Metabolic Pathway of alpha-Ketoglutarate in Citrus Leaves as Affected by Phosphorus Nutrition

The uptake and metabolism of α-[5-¹⁴C]ketoglutarate by phosphorus-deficient and full nutrient (control) lemon (Citrus limon) leaves were studied over various time intervals. After 45 minutes in P-deficient leaves, the bulk of incorporated ¹⁴C appeared in organic acids and much less in amino acids, while in the control leaves, the ¹⁴C contents of organic and amino acids were equal. In P-deficient leaves, after longer incubation times the ¹⁴C content of organic acids and amino acids increased, while that of CO₂ and residue fractions remained low. In full nutrient leaves the ¹⁴C content of amino acids and organic acids decreased after longer incubation time and increased in the insoluble residue and CO₂. In full nutrient leaves the organic and amino acid metabolism were closely related and accompanied by protein synthesis and CO₂ release, while in P-deficient leaves an accelerating accumulation of arginine and citric acid was linked together with inhibition of protein synthesis and CO₂ liberation.     

Trehalose 6–phosphate coordinates organic and amino acid metabolism with carbon availability

Trehalose 6–phosphate (Tre6P) is an essential signal metabolite in plants, linking growth and development to carbon metabolism. The sucrose–Tre6P nexus model postulates that Tre6P acts as both a signal and negative feedback regulator of sucrose levels. To test this model, short‐term metabolic responses to induced increases in Tre6P levels were investigated in Arabidopsis thaliana plants expressing the Escherichia coli Tre6P synthase gene (otsA) under the control of an ethanol‐inducible promoter. Increased Tre6P levels led to a transient decrease in sucrose content, post‐translational activation of nitrate reductase and phosphoenolpyruvate carboxylase, and increased levels of organic and amino acids. Radio‐isotope (¹⁴CO₂) and stable isotope (¹³CO₂) labelling experiments showed no change in the rates of photoassimilate export in plants with elevated Tre6P, but increased labelling of organic acids. We conclude that high Tre6P levels decrease sucrose levels by stimulating nitrate assimilation and anaplerotic synthesis of organic acids, thereby diverting photoassimilates away from sucrose to generate carbon skeletons and fixed nitrogen for amino acid synthesis. These results are consistent with the sucrose–Tre6P nexus model, and implicate Tre6P in coordinating carbon and nitrogen metabolism in plants.     

Plasma free amino acid kinetics in rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) using a bolus injection of <Superscript>15</Superscript>N-labeled amino acids

To gain insight into the metabolic design of the amino acid carrier systems in fish, we injected a bolus of ¹⁵N amino acids into the dorsal aorta in mature rainbow trout (Oncorhynchus mykiss). The plasma kinetic parameters including concentration, pool size, rate of disappearance (R _d), half-life and turnover rate were determined for 15 amino acids. When corrected for metabolic rate, the R _d values obtained for trout for most amino acids were largely comparable to human values, with the exception of glutamine (which was lower) and threonine (which was higher). R _d values ranged from 0.9 μmol 100 g⁻¹ h⁻¹ (lysine) to 22.1 μmol 100 g⁻¹ h⁻¹ (threonine) with most values falling between 2 and 6 μmol 100 g⁻¹ h⁻¹. There was a significant correlation between R _d and the molar proportion of amino acids in rainbow trout whole body protein hydrolysate. Other kinetic parameters did not correlate significantly with whole body amino acid composition. This indicates that an important design feature of the plasma-free amino acids system involves proportional delivery of amino acids to tissues for protein synthesis.     

