The aim of the doctoral dissertation was: i) to investigate the tolerance and tolerance mechanisms of three dark septate endophyte (DSE) isolates, Cad#148 and Cad#149 from the genus Cadophora and isolate Leo#146 from the genus Leohumicola to high concentrations of Cd, Zn and Pb in Melin-Norkrans (MNM), and ii) to evaluate their suitability for remediation of contaminated substrate with goat willow (Salix caprea) in combination with different types of undergrowth. Isolates from the genus Cadophora formed 1,8-dihydroxynaphthalene (DHN) melanin in mycelium, with isolate Cad#148 showing higher synthesis. Isolate Leo#146 was not melanized. The sensitivity of isolates to metals was highest for Cd, followed by Zn and Pb, with the melanized isolates showing higher tolerance due to lower accumulation of metals in the mycelium and lower oxidative stress. Organic acid synthesis was increased in the presence of Cd and Zn stress, although the type of organic acids induced was isolate-specific. Melanin synthesis in melanized isolates decreased with increasing metal stress. However, by using a melanin synthesis inhibitor in the presence of Cd stress, we demonstrated that melanin plays an important role in the tolerance of the most melanized DSE isolate Cad#148 to Cd. Furthermore, in a pot experiment, we investigated the influence of inoculation with selected DSE isolates and different types of undergrowth (natural, sown) on the physiology and accumulation of metals in willow rooted in non-sterile contaminated substrate. DSE isolates Leo#146 and Cad #148 were most successful in infecting willow roots and had positive affect on growth and synthesis of total chlorophyll and carotenoid pigments in willow leaves. Inoculation of willows with the DSE isolates Leo#146 in combination with undergrowth blue moor grass reduced the transfer of Cd and Zn from roots to leaves and consequently accumulation of Cd and Zn in leaves. In addition, inoculation of willow cuttings with Leo#146 and Cad#148 in combination with undergrowth Rožnik reduced the translocation of Pb from roots to leaves. The DSE isolate Leo#146 in combination with the undergrowth of blue moor grass was selected as the most suitable isolate for phytoremediation of contaminated substrate with willow.
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