In this study we explored leaf litter fungal diversity in a diverse Panama lowland tropical forest in which a replicated factorial N, P, K, and micronutrient fertilization experiment of 40 x 40 m plots had been on-going for nine years. We extracted DNA from leaf litter samples and used fungal-specific amplification and a 454 pyrosequencing approach to sequence two loci, the nuclear ribosomal internal transcribed spacer (ITS) region and the nuclear ribosomal large subunit (LSU) D1 region.