Heavy Metal Tolerance and Accumulation Potential of a Rare Coastal Species, Anthyllis vulneraria subsp. maritima

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Abstract
The aim of the present study was to explore heavy metal tolerance and accumulation potential in Anthyllis vulneraria subsp. maritima plants from coastal sand dunes in controlled conditions. Plants were established from seeds collected in coastal sand dunes and cultivated in substrates in greenhouse conditions. A gradual treatment with CdCl2, PbOAc, CuSO4, MnSO4, and ZnSO4 was performed until three final concentrations for each metal were reached. The number of leaves, their biomass, and biomass of roots were negatively affected by increasing concentrations of lead (Pb) and manganese (Mn) in substrate, but no negative effect was evident for cadmium (Cd), copper (Cu), and zinc (Zn). Visible effects of metal toxicity were evident for Pb-treated plants (appearance of thinner leaves, yellowing of older leaves), as well as for Mn-treated plants (reduced leaf size, curled leaves, red leaf venation). There was a significant decrease in water content in old leaves at high Pb and increasing Mn concentration, indicating accelerated leaf senescence. Increase in polyphenol oxidase activity in leaves was evident in all the plants treated with heavy metals. In contrast, an increase in peroxidase activity was evident only for plants treated with 50 and 100 mg L−1 Cd, 500 mg L−1 Pb, 200–1000 mg L−1 Mn, and 500 mg L−1 Zn. Metal accumulation potential for Cd and Cu was the highest in the roots, but for Pb, Mn, and Zn, more metal accumulated in old leaves. It can be concluded that A. vulneraria subsp. maritima plants are tolerant to high Cd, Cu, and Zn, but moderately susceptible to Pb and Mn. However, oxidative enzyme activity cannot be unequivocally used as a specific indicator of metal tolerance. In respect to phytoremediation potential, the plants have very good accumulation capacity for Pb, Mn, and Zn.

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