Carrier-mediated uptake of glyphosate in broad bean (Vicia faba) via a phosphate transporter

The possibility that the herbicide glyphosate (N-phosphonomethylglycine) may be taken up in plant cells via a phosphate transporter of the plasma membrane was investigated using protoplasts of broad bean leaves ( Vicia faba L.). Phosphonoformic acid, a powerful inhibitor of phosphate transport in animal cells, was first demonstrated to be a competitive inhibitor of phosphate uptake inbroad bean protoplasts. Glyphosate was able to inhibit phosphate uptake into the protoplasts, and to protect partially the phosphate transporter from inhibition by phosphonoformic acid. Concentration dependence studies showed that glyphosate uptake exhibited a saturable phase at low glyphosate concentrations (0. 5 to 3 μ M ), superimposed by a linear uptake at higher concentrations (up to 100 μ M ). Inhibition of glyphosate uptake by para -chloromercuribenzene sulphonic acid, sodium azide and carbonyl-cyanide- m -chlorophenylhydrazone was much stronger at 1 than at 100 μ M glyphosate. Kinetics indicated that the saturable component of glyphosate transport was competitively inhibited by either phosphate or phosphonoformic acid. It is concluded that glyphosate can be absorbed via a phosphate transporter of the plasma membrane     

Transporters in Arabidopsis roots mediating uptake of amino acids at naturally occurring concentrations

Recent studies of Arabidopsis have identified several transporters as being important for amino acid uptake. We used Arabidopsis plants with altered expression of lysine histidine transporter 1 (LHT1), amino acid permease 1 (AAP1) and amino acid permease 5 (AAP5) with the aim of disentangling the roles of each transporter in the uptake of different amino acids at naturally occurring concentrations (2-50 μM). LHT1 mutants displayed reduced uptake rates of L-Gln, L-Ala, L-Glu and L-Asp but not of L-Arg or L-Lys, while AAP5 mutants were affected in the uptake of L-Arg and L-Lys only. Double mutants (lht1aap5) exhibited reduced uptake of all tested amino acids. In the concentration range tested, AAP1 mutants did not display altered uptake rates for any of the studied amino acids. Expression analysis of amino acid transporter genes with important root functions revealed no major differences in the individual mutants other than for genes targeted for mutation. We conclude that LHT1 and AAP5, but not AAP1, are crucial for amino acid uptake at concentrations typically found in soils. LHT1 and AAP5 displayed complementary affinity spectra, and no redundancy with respect to gene expression was found between the two transporters, suggesting these two transporters have separate roles in amino acid uptake.     

Characteristics of amino acid uptake in barley

Plants have the ability to take up organic nitrogen (N) but this has not been thoroughly studied in agricultural plants. A critical question is whether agricultural plants can acquire amino acids in a soil ecosystem. The aim of this study was to characterize amino acid uptake capacity in barley (Hordeum vulgare L.) from a mixture of amino acids at concentrations relevant to field conditions. Amino acids in soil solution under barley were collected in microlysimeters. The recorded amino acid composition, 0–8.2 μM of l-Serine, l-Glutamic acid, Glycine, l-Arginine and l-Alanine, was then used as a template for uptake studies in hydroponically grown barley plants. Amino acid uptake during 2 h was studied at initial concentrations of 2–25 μM amino acids and recorded as amino acid disappearance from the incubation solution, analysed with HPLC. The uptake was verified in control experiments using several other techniques. Uptake of all five amino acids occurred at 2 μM and below. The concentration dependency of the uptake rate could be described by Michaelis–Menten kinetics. The affinity constant (K _m) was in the range 19.6–33.2 μM. These K _m values are comparable to reported values for soil micro-organisms.     