Partitioning of Photosynthetically Fixed 14CO2 Into Oil and Curcumin Accumulation in Curcuma Longa Grown Under Iron Deficiency

Changes in leaf growth, photosynthetic efficiency, and incorporation pattern of photosynthetically fixed ¹⁴CO₂ in leaves 1 and 2 from plant apex, in roots, and rhizome induced in Curcuma by growing in a solution culture at Fe concentration of 0 and 5.6 g m^–3 were studied. ¹⁴C was incorporated into primary metabolites (sugars, amino acids, and organic acids) and secondary metabolites (essential oil and curcumin). Fe deficiency resulted in a decrease in leaf area, its fresh and dry mass, chlorophyll (Chl) content, and CO₂ exchange rate at all leaf positions. The rate of ¹⁴CO₂ fixation declined with leaf position, maximum being in the youngest leaf. Fe deficiency resulted in higher accumulation of sugars, amino acids, and organic acids in leaves at both positions. This is due to poor translocation of metabolites. Roots and rhizomes of Fe-deficient plants had lower concentrations of total photosynthate, sugars, and amino acids whereas organic acid concentration was higher in rhizomes. ¹⁴CO₂ incorporation in essential oil was lower in the youngest leaf, as well as incorporation in curcumin content in rhizome. Fe deficiency influenced leaf area, its fresh and dry masses, CO₂ exchange rate, and oil and curcumin accumulation by affecting translocation of assimilated photosynthates.     

Export of amino acids to the phloem in relation to N supply in wheat

The effect of different N supply on amino acid export to the phloem was studied in young plants of wheat (Triticum aestivum L. cv. Klein Chamaco), using the exudation in EDTA technique. Plants were grown in a growth cabinet in pots with sand, and supplied with nutrient solutions of different NO₃^? concentrations. When plants were grown for 15 days with nutrient solutions containing 1.0, 3.0, 5.0, 10.0, 15.0 or 20.0 mM KNO₃, the exudation rate of sugars from the phloem was unaffected by N supply, but sugars accumulated in the leaf tissue when the N supply was limiting for growth. On the other hand, the rate of exudation of amino acids was proportional to the NO₃^? concentration in the nutrient solution. When the supply of N to plants grown for 15 days with 15.0 mM NO₃^? was interrupted, the exudation of sugars was again unaffected, but there was a fast decrease in the amount of amino acids exudated, and of the concentration of amino acids and nitrogen in the tissues. Also, when 10-day-old plants grown without N were supplied with 15.0 mM NO₃^?, there was a sharp increase in the exudation of amino acids, without changes in the amount of sugar exudated. The rate of exudation of amino acids to the phloem was independent of the concentration of free amino acids in the leaves in all three types of experiment. Asp was the most abundant amino acid in the leaf tissue, while Glu was the one most abundant in the phloem exudate. Asp and Ala were exported to the phloem at a rate lower than expected from their leaf tissue concentrations, indicating some discrimination. On the contrary, Glu showed a preferential export at low N supply. It is concluded that the rate of amino acid export from the leaf to the phloem is dependent on the N available to the plant. This N is used for synthesis of leaf protein when the supply is low, exported to the phloem when supply is adequate, and accumulated in the storage pool when supply is above plant demand.     

氨基酸添加对亚热带森林红壤氮素转化的影响

为探究氨基酸氮形态对亚热带土壤氮素含量及转化的影响,选择建瓯市万木林保护区的山地红壤为对象,采用室内培养实验法,通过设计60%和90%WHC两种土壤含水量并添加不同性质氨基酸,测定了土壤中铵态氮、硝态氮、可溶性有机氮的含量和氧化亚氮的释放量,分析了可溶性有机碳、土壤p H值的大小变化及其与氮素的相互关系。结果表明:与对照处理相比,氨基酸添加显著增加了土壤NH_4~+-N含量并使土壤p H值升高,且在一定程度上解除了高含水量(90%WHC)对NH_4~+-N产生的抑制,其中甲硫氨基酸的效果最为明显。酸性、碱性、中性氨基酸对土壤NO_3~--N含量和N_2O释放影响不显著,但甲硫氨基酸可显著抑制土壤硝化从而导致NH_4~+-N的积累,并在培养前期抑制土壤N_2O产生而在培养后期促进N_2O释放,总体上促进N_2O释放。60%WHC的氨基酸添加处理较90%WHC条件下降低土壤可溶性有机氮的幅度更大。氨基酸对土壤氮素转化的影响与带电性关系较小,而可能与其分解产物密切相关。可见,不同性质氨基酸处理对森林土壤氮素含量及转化存在不同程度的影响,且甲硫氨基酸对土壤氮素转化的影响机理值得深入研究。     

