Shifts in community composition provide a mechanism for maintenance of activity of soil yeasts in the presence of elevated copper levels

Abstract

Soil dilution plates were prepared from different soil samples using a solid synthetic selective medium containing (i). glucose as carbon source, (ii). thymine as nitrogen source, (iii). vitamins, (iv). minerals, and (v). chloramphenicol as antibacterial agent. Using the Diazonium Blue B colour reaction, it was found that both ascomycetous and basidiomycetous yeasts were able to grow on this medium. Subsequently, the medium was used to enumerate yeasts in soil microcosms prepared from four different soil samples, which were experimentally treated with the fungicide copper oxychloride, resulting in copper (Cu) concentrations of up to 1000 ppm. The selective medium supplemented with 32 ppm of Cu was used to enumerate Cu-resistant yeasts in the microcosms. The results showed that the addition of Cu at concentrations >or=approximately 1000 ppm did not have a significant effect on total number of yeasts in the soil. Furthermore, it was found that Cu-resistant yeasts were present in all the soil samples, regardless of the amount of Cu that the soil was challenged with. At the end of the incubation period, yeasts in the microcosms with zero and approximately 1000 ppm of additional Cu were enumerated, isolated, and identified with sequence analyses of the D1/D2 600-650 bp region of the large subunit of ribosomal DNA. Hymenomycetous species dominated in the control soil, while higher numbers of the urediniomycetous species were found in the soil that received Cu. These observations suggest that urediniomycetous yeasts may play an important role in re-establishing overall microbial activity in soils, following perturbations, such as the addition of Cu-based fungicides.