Integrated approach to assessing the effects of soils polluted with heavy metals on a plant population

Abstract
This study addresses the effects of soil polluted with more than one heavy metal in a grass species. A 16-week bioassay with Avena sativa L. was conducted in microcosms using soils from two abandoned mines in central Spain that contained levels above World Health Organization (WHO) reference limits for soils of more than three heavy metals. Pollution effects were examined at cell, tissue, organ, plant and population levels. For this purpose, dry weight, maximum height and number of leaves were determined; leaf tissues were observed by low temperature scanning electron microscopy; the metal contents of roots and shoots were determined by plasma emission spectroscopy and their distribution in different tissues was analyzed by X-ray microanalysis using an environmental scanning electron microscope. The results explain the accumulation and translocation of soil metals by this plant species; their effects in cells, tissues and growth of plants; and allow inference on population effects. The discussion of the methodological approach leads us to propose a valid protocol to assess the effects of a set of heavy metals present in the topsoil of polluted sites on a plant population. We recommend its use for an ecotoxicological diagnosis and risk analysis of similarly polluted sites.