C14 distribution in photosynthate of tomato as influenced by substrate copper and molybdenum level and nitrogen source

Summary The influence of nitrogen source and micronutrient treatment on the apportionment of C¹⁴ in alcohol extracts was studied in tomato. Plants were exposed at several stages of maturity to C¹⁴O₂ and the ethanol-soluble photosynthate fractionated into sugars, organic acids, and amino acids.Ammonium-treated plants had low levels of organic acids at all stages of maturity, suggesting a rapid conversion to amino acid residues. However, the addition of copper in the absence of molybdenum resulted in increased labelling of the organic acid fraction under nitrate nutrition. The amino acid fraction was reduced at some stages of growth under these conditions. This may indicate that copper interferes with the role of molybdenum in the enzymatic reduction of nitrate.Measurement of the total quantity of some individual free amino acids indicated more efficient utilization of these for protein synthesis in the presence of copper. On the other hand, assay of the C¹⁴-activity of individual free amino acids showed that degree of label bore little relation to micro-nutrient treatment.Plant Science Department and Botany Department, University of Connecticut. Scientific Contribution No. 116, Agricultural Experiment Station, University of Connecticut, Storrs. This paper is based on a portion of the senior author's Ph. D. thesis (University of Connecticut).     

Analysis of energetic amino acid metabolism in Acyrthosiphon pisum: A multidimensional approach to amino acid metabolism in aphids

Aphids are highly specialized insects that feed on the phloem-sap of plants, the amino acid composition of which is very unbalanced. Amino acid metabolism is thus crucial in aphids, and we describe a novel investigation method based on the use of ¹⁴C-labeled amino acids added in an artificial diet. A metabolism cage for aphids was constructed, allowing for the collection and analysis of the radioactivity incorporated into the aphid body, expired as CO₂, and rejected in the honeydew and exuviae. This method was applied to the study of the metabolism of eight energetic amino acids (aspartate, glutamate, glutamine, glycine, serine, alanine, proline, and threonine) in the pea aphid, Acyrthosiphon pisum. All these amino acids except threonine were subject to substantial catabolism as measured by high ¹⁴CO₂ production. The highest turnover was displayed by aspartate, with 60% of its carbons expired as CO₂. For the first time in an aphid, we directly demonstrated the synthesis of three essential amino acids (threonine, isoleucine, and lysine) from carbons of common amino acids. The synthesis of these three compounds was only observed from amino acids that were previously converted into glutamate. This conversion was important for aspartate, and lower for alanine and proline. To explain the quantitative results of interconversion between amino acids, we propose a compartmentation model with the intervention of bacterial endosymbiotes for the synthesis of essential amino acids and with glutamate as the only amino acid supplied by the insect to the symbiotes. Moreover, proline exhibited partial conversion into arginine, and it is suggested that proline is probably indirectly involved in excretory nitrogen metabolism. © 1995 Wiley-Liss, Inc.     