Identification,functional characterization and expression of a LAT type amino acid transporter from the mosquito Aedes aegypti

We isolated two cDNAs from the mosquito Aedes aegypti, an L-amino acid transporter (AeaLAT) and a CD98 heavy chain (AeaCD98hc). Expression of AeaCD98hc or AeaLAT alone in Xenopus oocyte did not induce amino acid transport activity. However, co-expression of AeaCD98hc and AeaLAT, which are postulated to form a heterodimer protein linked through a disulfide bond, showed significant increase in amino acid transport activity. This heterodimeric protein showed uptake specificity for large neutral and basic amino acids. Small acidic neutral amino acids were poor substrates for this transporter. Neutral amino acid (leucine) uptake activity was partially Na+ dependent, because leucine uptake was approximately 44% lower in the absence of Na+ than in its presence. However, basic amino acid (lysine) uptake activity was completely Na+ independent at pH of 7.4. Extracellular amino acid concentration could be the main factor that determined amino acid transport. These results suggest the heteromeric protein is likely a uniporter mediating diffusion of amino acids in the absence of ions. The AeaLAT showed high level expression in the gastric caeca, Malpighian tubules and hindgut of larvae. In caeca and hindgut expression was in the apical cell membrane. However, in Malpighian tubules and in midgut, the latter showing low level expression, the transporter was detected in the basolateral membrane. This expression profile supports the conclusion that this AeaLAT is a nutrient amino acid transporter.     

Cationic amino acid transporter 1-mediated l-arginine transport at the inner blood–retinal barrier

The purpose of this study was to identify the transporter mediating l -arginine transport at the inner blood–retinal barrier (BRB). The apparent uptake clearance of [³H] l -arginine into the rat retina was found to be 118 μL/(min·g retina), supporting a carrier-mediated influx transport of l -arginine at the BRB. [³H] l -Arginine uptake by a conditionally immortalized rat retinal capillary endothelial cell line (TR-iBRB2 cells), used as an in vitro model of the inner BRB, was primarily an Na⁺-independent and saturable process with Michaelis-Menten constants of 11.2 μM and 530 μM. This process was inhibited by rat cationic amino acid transporter (CAT) 1-specific small interfering RNA as well as substrates of CATs, l -arginine, l -lysine, and l -ornithine. The expression of cationic amino acid transporter (CAT) 1 mRNA was 25.9- and 796-fold greater than that of CAT3 in TR-iBRB2 and magnetically isolated rat retinal vascular endothelial cells, respectively. The expression of CAT1 protein was detected in TR-iBRB2 cells and immunostaining of CAT1 was observed along the rat retinal capillaries. In conclusion, CAT1 is localized in retinal capillary endothelial cells and at least in part mediates l -arginine transport at the inner BRB. This process seems to be closely involved in visual functions by supplying precursors of biologically important molecules like nitric oxide in the neural retina.     

Functional Characterization of the Amanita muscaria Monosaccharide Transporter, AmMst1

Abstract: Fungal carbohydrate nutrition is an important aspect of ectomycorrhizal symbiosis. At the plant/fungus interface, fungal and root cortical cells compete for monosaccharides, generated from plant-derived sucrose. Therefore, the kinetic properties of the monosaccharide uptake systems are decisive for the monosaccharide yield of each partner. For the functional characterization of a hexose transporter (AmMst1) of the ectomycorrhizal fungus Amanita muscaria, the entire cDNA was expressed in a Saccharomyces cerevisiae strain unable to take up hexoses. Uptake experiments with ¹⁴C-labelled monosaccharides resulted in K_M values of 0.46 mM for glucose and 4.20 mM for fructose, revealing a strong preference of AmMst1 for glucose as substrate. Glucose uptake by AmMst1 was strongly favoured even in the presence of a large excess of fructose. Comparable affinities of AmMst1 for glucose, 3-O-methyl glucose and mannose were obtained. In contrast, AmMst1 imported galactose with a much lower efficiency, revealing that this transporter distinguishes pyranoses by steric hindrance at the C-4 position. While yeast contains numerous hexose transporter genes, the AmMst1 gene seems to be the main, if not the only, hexose transporter that is expressed in A. muscaria, as concluded from the comparison of hexose import properties of A. muscaria protoplasts and AmMst1 expressed in yeast.     

