium concentrations. However, the elongation of lateral roots remained inhibited even under the same condition. Furthermore, MSO drastically increased ammonium level and remarkably decreased Gln level in the shoots grown under high ammonium conditions. These results show that, for rice seedlings, an assimilatory product of ammonium, and not ammonium itself, may serve as an endogenous indicator of the nitrogen status involved in the inhibition of seminal root elongation induced by high levels of exogenous ammonium.     

Cycling of amino-nitrogen between shoots and roots in wheat seedlings

Summary One part of a split root system of wheat seedlings received full nutrient solution with¹⁵N-nitrate, the other received an identical solution with unlabelled nitrate. Appearance of labelled amino compounds was measured in the xylem sap exuding from roots not supplied directly with¹⁵N-nitrate after removal of the¹⁵N-nitrate-fed roots. This material indicates cycling of nitrogen from the shoots and through the roots. About 60 per cent of the nitrogen in the xylem appears to be cycling in this way.     

Contamination of Ammonium-Based Nutrient Solutions by Nitrifying Organisms and the Conversion of Ammonium to Nitrate

Conversion of ammonium to nitrate and contamination by nitrifying organisms are often assumed not to be significant in ammonium-based nutrient solutions. To assess this assumption, maize (Zea mays) and pea (Pisum sativum) were grown under greenhouse conditions in aeroponic, hydroponic, and sand-culture systems containing 2 mM ammonium chloride as the sole nitrogen source and evaluated for the activity of contaminating nitrifying organisms. In all three culture systems, root colonization by nitrifying organisms was detected within 5 d, and nitrate was detected in the nutrient solution within 10 d after seedling transfer. In sand culture, solution nitrate concentration reached 0.35 mM by the end of the 17-d experiment. Consistent with the microbial ammonium oxidation sequence, nitrite was detected earlier than nitrate and remained at lower levels throughout the experiment. Nitrate was found in significant quantities in root and shoot tissues from seedlings grown in ammonium-based nutrient solutions in all of the solution culture systems. Maize seedlings grown in an ammonium-based hydroponic system contained nitrate concentrations at 40% of that found in plants grown in nitrate-based solution. Determination of nitrate (or nitrite) levels in the nutrient solution was the weakest indicator of the activity of nitrifying organisms. A bioassay for the presence of nitrifying organisms in combination with tissue analysis for nitrate was a better indicator of microbial conversion of ammonium to nitrate in nutrient solution culture. The results have implications for the use of ammonium-based nutrient solutions to obtain plants suitable for research on induction of nitrate uptake and reduction or for research using solution culture to compare ammonium versus nitrate fertilization.     

The influence of ammonia in nutrient solution on growth and metabolism of cucumber plants

Summary Shoot yield of cucumber plants grown 18 days in nutrient solution with 0.06 mM NH₃ was decreased. Root yield was diminished at 0.09 mM NH₃ The ammonia treatment caused heavy chlorosis increasing with age of leaves. This chlorosis was not due to any nutrient deficiency. Ammonia also influenced the morphology of roots. They were clearly shorter caused by a much smaller size of root cells.The decrease of yield was linked to a reduction of assimilation occurring not only after a long influence of ammonia lasting 14 days, but also within one hour after starting the NH₃ treatment. The decline of assimilation was probably caused by a higher resistance of stomata against CO₂ influx in leaf tissue as can be concluded from the observation that transpiration was decreased in the same way as assimilation.The effect of ammonia in nutrient solution could also be due to the occurrence of higher NH₃ concentrations in leaf tissue, because both, pH of plant press sap as well as NH₄ concentration of plant tissue, were increased.Furthermore, it is shown that the nitrate content of plant tissue was diminished by ammonia whereas ammonium and amide content were raised. Regulation of nitrate uptake of plants by means of ammonium and amide content of tissue is discussed.     

Distribution of Nitrogen during Growth of Sunflower (Helianthus annuus L.)

