Summary• Root colonization by arbuscular mycorrhizal fungi (AMF) was investigated in industrially polluted grassland characterized by exceptionally high phosphorus levels (up to 120 g kg − 1 soil).• Along a pollution-induced nitrogen gradient, soil and tissue element concentrations of Artemisia vulgaris plants and their mycorrhizal status were determined. Additionally, we compared mycorrhization rates and above-ground biomass of A. vulgaris at N-fertilized and control plots in the N-poor area.• Despite high soil and tissue P concentrations, plants from N-deficient plots, which were characterized by low tissue N concentrations and N : P ratios, were strongly colonized by AMF, whereas at a plot with comparable P levels, but higher soil and plant N concentrations and N : P ratios, mycorrhization rates were significantly lower. Correlation analyses revealed a negative relationship between percentage root colonization of A. vulgaris by AMF and both tissue N concentration and N : P ratio. Accordingly, in the fertilization experiment, control plants had higher mycorrhization rates than N-fertilized plants, whereas the species attained higher biomass at Nfertilized plots.• The results suggest that N deficiency stimulates root colonization by AMF in this extraordinarily P-rich field site.

A method has been developed for quantification of 20 amino acids as well as 13 (15)N-labeled amino acids in barley plants. The amino acids were extracted from plant tissues using aqueous HCl-ethanol and directly analyzed without further purification. Analysis of the underivatized amino acids was performed by liquid chromatography (LC)-electrospray ionization (ESI) tandem mass spectrometry (MS-MS) in the positive ESI mode. Separation was achieved on a strong cation exchange column (Luna 5micro SCX 100A) with 30 mM ammonium acetate in water (solvent A) and 5% acetic acid in water (solvent B). Quantification was accomplished using d (2)-Phe as an internal standard. Calibration curves were linear over the range 0.5-50 microM, and limits of detection were estimated to be 0.1-3.0 microM. The mass-spectrometric technique was employed to study the regulation of amino acid levels in barley plants grown at 15 degrees C uniform root temperature (RT) and 20-10 degrees C vertical RT gradient (RTG). The LC-MS-MS results demonstrated enhanced concentration of free amino acids in shoots at 20-10 degrees C RTG, while total free amino acid concentration in roots was similarly low for both RT treatments. (15)NO(3) (-) labeling experiments showed lower (15)N/(14)N ratios for Glu, Ser, Ala and Val in plants grown at 20-10 degrees C RTG compared with those grown at 15 degrees C RT.

We have detailed knowledge from controlled environment studies on the influence of root temperature on plant performance, growth and morphology. However, in all studies root temperature was kept spatially uniform, which motivated us to test whether a vertical gradient in soil temperature affected development and biomass production. Roots of barley seedlings were exposed to three uniform temperature treatments (10, 15 or 20°C) or to a vertical gradient (20-10°C from top to bottom). Substantial differences in plant performance, biomass production and root architecture occurred in the 30-day-old plants. Shoot and root biomass of plants exposed to vertical temperature gradient increased by 144 respectively, 297%, compared with plants grown at uniform root temperature of 20°C. Additionally the root system was concentrated in the upper 10 cm of the soil substrate (98% of total root biomass) in contrast to plants grown at uniform soil temperature of 20°C (86% of total root biomass). N and C concentrations in plant roots grown in the gradient were significantly lower than under uniform growth conditions. These results are important for the transferability of 'normal' greenhouse experiments where generally soil temperature is not controlled or monitored and open a new path to better understand and experimentally assess root-shoot interactions.

The ability of a quadrupole-based ICP-MS with an octopole collision cell to obtain precise and accurate measurements of isotope ratios of magnesium, calcium and potassium was evaluated. Hydrogen and helium were used as collision/reaction gases for ICP-MS isotope ratio measurements of calcium and potassium in order to avoid isobaric interference with the analyte ions from (mainly) argon ions 40Ar+ and argon hydride ions 40Ar1H+. Mass discrimination factors determined for the isotope ratios 25Mg/24Mg, 40Ca/44Ca and 39K/41K under optimized experimental conditions varied between 0.044 and 0.075. The measurement precisions for 25Mg/24Mg, 40Ca/44Ca and 39K/41K were found to be 0.09%, 0.43% and 1.4%, respectively. This analytical method that uses ICP-QMS with a collision cell to obtain isotope ratio measurements of magnesium, calcium and potassium was used in routine mode to characterize biological samples (nutrient solution and small amounts of digested plant samples). The mass spectrometric technique was employed to study the dynamics of nutrient uptake and translocation in barley plants at different root temperatures (10 degrees C and 20 degrees C) using enriched stable isotopes (25Mg, 44Ca and 41K) as tracers. For instance, the mass spectrometric results of tracer experiments demonstrated enhanced 25Mg and 44Ca uptake and translocation into shoots at a root temperature of 20 degrees C 24 h after isotope spiking. In contrast, results obtained from 41K tracer experiments showed the highest 41K contents in plants spiked at a root temperature of 10 degrees C.