Uptake of amino acids by the aquatic resurrection plant Chamaegigas intrepidus and its implication for N nutrition

Chamaegigas intrepidus is a poikilohydric aquatic plant that lives in rock pools on granitic outcrops in Central Namibia. The pools are filled intermittently during the summer rains, and the plants may pass through up 20 rehydration/dehydration cycles during a single wet season. Rehydrated plants also have to cope with substantial diurnal fluctuations in the pH and extreme nutrient deficiency. Ammonium concentrations are normally around 30 μM. Additional nitrogen sources are amino acids. Total free amino acids are up to 15 μM with glycine and serine as the predominant amino acids. Experiments on uptake of radiolabelled amino acids into roots of C. intrepidus showed high␣affinity (K _M= 16 μM) and low-affinity (K _M= 159 μM) uptake systems. The K _M of the high-affinity system is well in accordance with the free amino acid concentration found in the water of the pools. We conclude that amino acids, predominantly glycine and serine, can be utilised by C. intrepidus in its natural habitat. Since glycine uptake showed a strong reduction at pH 10, nitrogen uptake from glycine or serine should occur mainly in the morning when the pH of the pool water is slightly acid. Further experiments with ¹⁵N-labelled ammonium in combination with non-labelled glycine demonstrated high ¹⁵N values in plant tissues. Under experimental conditions C. intrepidus preferred ammonium as a nitrogen source. The implication of amino acids for nitrogen nutrition of C. intrepidus may depend on the relation of inorganic and organic nitrogen available in the pool water and the preferential utilisation of one or the other nitrogen source may change during the day corresponding with pH changes in the water. Received: 28 January 1998 / Accepted: 24 July 1998     

Triiodothyronine Is a High-Affinity Inhibitor of Amino Acid Transport System L1 in Cultured Astrocytes

Abstract: The relationship between the transport of thyroid hormones and that of amino acids was examined by measuring the uptake of amino acids that are characteristic substrates of systems L, A, and N, and the effect of 3,3',5-triiodo-L-thyronine (T₃) on this uptake, in cultured astrocytes. Tryptophan and leucine uptakes were rapid, Na⁺-independent, and efficiently inhibited by T₃ (half-inhibition at ∼ 2 μ M ). Two Na⁺-independent L-like systems (L₁ and L₂), common to leucine and aromatic amino acids, were characterized kinetically. System L₂ had a low affinity for leucine and tryptophan ( K _m= 0.3–0.9 m M ). The high-affinity system L₁ ( K _m∼ 10 μ M for both amino acids) was competitively inhibited by T₃ with a K _i of 2–3 μ M (close to the T₃ transport K _m). Several T₃ analogues inhibited system L₁ and the T₃ transport system similarly. Glutamine uptake and α-(methylamino)isobutyric acid uptake were, respectively, two and 200 times lower than tryptophan and leucine uptakes. T₃ had little effect on the uptakes of glutamine and α-(methylamino)isobutyric acid. The results indicate that the T₃ transport system and system L₁ are related.     

Basolateral aromatic amino acid transporter TAT1 (Slc16a10) functions as an efflux pathway