The accumulation, distribution and redistribution of dry matterand nitrogen is described for Helianthus annuus L. cv. Hysun21 grown on 6 mM urea in glasshouse culture. Seed dry matterand nitrogen were transferred to seedlings with net efficienciesof 40 and 86 per cent respectively. At flowering, the stem hadmost of the plant's dry matter and the leaves most of its nitrogen.About 35 per cent of the plant's nitrogen accumulated afterthree-row anthesis. The amount of protein in vegetative parts,especially leaves, declined after flowering. Concentrationsof free amino compounds also decreased during growth. Matureseeds had 38 per cent of the total plant dry weight and 68 percent of the total nitrogen. Seeds acquired 33 per cent of theirdry matter and nitrogen from redistribution from above-groundplant parts. The stem was most important for storage of carbohydrate,leaves the most important for nitrogen. Over 50 per cent ofthe nitrogen in the stem and leaves was redistributed. Plantsthat received 6 mM nitrate accumulated more dry matter thanurea-grown plants. Seeds from nitrate-grown plants were heavier(58 mg) than those of urea-grown plants (46 mg), and their percentageoil was greater (50 and 41 respectively). The amount of nitrogenper seed was the same. Little or no urea was detected in xylem sap of plants suppliedwith 5 mM urea, but it was detected in sap of plants which received25 mM. Concentrations of urea and amino compounds in the sapdecreased up the stem. Plants supplied with nitrate had mostof the nitrogen in xylem sap as NO₂^–, suggesting littlenitrate reduction in roots. Plants grown on 6 mM nitrate andchanged to high levels of urea-nitrogen for 14 days still hadhigh levels of nitrate; little nitrate remained in plants receivinglow levels of urea. When urea is applied in irrigation waterto field-grown sunflower, the nitrogen is subsequently takenup as nitrate due to rapid nitrogen transformations in the soil. Helianthus annuus L., sunflower, urea, nitrate, nitrogen transport, xylem sap, nitrogen accumulation nitrogen distribution     

Local NO3- or NH4+ supply modifies the root system architecture of Cedrus atlantica seedlings grown in a split-root device

To study the effects of local nitrate or ammonium supply on the architecture of the Cedrus atlantica root system, cedar seedlings were grown in split-root boxes in a growth chamber. In each box-compartment, roots were fertilized with a solution containing nitrogen, either as nitrate [Ca(NO₃)₂] or ammonium (NH₄Cl), supplied at 0.1 or 5.0 mM. For each seedling, the shoot growth was measured twice a week for 3 months. The root system architecture was also recorded twice a week by tracing the root elongation through the transparent face of the root observation boxes. The apical diameter of the tap-root relay and that of a representative sample of lateral roots were recorded once a month using a monocular magnifier.

The increase of ammonium or nitrate concentration in the nutrient solution has significantly enhanced the production of lateral roots on the tap-root relay. After 90 days of culture, percentages of short lateral roots obtained with nitrate were higher than those obtained using ammonium. A preferential carbon allocation to the shoots was also obtained with an increasing nitrogen supply. Until the 40th day of culture, the elongation of lateral roots was similar for all treatments and ranged from 0.25 to 0.5 cm day⁻¹. From the 40th day to the 95th day, significant differences were observed between the compared modes and maximum elongation rates were obtained with 5 mM NH₄⁺ (2.18 cm day⁻¹) and 5 mM NO₃⁻ (1.18 cm day⁻¹). Local applications of nitrate and ammonium at a low or a high concentration had local effects on elongation and branching of the root system in the fertilized compartment. Contrasting effects of ammonium and nitrate were observed on the apical diameter of tap-roots and lateral roots. The root-split culture device confirmed that nitrate had local effects on the architecture of the C. atlantica root system.     