Loss of organic acids, amino acids, k, and cl from barley roots treated anaerobically and with metabolic inhibitors

Excised roots of barley (Hordeum vulgare, var. Campana) lost organic acids, amino acids, K⁺, and Cl⁻ within 15 minutes after initiation of anaerobic treatment or treatment with NaCN and 2,4-dinitrophenol. Initial loss of organic acids when roots were placed under N₂ is attributed to a decarboxylation reaction, possibly catalyzed by phosphoenolpyruvate carboxykinase. Organic and amino acids began to leak from the roots to the bathing medium after 1 to 2 hours under N₂, indicating injury to cell membranes. During the first hour of anaerobic treatment, K⁺ loss from low-salt roots was equivalent to organic acid loss. Potassium loss from roots containing high levels of KCl was approximately equal to organic acid plus amino acid loss; and Cl⁻ loss was approximately equal to amino acid loss. It is postulated that, within cells, organic acids may electrostatically bind an equivalent quantity of cations and that amino acids may bind an equivalent quantity of both cations and anions.     

CO2 dark fixation in the halophytic species mesembryanthemum crystallinum

The time course of ¹⁴CO₂ dark fixation was studied in leaves of the facultatively halophytic plant species Mesembryanthemum crystallinum cultivated with and without 400 mM NaCl in the nutrient medium. It is generally known from the literature that plants grown under saline conditions incorporate ¹⁴C predominately into amino acids. By contrast in leaves of M. crystallinum grown on NaCl and exposed to ¹⁴CO₂ in the dark, relatively more radioactivity is incorporated in the organic acids (especially malate) than in amino acids. The data obtained are discussed in relation to the NaCl induced Crassulacean acid metabolism in M. crystallinum reported earlier.     

Decreases in Amino Acid and Acetylcholine Metabolism During Hypoxia

Abstract: Hypoxia impairs brain function by incompletely defined mechanisms. Mild hypoxia, which impairs memory and judgment, decreases acetylcholine (ACh) synthesis, but not the levels of ATP or the adenylate energy charge. However, the effects of mild hypoxia on the synthesis of the glucosederived amino acids [alanine, aspartate, γ-amino butyric acid (GABA), glutamate, glutamine, and serine] have not been characterized. Thus, we examined the incorporation of [U-¹⁴C]glucose into these amino acids and ACh during anemic hypoxia (injection of NaNO₂), hypoxic hypoxia (15 or 10% O₂), and hypoxic hypoxia plus hypercarbia (15 or 10% O₂ with 5% CO₂). In general, the synthesis of the amino acids and of ACh declined in parallel with each type of hypoxia we studied. For example, anemic hypoxia (75 mg/kg of NaNO₂) decreased the incorporation of [U-¹⁴C]glucose into the amino acids and into ACh similarly. [Percent inhibition: ACh (57.4), alanine (34.4), aspartate (49.2), GABA (61.9). glutamine (59.2), glutamate (51.0), and serine (36.7)]. A comparison of several levels (37.5, 75, 150, 225 mg/kg of NaNO₂) of anemic hypoxia showed a parallel decrease in the flux of glucose into ACh and into the amino acids whose synthesis depends on mitochondrial oxidation: GABA (r= 0.98), glutamate (r= 0.99), aspartate (r= 0.96), and glutamine (r= 0.97). The synthesis of the amino acids not dependent on mitochondrial oxidation did not correlate as well with changes in ACh metabolism: serine (r= 0.68) and alanine (r= 0.76). The decreases in glucose incorporation into ACh and into the amino acids with hypoxic hypoxia (15% or 10% O₂) or hypoxic hypoxia with 5% CO₂ were very similar to those with the two lowest levels of anemic hypoxia. Thus, any explanation of the brain's sensitivity to a decrease in oxygen availability must include the alterations in the metabolism of the amino acid neurotransmitters as well as ACh.     

