Significance of Silver Birch and Bushgrass for Establishment of Microbial Heterotrophic Community in a Metal-Mine Spoil Heap

Abstract

Differences in the culturable fractions of total and metal-tolerant bacteria inhabiting bulk soil of a metal-mine spoil heap and the rhizosphere of silver birch (Betula pendula) or bushgrass (Calamagrostis epigejos), completed with changes in total microbial community structure in the soil, were assessed by MIDI-FAME (fatty acid methyl ester) profiling of whole-cell fatty acids. In addition, the abundance of metal-tolerant populations among the culturable bacterial communities and their identity and the metal-tolerance patterns were determined. The high proportions of Cu- and Zn-tolerant bacteria that ranged from 60.6% to 94.8% were ascertained in the heap sites. Within 31 bacterial isolates obtained, 24 strains were Gram-positive and Arthrobacter, Bacillus, Rathayibacter, Brochothrix, and Staphylococcus represented those identified. Minimum inhibitory concentration (MIC) data indicated that several strains developed multi-metal tolerance, and the highest tolerance to Cu (10 mM) and Zn (12 mM) was found for Pseudomonas putida TP3 and three isolated strains (BS3, TP12, and SL16), respectively. The analysis of FAME profiles obtained from the culturable bacterial communities showed that Gram-positive bacteria predominated in bulk soil of all heap sites. In contrast, the rhizosphere communities showed a lower proportion of the Gram-positive group, especially for silver birch. For the total microbial community, mostly Gram-negative bacteria (e.g., Pseudomonas) inhabited the heap sites. The results suggest that the quantitative and qualitative development of heterotrophic microbiota in the soil of the metal-mine spoil heap seems to be site-dependent (i.e., rhizosphere vs. bulk soil), according to differences in the site characteristics (e.g., enrichment of nutrients and total metal concentrations) and impact of plant species.