Importance of seed Zn content for wheat growth on Zn-deficient soil

Eggplants (Solanum melongena L. cv. Bonica) were grown in a glasshouse during summer under natural light with one unbranched shoot or one shoot with 3 to 4 branches and with or without fruit in quartz sand buffered and not buffered with 0.5% CaCO₃ (w : v), respectively. Nutrient solutions supplied contained nitrate or ammonium as the sole nitrogen source. Compared with nutrient solutions containing nitrate (10 mM), solutions containing ammonium (10 mM) caused a decrease in net photosynthesis of eggplants during early stages of vegetative growth when grown in quartz sand not buffered with CaCO₃. The decrease was not observed before leaves showed interveinal chlorosis. In contrast, net photosynthesis after bloom at first increased more rapidly in eggplants supplied with ammonium than with nitrate nitrogen. However, even in this case, net photosynthesis decreased four weeks later when ammonium nutrition was continued. The decrease was accompanied by epinasty and interveinal chlorosis on the lower leaves and later by severe wilting, leaf drop, stem lesions, and hampered growth of stems, roots, and fruits. These symptoms appeared later on plants not bearing fruits than on plants bearing fruits. If nutrient solutions containing increasing concentrations of ammonium (0.5–30 mM) were supplied after the time of first fruit ripening, shoot growth and set of later flowers and fruits were promoted. In contrast, vegetative growth and reproduction was only slightly affected by increasing the concentration of nitrate in the nutrient solutions. In quartz sand buffered with CaCO₃ ammonium nutrition caused deleterious effects only under low light conditions (shade) and on young plants during rapid fruit growth. If eggplants were supplied with ammonium nitrogen before bloom, vegetative growth was promoted, and set of flowers and fruit occurred earlier than on plants supplied with nitrate. Furthermore, the number of flowers and fruit yield increased. These effects of ammonium nutrition were more pronounced when plants were grown with branched shoots than with unbranched shoots. The results indicate that vegetative and reproductive growth of eggplants may be manipulated without causing injury to the plants by supplying ammonium nitrogen as long as the age of the plants, carbohydrate reserves of the roots, quantity of ammonium nitrogen supplied, and pH of the growth medium are favourable. T W Rufty Section editor     

The effect of NO3- and NH4 + ions on enzymes involved in nitrogen assimilation inPisum sativum L

Nitrate reductase level in leaves of pea plants is higher than in roots despite of the lower content of endogenous nitrate. Addition of ammonium ions to nutrient solution containing nitrate decreases nitrate reductase level in leaves estimatedin vivo while its level estimatedin vitro is increased. Glutamine synthetase (GS) level in roots decreases during short (24 and 48 h) and long (14 d) term cultivation of seedlings in solutions containing ammonium ions. This decrease occurs in leaves only after the long term influence of ammonium ions. Level of this enzyme is higher in plants grown in the presence of nitrogen (ammonium and nitrate) as compared to those grown without the nitrogen. Level of glutamate dehydrogenase in roots is increased after both short and long term cultivation of plants in the presence of ammonium ions.     

Increase in glutamate synthase (NADH) activity in maize seedlings in response to nitrate and ammonium nitrogen

Roots and leaves of Zea mays L. cv. Ganga Safed-2 seedlings grown with nutrient solution containing either 10 m M KNO₃ or NH₄Cl or 5 m M NH₄NO₃ had considerably higher glutamate synthase (NADH, EC 1.4.1.14) activity than the corresponding organs from seedlings grown without any nitrogen. The supply of inorganic nitrogen for a short time, i.e. 3 h, to roots and leaves excised from seedlings grown without nitrogen also increased the enzyme activity in these organs. This increase was more pronounced with nitrate than with ammonium nitrogen. When excised roots and leaves from NH₄NO₃-grown seedlings were incubated in a minus nitrogen medium for 24 h, the enzyme activity declined considerably. This decline was inhibited to some extent by nitrogen, especially by nitrate. Inorganic nitrogen prevented similarly the decline in in vitro enzyme activity during 24 h storage at 25°C, more regularly for the root than for the leaf enzyme. The experiments demonstrate the role of inorganic nitrogen in the regulation of glutamate synthase activity.     