S‐deficiency responsive accumulation of amino acids is mainly due to hydrolysis of the previously synthesized proteins – not to de novo synthesis in Brassica napus

To characterize the mechanisms of amino acid accumulation under sulphur (S)‐deficiency and its physiological significance in Brassica napus, stable isotopes ¹⁵N and ³⁴S were employed. The plants were exposed for 9 days to S‐deficient conditions (0.05 mM vs 1.5 mM sulphate). After 9 days of S‐deficiency, leaf‐osmotic potential and total chlorophyll content significantly decreased. S uptake decreased by 94%, whereas N uptake and biomass were not significantly changed. Using ¹⁵N and ³⁴S labelling, de novo synthesis of amino acids and proteins derived from newly absorbed NO₃^? and SO₄^2? and the content of N and S in the previously synthesized amino acids and proteins were quantified. At the whole plant level, S‐deficiency increased the pool of amino acids but resulted in strong decrease of incorporation of newly absorbed NO₃^? and SO₄^2? into amino acids by 22.2 and 76.6%, respectively, compared to the controls. Total amount of N and S incorporated into proteins also decreased by 28.8 and 62.1%, respectively. The levels of ¹⁴N‐ and ³²S‐proteins (previously synthesized proteins) strongly decreased, mainly in mature leaves. The data thus indicate that amino acid accumulation under short‐term S‐deficiency results from the degradation of previously synthesized proteins rather than from de novo synthesis.     

Equilibration of H labeling between body water and free amino acids: Enabling studies of proteome synthesis

Protein synthesis can be estimated by measuring the incorporation of a labeled amino acid into a proteolytic peptide. Although prelabeled amino acids are typically administered, recent studies have tested ²H₂O; the assumption is that there is rapid equilibration of ²H (in body water) with the carbon-bound hydrogens of amino acids before those amino acids are incorporated into a protein(s). We have determined the temporal changes in ²H labeling of body water and amino acids which should build confidence in ²H₂O-based studies of protein synthesis when one aims to measure the ²H labeling of proteolytic peptides.     

Effect of Abscisic Acid on the Activities of Photosynthetic Enzymes and 14CO2 Fixation Products in Leaves of Pennisetum typhoides Seedlings

Abscisic acid inhibited the rate of ¹⁴CO₂ fixation in leaves of Pennisetum typhoides (Burm. f.) Stapf & Hubbard seedlings, but increased the activities of phosphoenol-pyruvate-carboxylase and malic enzyme. The leaves of the seedlings grown in the presence of abscisic acid incorporated, in comparison to the control, more radioactivity in the fraction of organic acids, but less radioactivity was recorded in the amino acid fraction. On the other hand, gibberellic acid which also inhibits photosynthetic ¹⁴CO₂ assimilation and decreases the activities of photosynthetic enzymes, favours greater incorporation in alanine, and reduces that in malate. It is deduced that bio-regulants can greatly influence the flow of ¹⁴C into individual photosynthetic products. As in growth, abscisic and gibberellic acids in combination tended to antagonize each other in their effects on enzyme activity as well as in incorporation of ¹⁴CO₂ into photosynthetic products.     

Regulation of nitrate uptake by amino acids in maize cell suspension culture and intact roots

The effect of amino acids on nitrate transport was studied in Zea mays cell suspension cultures and in Zea mays excised roots. The inclusion of aspartic acid, arginine, glutamine and glycine (15mM total amino acids) in a complete cell-culture media containing 1.0 mM NO₃ ^- strongly inhibited nitrate uptake and the induction of accelerated uptake rates. The nitrate uptake rate increased sharply once solution amino acid levels fell below detection limits. Glutamine alone inhibited induction in the cell suspension culture. Maize seedlings germinated and grown for 7 days in a 15 mM mixture of amino acids also had lower nitrate uptake rates than seedlings grown in 0.5 mM Ca(NO₃)₂ or 1 mM CaCl₂. As amino acids are the end product of nitrate assimilation, the results suggest an end-product feed-back mechanism for the regulation of nitrate uptake.     

