The article presents the effect of a combined amendment, i.e., biochar+compost (BC), on the process of Cd, Cu, Ni, Pb and Zn immobilization in soil cultivated with L. perenne under freezing and thawing conditions (FTC). In particular, the speciation analysis of the examined elements in phytostabilized soils based on their response using the sequential extraction, and the variability of the soil microbiome using 16S rRNA gene amplicon sequencing were systematically assessed. Metal stability in soils was evaluated by the reduced distribution index (Ir). Plants were grown in pots for 52 days under greenhouse conditions. After termination, phytostabilization was continued in a temperature chamber for 64 days to provide FTC. As a result, it was noted that biomass yield of L. perenne was promoted by BC (39 % higher than in the control pots) and reduced by FTC (45 % lower than in the BC-enriched soil not exposed to FTC). An efficacious level of phytostabilization, i.e., higher content of heavy metals in plant roots, was found in the BC-enriched soil, regardless of the changes in soil temperature conditions. BC improved soil pH before applying FTC more than after applying FTC. BC had the greatest impact on increasing Cu stability by redistributing it from the F1 and F2 fractions to the F3 and F4 fractions. For most metals, phytostabilization under FTC resulted in an increase in the proportion of the F1 fraction and a decrease in its stability. Only for Pb and Zn, FTC had greater impact on their stability than BC addition. In all soil samples, the core genera with about 2-3 % abundances were Sphingomonas sp. and Mycobacterium sp. FTC favored the growth of Bacteroidetes and Proteobacteria in soil. Microbial taxa that coped well with FTC but only in the absence of BC were Rhodococcus, Alkanindiges sp., Flavobacterium sp., Williamsia sp. Thermomonas sp.
Keywords: Combined amendments; Contaminated soil; Immobilization; Low temperatures; Microbiome.
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