Aims: To investigate the relationship between soil water holding capacity (WHC) and biodegradation of polyester polyurethane (PU) and to quantify and identify the predominant degrading micro-organisms in the biofilms on plastic buried in soil. Methods and Results: High numbers of both fungi and bacteria were recovered from biofilms on soil-buried dumb-bell-shaped pieces of polyester PU after 44 days at 15-100% WHC. The tensile strength of the polyester PU was reduced by up to 60% over 20-80% soil WHC, but no reduction occurred at 15, 90 or 100% soil WHC. A PU agar clearance assay indicated that fungi, but not bacteria were, the major degrading organisms in the biofilms on polyester PU and 10-30% of all the isolated fungi were able to degrade polyester PU in this assay. A 5.8S rDNA sequencing identified 13 strains of fungi representing the three major colony morphology types responsible for PU degradation. Sequence homology matches identified these strains as Nectria gliocladioides (five strains), Penicillium ochrochloron (one strain) and Geomyces pannorum (seven strains). Geomyces pannorum was the predominant organism in the biofilms comprising 22-100% of the viable polyester PU degrading fungi. Conclusions: Polyester PU degradation was optimum under a wide range of soil WHC and the predominant degrading organisms were fungi. Significance and Impact of the Study: By identifying the predominant degrading fungi in soil and studying the optimum WHC conditions for degradation of PU it allows us to better understand how plastics are broken down in the environment such as in landfill sites.