Basolateral efflux is a necessary step in transepithelial (re)absorption of amino acids from small intestine and kidney proximal tubule. The best characterized basolateral amino acid transporters are y+LAT1-4F2hc and LAT2-4F2hc that function as obligatory exchangers and thus, do not contribute to net amino acid (re)absorption. The aromatic amino acid transporter TAT1 was shown previously to localize basolaterally in rat's small intestine and to mediate the efflux of L-Trp in the absence of exchange substrate, upon expression in Xenopus oocytes. We compared here the amino acid influx and efflux via mouse TAT1 in Xenopus oocytes. The results show that mTAT1 functions as facilitated diffusion pathway for aromatic amino acids and that its properties are symmetrical in terms of selectivity and apparent affinity. We show by real-time RT-PCR that its mRNA is highly expressed in mouse small intestine mucosa, kidney, liver, and skeletal muscle as well as present in all other tested tissues. We show that mTAT1 is not N-glycosylated and that it localizes to the mouse kidney proximal tubule. This expression is characterized by an axial gradient similar to that of the luminal neutral amino acid transporter B0AT1 and shows the same basolateral localization as 4F2hc. mTAT1 also localizes to the basolateral membrane of small intestine enterocytes and to the sinusoidal side of perivenous hepatocytes. In summary, we show that TAT1 is a basolateral epithelial transporter and that it can function as a net efflux pathway for aromatic amino acids. We propose that it, thereby, may supply parallel exchangers with recycling uptake substrates that could drive the efflux of other amino acids.     

Cloning,Expression, and Functional Characterization of Secondary Amino Acid Transporters of Lactococcus lactis

Fourteen genes encoding putative secondary amino acid transporters were identified in the genomes of Lactococcus lactis subsp. cremoris strains MG1363 and SK11 and L. lactis subsp. lactis strains IL1403 and KF147, 12 of which were common to all four strains. Amino acid uptake in L. lactis cells overexpressing the genes revealed transporters specific for histidine, lysine, arginine, agmatine, putrescine, aromatic amino acids, acidic amino acids, serine, and branched-chain amino acids. Substrate specificities were demonstrated by inhibition profiles determined in the presence of excesses of the other amino acids. Four knockout mutants, lacking the lysine transporter LysP, the histidine transporter HisP (formerly LysQ), the acidic amino acid transporter AcaP (YlcA), or the aromatic amino acid transporter FywP (YsjA), were constructed. The LysP, HisP, and FywP deletion mutants showed drastically decreased rates of uptake of the corresponding substrates at low concentrations. The same was observed for the AcaP mutant with aspartate but not with glutamate. In rich M17 medium, the deletion of none of the transporters affected growth. In contrast, the deletion of the HisP, AcaP, and FywP transporters did affect growth in a defined medium with free amino acids as the sole amino acid source. HisP was essential at low histidine concentrations, and AcaP was essential in the absence of glutamine. FywP appeared to play a role in retaining intracellularly synthesized aromatic amino acids when these were not added to the medium. Finally, HisP, AcaP, and FywP did not play a role in the excretion of accumulated histidine, glutamate, or phenylalanine, respectively, indicating the involvement of other transporters.     

Free amino acids production by ectomycorrhizal fungi of pine (Pinus sylvestris L.)

Studies on free amino acids production by five species of ectomycorrhizal fungi (Amanita muscaria, Suillus granulatus, Suillus luteus, Suillus bovinus and Rhizopogon luteolus) show that all the fungi produced mainly: glutamic acid, leucine, lysine, ornithine, arginine and an unidentified ninhydrin-positive compound X3. Both the quality and quantity of amino acids released was different in the fungal species studied. The predominant amino acids in post-culture liquids in general did not exceed 1.5 micrograms/mg dry mass.     

丛枝菌根真菌萌发孢子利用不同氮素及外源葡萄糖对其代谢的影响

对各种含氮基质、葡萄糖和(或)根浸出液中培养的丛枝菌根真菌Glomus intraradices孢子,在萌发过程中对不同氮素的利用及其氨基酸的生物合成进行了研究.用稳定同位素标记及质谱仪来分析不同氮素的利用和氨基酸的生物合成.以高效液相色谱测量氨基酸的浓度.在缺少外源氮素的情况下,丛枝菌根真菌孢子萌发时可以利用内部储存的含氮化合物生物合成游离氨基酸.其中,丝氨酸和甘氨酸是大量合成的氨基酸.合成的氨基酸浓度在2周内随着萌发时间的增加而增加.在有可利用的外源无机氮(铵盐、硝酸盐和尿素)和有机氮(氨基酸)时,铵盐和尿素比硝酸盐更容易被AM真菌萌发孢子利用,而其利用氨基酸中的氮比无机氮源慢的多.孢子吸收同化外源无机氮,且将其整合到游离氨基酸中,这些新生氨基酸浓度比无外源氮添加时要高得多.在无葡萄糖添加的硝酸盐培养液中,AM真菌孢子中积累大量天冬酰胺.然而,在含有葡萄糖的培养液中,萌发孢子因葡萄糖的吸收促进了对外源氮的吸收,产生的游离氨基酸是无葡萄糖时的5倍,并且发现精氨酸转为含量最多的游离氨基酸.并且,从外源氮吸收同化的氮可以储存于精氨酸中,随之,精氨酸被整合到AM真菌孢子储存的蛋白质中.此外,根浸出原液在AM真菌孢子萌发2周后对氮的吸收作用不明显.     