Influence of ammonium on fine root development and rhizosphere pH of Douglas-fir seedlings in sand

Ammonium sulphate is a major component of the air pollutants deposited on forests in the Netherlands. Different amounts of NH₄ ⁺ were added to Douglas-fir seedlings grown in tall containers of sand, to study the influence of high concentrations of NH₄ ⁺ in the soil on the development of fine roots and the effects of nitrogen uptake on rhizosphere pH. At the end of this eight-month experiment part of the ammonium appeared to have nitrified into nitrate. High doses of ammonium negatively affected root length and root length per unit of dry matter (specific root length). Although Douglas fir shows a preferential ammonium uptake in nutrient solutions the increases in the pH of the rhizosphere in this experiment indicate that nitrogen was mostly taken up as nitrate. When the ammonium concentration in the soil is low, it cannot be taken up readily because of its low mobility in soil. Shoot growth was stimulated by high availability of nitrogen. The possible effects of high doses of ammonium on long-term forest vitality are discussed.     

Effects of seed nickel reserves or externally supplied nickel on the growth,nitrogen metabolites and nitrogen use efficiency of urea- or nitrate-fed soybean

Background and aims

Nickel (Ni) has a critical role in the urea metabolism of plants. This study investigated the impact of seed Ni content along with external Ni supply on the growth, various nitrogen (N) metabolites and N use efficiency (NUE) of soybean plants.

Methods

Soybean plants raised from Ni-poor or Ni-rich seeds were grown in nutrient solution with or without external Ni supply and fed with either urea or nitrate as the sole N source. The changes in growth, leaf chlorophyll levels, Ni and N concentrations of different plant parts, tissue accumulation of various N metabolites and N uptake of soybean as well as NUE and its components were examined.

Results

Nickel starvation reduced the shoot biomass of urea-fed plants by 25 % and the leaf chlorophyll levels by up to 35 %, but nitrate-fed plants were unaffected. Visual toxicity symptoms were not observed in urea-fed plants. Under urea supply, Ni-deficient plants had lower levels of total N, protein and free amino acids in various organs. Root uptake of urea was severely depressed in Ni-deprived plants. Availability of Ni did not have any effect on the NUE of nitrate-fed plants, whereas its deficiency reduced the NUE of urea-fed plants by 30 %. The growth and N nutritional status of urea-fed soybean were significantly improved by high seed Ni reserves as well as external Ni supply.

Conclusion

Adequate Ni supply is required for maximizing the growth, root uptake of urea and NUE of urea-fed plants. Seed Ni reserves contribute significantly to the Ni and thus N nutritional status of soybean.     

Effect of ammonium salt on enzymes of ammonium assimilation in maize seedlings

Glutamate dehydrogenase (GDH E.C. 1.4.1.2.4), glutamine synthetase (GS E.C. 6.3.1.2) and glutamate synthase (glutamine oxoglutarate amino transferase, GOGAT E.C. 2.6.1.53) activities, protein and organic nitrogen contents and growth of roots and shoots of maize seedlings raised in dark at 25±2°C in half strength Hoagland’s solution containing different ammonium salts as source of nitrogen, were determined to assess the contribution of alternate pathways in ammonium assimilation. Ammonium nitrate or in some cases ammonium chloride appeared to be the best source for both root and shoot growth and for increase in protein, total nitrogen and the enzymes of ammonium assimilation. In roots, NH₄-nitrogen appeared to be assimilated by both GDH as well as GS-GOGAT pathways specially in the dark grown seedlings, while in shoots it was primarily by GS-GOGAT pathway.     

A novel morphological response of maize (Zea mays) adult roots to heterogeneous nitrate supply revealed by a split‐root experiment

