Sandra Jämtgård

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.

Plant nitrogen status and co-occurrence of organic and inorganic nitrogen sources influence root uptake by Scots pine seedlings

Insights into how the simultaneous presence of organic and inorganic nitrogen (N) forms influences root absorption will help elucidate the relative importance of these N forms for plant nutrition in the field as well as for nursery cultivation of seedlings. Uptake of the individual N forms arginine, ammonium (NH4(+)) and nitrate (NO3(-)) was studied in Scots pine (Pinus sylvestris (L.)) seedlings supplied as single N sources and additionally in mixtures of NO3(-) and NH4(+) or NO3(-) and arginine. Scots pine seedlings displayed a strong preference for NH4(+)-N and arginine-N as compared with NO3(-)-N. Thus, NO3(-) uptake was generally low and decreased in the presence of NH4(+) in the high-concentration range (500 µM N), but not in the presence of arginine. Moreover, uptake of NO3(-) and NH4(+) was lower in seedlings displaying a high internal N status as a result of high N pre-treatment, while arginine uptake was high in seedlings with a high internal N status when previously exposed to organic N. These findings may have practical implications for commercial cultivation of conifers.

Amino acid transporter mutants of Arabidopsis provides evidence that a non-mycorrhizal plant acquires organic nitrogen from agricultural soil

Although organic nitrogen (N) compounds are ubiquitous in soil solutions, their potential role in plant N nutrition has been questioned. We performed a range of experiments on Arabidopsis thaliana genetically modified to enhance or reduce root uptake of amino acids. Plants lacking expression of the Lysine Histidine Transporter 1 (LHT1) displayed significantly lower contents of 13 C and 15 N label and of U-13 C5 ,15 N2 L-glutamine, as determined by liquid chromatography-mass spectrometry when growing in pots and supplied with dually labelled L-glutamine compared to wild type plants and LHT1-overexpressing plants. Slopes of regressions between accumulation of 13 C-labelled carbon and 15 N-labelled N were higher for LHT1-overexpressing plants than wild type plants, while plants lacking expression of LHT1 did not display a significant regression between the two isotopes. Uptake of labelled organic N from soil tallied with that of labelled ammonium for wild type plants and LHT1-overexpressing plants but was significantly lower for plants lacking expression of LHT1. When grown on agricultural soil plants lacking expression of LHT1 had the lowest, and plants overexpressing LHT1 the highest C/N ratios and natural δ15 N abundance suggesting their dependence on different N pools. Our data show that LHT1 expression is crucial for plant uptake of organic N from soil.

Soil diffusive fluxes constitute the bottleneck to tree nitrogen nutrition in a Scots pine forest

Background and aimsIn nutrient poor environments, plant nitrogen (N) acquisition is governed by soil diffusive fluxes and root uptake capacities. However, the relationship between these two processes is not well understood. We explored a way of comparing the processes, enabling identification of the limiting factor for tree N acquisition.MethodsThe study comprised N-fertilized and N-limited Scots pine stands, and measurements of uptake capacities of detached tree roots and of induced soil diffusive fluxes (through in-situ microdialysis) done at the onset and the end of the growing season.ResultsSoil N fluxes were higher at the onset than at the end of the growing season and amino acids comprised a larger fraction of N than inorganic N. N fertilization reduced root uptake capacities of NH4+, glycine and NO3− but not of arginine. For all N compounds except NO3−, diffusive fluxes were significantly lower than root N uptake capacities.ConclusionsOur results suggest that soil N supply in both, N-fertilized and N-limited forest stands, is dominated by amino acids, thus being the major component of plant-available N. Uptake of N appears more constrained by the diffusive fluxes of N compounds rather than root uptake capacity, except for NO3−.