Arabidopsis LHT1 is a high-affinity transporter for cellular amino acid uptake in both root epidermis and leaf mesophyll

Amino acid transport in plants is mediated by at least two large families of plasma membrane transporters. Arabidopsis thaliana, a nonmycorrhizal species, is able to grow on media containing amino acids as the sole nitrogen source. Arabidopsis amino acid permease (AAP) subfamily genes are preferentially expressed in the vascular tissue, suggesting roles in long-distance transport between organs. We show that the broad-specificity, high-affinity amino acid transporter LYSINE HISTIDINE TRANSPORTER1 (LHT1), an AAP homolog, is expressed in both the rhizodermis and mesophyll of Arabidopsis. Seedlings deficient in LHT1 cannot use Glu or Asp as sole nitrogen sources because of the severe inhibition of amino acid uptake from the medium, and uptake of amino acids into mesophyll protoplasts is inhibited. Interestingly, lht1 mutants, which show growth defects on fertilized soil, can be rescued when LHT1 is reexpressed in green tissue. These findings are consistent with two major LHT1 functions: uptake in roots and supply of leaf mesophyll with xylem-derived amino acids. The capacity for amino acid uptake, and thus nitrogen use efficiency under limited inorganic N supply, is increased severalfold by LHT1 overexpression. These results suggest that LHT1 overexpression may improve the N efficiency of plant growth under limiting nitrogen, and the mutant analyses may enhance our understanding of N cycling in plants.     

Kinetic characterization of sulphur-containing excitatory amino acid uptake in primary cultures of neurons and astrocytes

The uptake of the neuroactive sulphur amino acids -cysteine sulphinate, -cysteate, -homocysteine sulphinate and -homocysteate was investigated in astrocytes cultured from the prefrontal cortex; in neurons, cultured from cerebral cortex; and, in granule cells, cultured from cerebellum. It was shown that each amino acid acted as a substrate for a plasma membrane transporter in both neurons and astrocytes. Astrocytes and neurons exhibited a high-affinity uptake for -cysteine sulphinate and -cysteate with K_m values ranging from 14–100 μM, and a low-affinity uptake for -homocysteine sulphinate and -homocysteate, with K_m values ranging from 225–1210 μM. The uptake of all transmitter candidates studied was partially sodium-dependent. This sodium-dependency was most evident at low (< 100 μM) concentrations of each substrate. The apparent uptake measured in the absence of sodium was included as a component in corrections made for non-saturable influx. With the exception of -cysteine sulphinate, uptake of each sulphur amino acid was greatest in astrocytes, with V_max values ranging between 15–32 nmol min⁻¹ mg⁻¹ cell protein. Moreover, the uptake of each sulphur amino acid in cerebellar granule cells (V_max values ranging between 10–25 nmol min⁻¹ mg⁻¹ cell protein) was consistently greater than that in cerebral cortex neurons (V_max values ranging between 1.5–6 nmol min⁻¹ mg⁻¹ cell protein).     