Approximately 35–55% of total nitrogen (N) in maize plants is taken up by the root at the reproductive stage. Little is known about how the root of an adult plant responds to heterogeneous nutrient supply. In this study, root morphological and physiological adaptations to nitrate‐rich and nitrate‐poor patches and corresponding gene expression of ZmNrt2.1 and ZmNrt2.2 of maize seedlings and adult plants were characterized. Local high nitrate (LoHN) supply increased both lateral root length (LRL) and density of the treated nodal roots of adult maize plants, but only increased LRL of the treated primary roots of seedlings. LoHN also increased plant total N acquisition but not N influx rate of the treated roots, when expressed as per unit of root length. Furthermore, LoHN markedly increased specific root length (m g^?1) of the treated roots but significantly inhibited the growth of the lateral roots outside of the nitrate‐rich patches, suggesting a systemic carbon saving strategy within a whole root system. Surprisingly, local low nitrate (LoLN) supply stimulated nodal root growth of adult plants although LoLN inhibited growth of primary roots of seedlings. LoLN inhibited the N influx rate of the treated roots and did not change plant total N content. The gene expression of ZmNrt2.1 and ZmNrt2.2 of the treated roots of seedlings and adult plants was inhibited by LoHN but enhanced by LoLN. In conclusion, maize adult roots responded to nitrate‐rich and nitrate‐poor patches by adaptive morphological alterations and displayed carbon saving strategies in response to heterogeneous nitrate supply.     

Root exudation and rhizoplane bacterial abundance of barley (Hordeum vulgare L.) in relation to nitrogen fertilization and root growth

The abundance of bacteria in the rhizoplane of barley varieties was investigated at different soil nitrogen levels. Increased amendments of nitrogen resulted in higher bacterial numbers in the rhizoplane of barley seedlings of different varieties. A negative correlation was found between nitrogen level in the soil and the growth rate of the seedling roots. The effect of nitrogen on the bacterial abundances could be indirect through changed root growth and thereby changed exudation. The exudation of soluble organic carbon componds from barley seedling roots were measured in hydroponic culture. The effect of natural variation in root growth rate and of different concentrations of nitrogen in the nutrient solution was investigated. The amount of exudates consituted 2–66% of the dry weight increase in root biomass, depending on the root growth. Slower growing roots released considerably more organic carbon per unit root weight than faster growing roots. The variation in root exudation appeared to be mainly explained by differences in root growth, rather than of the nitrogen concentration in the nutrient solution. A significantly higher exudation rate was found during day time compared to night.     

Accumulation of Oxalate in Tissues of Sugar-beet, and the Effect of Nitrogen Supply

In field-grown sugar-beet concentration of insoluble oxalatewas low in roots and high (about 12 per cent of ethanol insolublematerial) in leaves, and for a particular leaf the concentrationincreased continuously during its life. Of the insoluble oxalate,15–30 per cent was present as the magnesium salt and theremainder as the calcium salt. Oxalate contents of plants grownin culture solutions with nitrate as nitrogen source were similarto those of plants grown in soil, but when nitrogen was suppliedas ammonium sulphate or ammonium nitrate both soluble and insolubleoxalate were low. Plants grown in soil with regular additionsof ammonium sulphate or ammonium nitrate also had very low concentrationsof soluble oxalate although insoluble oxalate was only slightlylower than with nitrate nitrogen. Disks of root or leaf tissuewashed for several days in distilled water lost insoluble oxalatebut when washed in tap water insoluble oxalate increased morethan twofold. Addition of calcium and nitrate to the distilledwater caused an increase of insoluble oxalate, while additionof potassium caused a decrease. Use of ¹⁴C labelled oxalateand washing experiments showed that oxalate can be metabolizedby tissue disks and so is not necessarily a final product ofmetabolism. The accumulation of oxalate appears to be connectedwith the assimilation of nitrate and the preservation of thecation-anion balance of the plant.     

Nitrogen Starvation and the Regulation of Glutamine Synthetase in Agmenellum quadruplicatum

The level of glutamine synthetase activity in Agmenellum quadruplicatum strain PR-6 was dependent on the nitrogen source used for growth and on the nutritional status of the cells. During exponential growth, glutamine synthetase activity was low in cells grown on ammonia, urea, or nitrate. During the transition from nitrogen replete to nitrogen starved growth, glutamine synthetase activity began to rise. With ammonia as a nitrogen source, glutamine synthetase activity as determined in whole cells increased from 1 nanomole per minute per milliliter during exponential growth to 22 nanomoles per minute per milliliter during severe nitrogen starvation. In cells grown on nitrate the increase was from 5 to 39 nanomoles per minute per milliliter, and in cells grown on urea the increase was from 4 to 31 nanomoles per minute per milliliter.     

