Detection and localization of aluminum and heavy metals in ectomycorrhizal Norway spruce seedlings

Norway spruce seedlings colonized with Hebeloma crustuliniforme were grown in growth pouches. After formation of ectomycorrhizas, the seedlings were exposed to Al or the heavy metals Cd, Cu, Ni, or Zn at various concentrations for 5 weeks to estimate the detection limits of metals with X-ray microanalysis in the cryo-scanning electron microscope. When the lowest metal concentrations (1 mM Al(3+), 0.1 mM Cd(2+), 0.2 mM Cu(2+), 0.5 mM Ni(2+), 2 mM Zn(2+)) were applied, only Al and Zn were detected at low X-ray counts in the ectomycorrhizas. After application of 10-fold higher metal concentrations, distinct metal accumulation patterns were observed. Cd was detected predominantly in the Hartig net, Al and Ni in the Hartig net and in the cell walls of the cortex, and Zn in the Hartig net, the cortical cell walls and the fungal mantle. Cu was not detected at all. By combining X-ray microanalysis with absolute metal concentrations found in the roots, the estimated detection limits of X-ray microanalysis were: Al> or =0.86 mg g(-1), Cd> or =0.26 mg g(-1), Ni> or =1.30 mg g(-1), and Zn> or =0.54 mg g(-1), whereas Cu was not detectable even at root concentrations of 0.47 mg g(-1). Treatments with the highest metal concentrations showed high X-ray counts of metals in cells of the stele but reduced concentrations of the macronutrients K, Mg, and P in roots, indicating a possible disturbance of root and ectomycorrhizal function.