We used different tests to assess the effect of high soil concentrations of heavy metals on pollen viability in plants from metallicolous (MET) and nonmetallicolous (NONMET) populations. The frequency of viable pollen depended on the test applied: MET plants showed no significant reduction of pollen viability by acetocarmine, Alexander, MTT and X-Gal dye testing, but a drastic reduction of pollen viability in MET flowers (MET 56% vs 72% NONMET) by the FDA test. There was no correlation between pollen viability estimated in histochemical tests and pollen germination in vitro or in vivo. We discuss the terminology used to describe pollen viability as determined by histochemical tests.K Ke ey y w wo or rd ds s: : Viola tricolor, pollen viability tests, pollen germination, metalliferous sites, heavy metals, acetocarmine test, Alexander test, X-Gal test, FDA test, MTT test.
Intraspecific changes in genome size and chromosome number lead to divergence and species evolution. Heavy metals disturb the cell cycle and cause mutations. Areas contaminated by heavy metals (metalliferous sites) are places where microevolutionary processes accelerate; very often only a few generations are enough for a new genotype to arise. This study, which continues our long-term research on Viola tricolor (Violaceae), a species occurring on both metalliferous (Zn, Pb, Cd, Cu) and non-metalliferous soils in Western and Central Europe, is aimed at determining the influence of environments polluted with heavy metals on genome size and karyological variability. The genome size of V. tricolor ranged from 3.801 to 4.203 pg, but the differences between metallicolous and non-metallicolous populations were not statistically significant. Altered chromosome numbers were significantly more frequent in material from the polluted sites than from the non-polluted sites (43% versus 28%). Besides the standard chromosome number (2n = 26), aneuploid cells with lower (2n = 18-25) or higher (2n = 27, 28) chromosome numbers were found in plants from both types of site, but polyploid (2n = 42) cells were observed only in plants from the metalliferous locality. The lack of correlation between chromosome variability in root meristematic cells and genome size estimated from peduncle cells can be attributed to elimination of somatic mutations in generative meristem, producing chromosome-stable non-meristematic tissues in the peduncle.
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