Functional aspects of root architecture and mycorrhizal inoculation with respect to nutrient uptake capacity

The aim of this research was to investigate the effect of arbuscular mycorrhizal (AM) colonisation on root morphology and nitrogen uptake capacity of carob ( Ceratonia siliqua L.) under high and low nutrient conditions. The experimental design was a factorial arrangement of presence/absence of mycorrhizal fungus inoculation ( Glomus intraradices) and high/low nutrient status. Percent AM colonisation, nitrate and ammonium uptake capacity, and nitrogen and phosphorus contents were determined in 3-month-old seedlings. Grayscale and colour images were used to study root morphology and topology, and to assess the relation between root pigmentation and physiological activities. AM colonisation lead to a higher allocation of biomass to white and yellow parts of the root. Inorganic nitrogen uptake capacity per unit root length and nitrogen content were greatest in AM colonised plants grown under low nutrient conditions. A better match was found between plant nitrogen content and biomass accumulation, than between plant phosphorus content and biomass accumulation. It is suggested that the increase in nutrient uptake capacity of AM colonised roots is dependent both on changes in root morphology and physiological uptake potential. This study contributes to an understanding of the role of AM fungi and root morphology in plant nutrient uptake and shows that AM colonisation improves the nitrogen nutrition of plants, mainly when growing at low levels of nutrients.     

Ammonium and nitrate nutrition inPlantago lanceolata L. andPlantago major L. ssp.major

With the aims (1) to test whether the different natural occurrence of twoPlantago species in grasslands is explained by a different preference of the species for nitrate or ammonium; (2) to test whether the different occurrence is explained by differences in the flexibility of the species towards changes in the nitrogen form; (3) to find suitable parameters as a tool to study ammonium and nitrate utilization of these species at the natural sites in grasslands, plants ofPlantago lanceolata andP. major ssp.major were grown with an abundant supply of nitrate, ammonium or nitrate+ammonium as the nitrogen source (0.5 mM). The combination of ammonium and nitrate gave a slightly higher final plant weight than nitrate or ammonium alone. Ammonium lowered the shoot to root ratio inP. major. Uptake of nitrate per g root was faster than that of ammonium, but from the mixed source ammonium and nitrate were taken up at the same rate. In vivo nitrate reductase activity (NRA) was present in both shoot and roots of plants receiving nitrate. When ammonium was applied in addition to nitrate, NRA of the shoot was not affected, but in the root the activity decreased. Thus, a larger proportion of total NRA was present in the shoot than with nitrate alone. In vitro glutamate dehydrogenase activity (GDHA) was enhanced by ammonium, both in the shoot and in the roots.In vitro glutamine synthetase activity (GSA) was highest in roots of plants receiving ammonium. Both GDHA and GSA were higher inP. lanceolata than inP. major. The concentration of ammonium in the roots increased with ammonium, but it did not accumulate in the shoot. The concentration of amino acids in the roots was also enhanced by ammonium. Protein concentration was not affected by the form of nitrogen. Nitrate accumulated in both the shoot and the roots of nitrate grown plants. When nitrate in the solution was replaced by ammonium, the nitrate concentration in the roots decreased rapidly. It also decreased in the shoot, but slowly. It is concluded that the nitrogen metabolism of the twoPlantago species shows a similar response to a change in the form of the nitrogen source, and that differences in natural occurrence of these species are not related to a differential adaptation of nitrogen metabolism towards the nitrogen form. Suitable parameters for establishing the nitrogen source in the field are thein vivo NRA, nitrate concentrations in tissues and xylem exudate, and the fraction of total reduced nitrogen in the roots that is in the soluble form, and to some extent thein vitro GDHA and GSA of the roots. Grassland Species Research Group. Publ. no 118.