Kinetics of amino acid uptake by ectomycorrhizal roots

It is well established that ectomycorrhizal fungi can use amino acids as nitrogen and carbon sources, but data on the kinetic properties of amino acid uptake systems of ectomycorrhizal systems are scarce. Using ¹⁴C-labelled compounds we have determined the kinetics of uptake of amino acids by excised ectomycorrhizal roots for a range of distinct mycorrhizal types from three tree species, beech, spruce, and pine. All mycorrhizal types examined took up amino acids via high-affinity transport systems ( K _M values ranging from 19 to 233 mmol m^–3). A comparative analysis of kinetic parameters for uptake of amino acids and the ammonium analogue methylammonium showed that ectomycorrhizal roots have similar or even higher affinities (lower K _M values) for the amino acids, indicating that absorption of these organic forms of nitrogen (N) can contribute significantly to total N uptake by ectomycorrhizal plants. Analysis of amino acid uptake by ectomycorrhizal roots collected along a European north/south gradient of increasing mineral N pollution from northern Sweden to south Germany revealed no obvious trend in the uptake capabilities for amino acids by ectomycorrhizal roots in relation to the location of the sampling site on this gradient. Rather, the fungal species forming a particular morphotype was the factor determining uptake kinetics. It can therefore be deduced that the species composition of the fungal community will contribute significantly to the functional diversity of a population of mycorrhizal roots.     

Mutational analysis of histidine residues in the human proton-coupled amino acid transporter PAT1

The proton-coupled amino acid transporter 1 (PAT1) represents a major route by which small neutral amino acids are absorbed after intestinal protein digestion. The system also serves as a novel route for oral drug delivery. Having shown that H+ affects affinity constants but not maximal velocity of transport, we investigated which histidine residues are obligatory for PAT1 function. Three histidine residues are conserved among the H+-coupled amino acid transporters PAT1 to 4 from different animal species. We individually mutated each of these histidine residues and compared the catalytic function of the mutants with that of the wild type transporter after expression in HRPE cells. His-55 was found to be essential for the catalytic activity of hPAT1 because the corresponding mutants H55A, H55N and H55E had no detectable l-proline transport activity. His-93 and His-135 are less important for transport function since H93N and H135N mutations did not impair transport function. The loss of transport function of His-55 mutants was not due to alterations in protein expression as shown both by cell surface biotinylation immunoblot analyses and by confocal microscopy. We conclude that His-55 might be responsible for binding and translocation of H+ in the course of cellular amino acid uptake by PAT1.     

Solute transport into healthy and powdery mildew-infected leaves of pea and uptake by powdery mildew mycelium

The transport of sugars and amino acids into the mycelium of Erysiphe pisi DC. was investigated using two different systems, intact leaf discs and mycelial suspensions. Of the sugars tested, glucose was preferentially taken up by both uninfected and mildew-infected leaf discs, whereas glutamine was taken up by both tissues at a higher rate than lysine or aspartic acid. Leaf discs from infected tissue had a greater uptake capacity than those from healthy tissue for both sugars and amino acids. The uptake of glucose was inhibited more markedly than that of sucrose and fructose by 10 μ m carbonyl cyanide m -chlorophenylhydrazone (CCCP), 1 m m N -ethylmaleimide (NEM), 1 m m diethyl pyrocarbonate (DEPC) and 1 m m phenylglyoxal, whereas 1 m m PCMBS ( p -chloro-mercuribenzenesulphonic acid) inhibited sucrose uptake to the greatest extent. Uptake of glutamine, lysine and aspartic acid was inhibited similarly by CCCP (80%), NEM (20%), DEPC (70%) and PCMBS (60%). Additionally, leaf discs were used to determine which solutes could be taken up from leaf tissue by the fungus. The uptake of sugars into the mycelium was greater than that of amino acids.
Suspensions of powdery mildew mycelium accumulated glucose at about three times the rate of sucrose or fructose, and the amino acid glutamine was taken up at three times the rate of lysine or aspartic acid. Spores separated from the suspension had a low uptake capacity.
When the reducing sugar concentration of leaf apoplastic fluid was estimated, leaves infected by powdery mildew had much higher amounts in the apoplast, whereas the activity of acid invertase also appeared to be higher in apoplastic fluids from infected leaves. When apoplastic fluid samples were run on SDS gels, an invertase antibody detected two bands in samples from infected tissues that were not found in the uninfected samples.     

