In eukaryotes, nuclear genome sizes vary by more than five orders of magnitude. This variation is not related to organismal complexity, and its origin and biological significance are still disputed. One of the open questions is whether genome size has an adaptive role. We tested the hypothesis that genome size has selective significance, using five grassland communities occurring on a gradient of metal pollution of the soil as a model. We detected a negative correlation between the concentration of contaminating metals in the soil and the number of vascular plant species. Analysis of genome sizes of 70 herbaceous dicot perennial species occurring on the investigated plots revealed a negative correlation between the concentration of contaminating metals in the soil and the proportion of species with large genomes in plant communities. Consistent with the hypothesis, these results show that species with large genomes are at selective disadvantage in extreme environmental conditions.

Combining genotoxicity/mutagenicity tests and physico-chemical methodologies can be useful for determining the potential genotoxic contaminants in soil samples. The aim of our study was to evaluate the genotoxicity of soil by applying an integrated physico-chemical-biological approach. Soil samples were collected at six sampling points in a Slovenian industrial and agricultural region where contamination by heavy metals and sulphur dioxide (SO(2)) are primarily caused by a nearby power plant. The in vitro alkaline version of the comet assay on water soil leachates was performed with Caco-2 and HepG2 cells. A parallel genotoxicity evaluation of the samples was performed by Ames test using Salmonella typhimurium and the Tradescantia micronucleus test. Pedological analyses, heavy metal content determination, and different physico-chemical analyses, were also performed utilizing standard methodology. Water leachates of soil samples were prepared according to standard methods. Since only a battery of biotests with prokaryotic and eukaryotic organisms or cells can accurately estimate the effects of (geno)toxicants in soil samples and water soil leachates, a combination of three bioassays, with cells or organisms belonging to different trophic levels, was used. Genotoxicity of all six water soil leachates was proven by the comet assay on both human cell lines, however no positive results were detected by bacterial assay, Ames test. The Tradescantia micronucleus assay showed increase in micronuclei formation for three samples. According to these results we can assume that the comet assay was the most sensitive assay, followed by the micronucleus test. The Ames test does not appear to be sensitive enough for water soil leachates genotoxicity evaluations where heavy metal contamination is anticipated.

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The response of plant communities to pollution associated with the lead smelter in Zerjav, Slovenia, was investigated on spatial and temporal scales. In 2001, the total concentrations of contaminating metals in the soil measured at the most polluted plot were 59000 mg kg(-1) Pb, 180 mg kg(-1) Cd, and 3300 mg kg(-1) Zn. A negative correlation between the concentration of metals in the soil and plant biodiversity parameters along the gradient of pollution in 2001 was detected. Plant species lists were compiled in 2001 for plots located at different distances from the emission source and compared to that of 1981. In the period from 1981 to 2001, smelter emissions were reduced, and plant species richness increased at all examined plots. Among the successful survivals were some metal hyperaccumulators (Minuartia gerardii, Thlaspi praecox, and Biscutella laevigata). Of special interest were plants that survived the period of highest pollution. We believe that these species can be used in metal-degraded environments for natural revegetation to immobilize heavy metals. The ecosystem in the surroundings of the former smelter is presently recovering. Our results suggest that high metal concentrations in soil are a potential limiting factor for revegetation.

Physicochemical analyses of polluted soils are limited in their ability to determine all hazardous compounds, their bioavailability, and their combined effects on living organisms. Bioassays, on the other hand, can evaluate environmental quality more accurately. This study assesses the genotoxic potential of water extracts from soil polluted with metals (Pb, Cd, and Zn) by the former lead smelter in zerjav, Slovenia using comet assay with Tetrahymena thermophila and human hepatoma cells (HepG2). In addition, the toxicity of soil samples and their extracts was evaluated using Vibrio fischeri and delayed fluorescence of Lemna minor. Chemical analyses of metals using atomic absorption spectrophotometry (AAS) was performed for comparison. Measurements of the total metal concentrations showed that four of five plots near the former lead smelter were highly contaminated with Pb, Cd, and Zn, but the amount of metals in water/soil extracts was low at all the sampling plots. Genotoxicity was demonstrated using T. thermophila for the majority of the extracts, and HepG2 cells for only some of the extracts. Whereas V. fischeri indicated a gradual decrease in soil toxicity with greater distance from the smelter, the toxicity of extracts did not correlate with proximity. Low concentrations of metals in water extracts stimulated L. minor growth. The results indicate that comet assay with T. thermophila and HepG2 cells and the BSPT with V. fischeri are suitable protocols for screening the genotoxic and toxic potential of water/soil extracts by comet assay, whereas chemical analyses of total metal concentrations in soil do not solely suffice for evaluating metal pollution in the environment. Biological assays are thus crucial for risk assessment.