Bioethanol generated from household, agricultural or forestry waste is currently being pursued as the second generation biofuel with wheat straw being highlighted as a promising lignocellulosic residue feedstock in the UK. Natural fermentative ecosystems such as agricultural manure and forest floors containing decomposing wood and leaf litter are potential sources of yeast strains with suitable phenotypic characteristics for improved production of bioethanol. Identifying yeast with innate fermentation capabilities could potentially contain a pool of genes that confer resistance to the chemical constituents of lignocellulosic biomass. Wild yeasts were analysed using a phenotypic microarray for utilization of xylose and tolerance to pre-treatment inhibitors. Assays identified yeast which had stress tolerant phenotypes and yeast with xylose utilization capabilities. Performance fermentations confirmed desirable phenotypes such as conversion of xylose into ethanol and inhibitor tolerance, making these yeasts of interest for further industrial exploration and exploitation.