Physiological changes in white lupin associated with variation in root-zone CO2 concentration and cluster-root P mobilization

White lupin (Lupinus albus L.) mobilizes insoluble soil phosphorus through exudation of organic acids from ‘cluster’ roots. Organic acid synthesis requires anaplerotic carbon derived from dark CO₂ fixation involving PEP-carboxylase. We tested the hypothesis that variation in root-zone CO₂ concentration would influence organic acid synthesis and thus P mobilization. Root-zone CO₂ concentrations and soil FePO₄ concentrations supplied to sand-grown white lupin (cv. Kiev Mutant) were varied. More biomass accumulated in plants supplied with 360 µL L⁻¹ CO₂ to the root-zone, compared with those aerated with either 100 or 6000 µL L⁻¹ CO₂. Increased FePO₄ in the sand resulted in greater leaf P concentrations, but root-zone [CO₂] did not influence leaf P concentration. Suppression of cluster-root development in plants supplied with 100 µL L⁻¹ root-zone CO₂ was correlated with increased leaf [P]. However, at both 360 and 6000 µL L⁻¹ CO₂, cluster-root development was suppressed only at the highest leaf P concentration. Phloem sap [P] was significantly increased by greater [FePO₄] in the sand, but was reduced with increased root-zone [CO₂], and this may have triggered increased cluster-root initiation. Succinate was the major organic acid (carboxylate) in the phloem sap (minor components included malate, citrate, fumarate) and was increased at greater [FePO₄], suggesting that this shoot-derived carboxylate might provide an important source of organic acids for root metabolism. Since cluster root development was inhibited by increasing concentrations of FePO₄ in the sand, it is possible that succinate was utilized for the functioning of the root-nodules.     

KINETICS OF COMPETITIVE INHIBITION OF NEUTRAL AMINO ACID TRANSPORT ACROSS THE BLOOD-BRAIN BARRIER

The transport of tryptophan across the blood-brain barrier is used as a specific example of a general approach by which rates of amino acid influx into brain may be predicted from existing concentrations of amino acids in plasma. The kinetics of inhibition of [¹⁴C]tryptophan transport by four natural neutral amino acids (phenylalanine, leucine, methionine, and valine) and one synthetic amino acid (α-methyl tyrosine) is studied with a tissue-sampling, single injection technique in the barbiturate-anesthetized rat. The equality of the K₁ (determined from cross-inhibition studies) and the K_m (determined from auto-inhibition data) for neutral amino acid transport indicate that these amino acids compete for a single transport site in accordance with the kinetics of competitive inhibition. Based on equations derived for competitive inhibition, apparent K_m values are computed for the essential neutral amino acids from known data on amino acid transport K_m and plasma concentrations. The apparent K_m values make possible predictions of the in vivo rates of amino acid influx into brain based on given plasma amino acid concentrations. Finally, a method is presented for determining transport constants from saturation data obtained with single injection techniques.     

Die Aufnahme von 14CO2 und die Verteilung des 14C auf freie Aminosäuren und auf Proteinaminosäuren im Hellrot und im Blaulicht. [Objekt: Farnvorkeime von Dryopteris filix-mas (L.) Schott]

