Ultrastructural element localization by EDXS in Empetrum nigrum.

Empetrum nigrum L. is one of the few species growing on highly polluted areas in the northern boreal forests and it accumulates considerable amounts of heavy metals especially in its older stems. Previous-year stems of Empetrum nigrum were collected from two different sites located at distances of 0.5km (highly contaminated) and 8km (low contaminated) from a Cu--Ni smelter at Harjavalta, SW Finland. The element (Al, As, Cu, Fe, Mn, Zn, Ca, K, P, S, Mg, Na) localization was performed by energy-dispersive X-ray spectroscopy (EDXS) after cryofixation, freeze-drying and pressure infiltration of the material. The results showed higher levels of Cu, As and Fe in cell compartments of E. nigrum close to the smelter than at further distance. The Al and Zn levels, in contrast, showed no clear differences between the sites. Cu was distributed homogeneously in the tissue and occurred in vacuoles, cytoplasm, cell walls as well as in lumens of the vascular tissue. The higher amounts of As were localized in the outer regions of the stem cross-section and the amounts were higher in the primary cell walls of living (ray cells, phloem) than dead cells (xylem, sclereids). Ray cells, phloem and sclereids had elevated Fe amounts compared to the other tissues in the contaminated stem samples but owing to the high variation between the replicates, no significant differences were found. Based on the rather homogeneous localization of Cu, As and Fe in the living tissue and increased levels of Cu, As and Fe in vacuoles, cell walls and cytoplasm near the smelter, it seems that more than one specific mechanism contribute to the heavy metal tolerance of E. nigrum. Macronutrients did not show clear differences between the two distances or connection to heavy metal localization. Neither the role of complexing agents in heavy metal tolerance in the cytoplasm or vacuoles could be shown by this study. Because of the more frequent localization of electron dense phenolic material in the polluted samples, it might also have a function in the heavy metal tolerance of E. nigrum.