Effects of Free Fatty Acids on Synaptosomal Amino Acid Uptake Systems

Abstract: The Na⁺-dependent synaptosomal uptakes of proline, aspartic acid, glutamic acid, and γ-aminobutyric acid were strongly inhibited by monounsaturated fatty acids. With oleic acid, half-maximal inhibition was observed at about 15 μM. The Na⁺-independent uptakes of leucine, phenylalanine, histidine, and valine were less sensitive to inhibition by the unsaturated fatty acids. In contrast, the uptakes of all of these amino acids were unaffected by saturated fatty acids. The inhibition of proline uptake (and that of the other Na⁺-dependent amino acids) by oleic acid was overcome by the addition of serum albumin and the data presented further indicate that the previously reported stimulation of proline uptake by albumin could be related to its fatty acid binding properties.     

Mutational analysis of histidine residues in the human proton-coupled amino acid transporter PAT1

The proton-coupled amino acid transporter 1 (PAT1) represents a major route by which small neutral amino acids are absorbed after intestinal protein digestion. The system also serves as a novel route for oral drug delivery. Having shown that H⁺ affects affinity constants but not maximal velocity of transport, we investigated which histidine residues are obligatory for PAT1 function. Three histidine residues are conserved among the H⁺-coupled amino acid transporters PAT1 to 4 from different animal species. We individually mutated each of these histidine residues and compared the catalytic function of the mutants with that of the wild type transporter after expression in HRPE cells. His-55 was found to be essential for the catalytic activity of hPAT1 because the corresponding mutants H55A, H55N and H55E had no detectable l-proline transport activity. His-93 and His-135 are less important for transport function since H93N and H135N mutations did not impair transport function. The loss of transport function of His-55 mutants was not due to alterations in protein expression as shown both by cell surface biotinylation immunoblot analyses and by confocal microscopy. We conclude that His-55 might be responsible for binding and translocation of H⁺ in the course of cellular amino acid uptake by PAT1.     

Uptake of intact amino acids by plants depends on soil amino acid concentrations

Studies in different ecosystems have shown that plants take up intact amino acids directly but little is known about the influence of free amino acid concentrations in the soil on this process. We investigated the effect of three different soil amino acid N concentrations (0.025, 0.13 and 2.5 μg N g^?1 soil) on direct uptake of four dual labelled (¹⁵N, ¹³C) amino acids (glycine, tyrosine, lysine, valine) in a greenhouse experiment using Anthoxantum odoratum as a model plant.Our results revealed that 8–45% of applied ¹⁵N was incorporated into plant root and shoot tissue 48 h after labelling. Additional ¹³C enrichment showed that 2–70% of this incorporated ¹⁵N was taken up as intact amino acid. Total ¹⁵N uptake and ¹⁵N uptake as intact amino acids were significantly affected by soil amino acid N concentrations and significantly differed between the four amino acids tested.We found a positive effect of soil amino acid concentrations on uptake of mineralized ¹⁵N relative to amino acid concentrations for all amino acids which was presumably due to higher diffusion rates of mineralized tracer to the root surface. However, intact amino acid uptake relative to amino acid concentrations as well as the proportion of total ¹⁵N taken up directly decreased with increasing soil amino acid N concentrations for all amino acids, irrespective of their microbial degradability. This effect is most likely controlled by the mineral N concentration in soil and perhaps in plants which inhibits direct amino acids uptake.Overall, we conclude that plant internal regulation of amino acid uptake controlled by mineral N is the main mechanism determining direct uptake of amino acids and thus a lower contribution of intact amino acid uptake to the plants N nutrition has to be expected for higher amino acid concentrations accompanied by mineralization in soil.