Summary Morphogenesis and metabolism of the early gametophytes (= sporelings) of the common male fern are controlled by light. The normal two-dimensional development of the gametophytes in white or blue light is correlated with an increase in protein content; inred light alone, on the other hand, the sporelings remain filamentous, and the protein content is markedly lower (cf. Mohr, 1965). The problem has been whether blue light increases the rate of protein synthesis or decreases the rate of protein degradation. This problem was solved in the present paper by the use of ¹⁴CO₂. Blue light promotes specifically the rate of protein synthesis as indicated by the increase of ¹⁴C incorporation into protein-bound amino acids under blue light as compared with red light.Using ¹⁴CO₂ we have analyzed the kinetics of free amino acid synthesis (Fig. 4) and protein synthesis (Fig. 5) under steady state conditions of photosynthetic CO₂ incorporation in blue or red light (Fig. 3). Under our conditions the rate of photosynthesis is about 1.5 times higher under blue light than under red light (Fig. 3, Table 1).The facts that the total pool sizes of the free amino acids are smaller in blue than in red light (v. Deimling and Mohr, 1967; Table 2) and that, on the other hand, the ¹⁴C-contents of the thoroughly labelled amino acid pools are virtually identical in blue and red (Table 3) indicate (a) that the pool sizes of these labelled amino acids may be equal in both light qualities and (b) that there is a compartmentation of free amino acid pools in the fern sporeling. This problem will be dealt with more in detail in a forthcoming paper on the behaviour of alanine in the fern sporeling (Payer, 1969).Protein synthesis is obviously much stronger under blue light than under red light. The detailed kinetics (Fig. 5b) indicate the involvement of two sorts of proteins: a relatively small part with high turnover which is rapidly labelled with a small but significant difference in red and blue, and a larger part with a slower turnover, the synthesis of which is strongly favored by blue light. — The first sort could be enzyme protein; the latter sort might be structural protein of the chloroplasts. These organelles increase dramaticly in size under the influence of blue light (Bergfeld, 1963). The amino acid composition of the protein, however, does not show any qualitative difference in gametophytes grown in blue or red light (v. Deimling and Mohr, 1967, Table 4).

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Die Aufnahme von ¹⁴CO₂ und die Verteilung des ¹⁴C auf freie Aminosäuren und auf Proteinaminosäuren im Hellrot und im Blaulicht. [Objekt: Farnvorkeime von Dryopteris filix-mas (L.) Schott]

     

Relationship between leaf development and primary photosynthetic products in the C4 plant Portulaca oleracea L.

Summary The photosynthetic products of Portulaca oleracea differ greatly depending on leaf age and length of exposure to ¹⁴CO₂. Mature leaves of P. oleracea fix ¹⁴CO₂ primarily into organic and amino acids during a 10-s exposure period. Less than 2% of the ¹⁴CO₂ fixed appears in phosphorylated compounds. In contrast, incorporation into amino acids can account for over 60% of the total ¹⁴CO₂ fixed by young leaves in an equal time period, and incorporation into alanine alone can account for up to one half of this amount. Senescent leaves display a quantitative shift of primary products toward phosphorylated compounds with a concomitant reduction of the label residing in malate and asparate. About 8 times more phosphoglyceric acid is produced in senescent leaves than in mature leaves. The aspartate/ malate ratio is not constant and depends on the length of time the leaves are exposed to ¹⁴CO₂ and the age of the leaves under study. It appears as if the stage of leaf development is one of the most important factors determining the operation of a particular enzyme system in C₄ plants.     

Developmental control of H+/amino acid permease gene expression during seed development of Arabidopsis

Long distance transport of amino acids is mediated by several families of differentially expressed amino acid transporters. The two genes AAP1 and AAP2 encode broad specificity H⁺-amino acid co-transporters and are expressed to high levels in siliques of Arabidopsis, indicating a potential role in supplying the seeds with organic nitrogen. The expression of both genes is developmentally controlled and is strongly induced in siliques at heart stage of embryogenesis, shortly before induction of storage protein genes. Histochemical analysis of transgenic plants expressing promoter-GUS fusions shows that the genes have non-overlapping expression patterns in siliques. AAP1 is expressed in the endosperm and the cotyledons whereas AAP2 is expressed in the vascular strands of siliques and in funiculi. The endosperm expression of AAP1 during early stages of seed development indicates that the endosperm serves as a transient storage tissue for organic nitrogen. Amino acids are transported in both xylem and phloem but during seed filling are imported only via the phloem. AAP2, which is expressed in the phloem of stems and in the veins supplying seeds, may function in uptake of amino acids assimilated in the green silique tissue, in the retrieval of amino acids leaking passively out of the phloem and in xylem-to-phloem transfer along the path. The promoters provide excellent tools to study developmental, hormonal and metabolic control of nitrogen nutrition during development and may help to manipulate the timing and composition of amino acid import into seeds.