Objectives: Macrofungi substrate specificity and phenology are essential considerations for management of forest ecosystems. In this study, substrate specificity and phenology of macrofungi belonging to the Basidiomycota and Ascomycota groups collected at the University of Dar es Salaam (UDSM) main campus, Tanzania were examined. Methodology and results: Macrofungi were collected throughout the campus during surveys done thrice per year (two rainy seasons, March-May, September- November, and one dry season July- August) from much 2008 to August 2011. Eight traditional morpho-groups based on fruiting bodies comprising more than 67 species distributed in 31 genera in 20 families were collected. The substrate specificity general trend showed that fungi prefer certain specific substrates. Puff fungi are restricted to the soil, fleshy fungi of the Lyophyllaceae on termite mounds soil, and jelly fungi restricted to tree logs although Agarics utilized all the substrates except the wood debris. Tree log substrate supported more macrofungi (28%) followed by soil (26%) and decaying leaf litter (22%). The live tree substrate supported least macrofungi (6%) followed by Wood debris (7%). The general phenology showed that polyporaceae and Ganodermataceae fruit bodies were the most frequently encountered throughout the year. The small sized species from litter-inhabiting genera such as Coprinus, Marasmius and Mycena fruited first with early rains compared to large sized fruit bodies. Conclusion and application of findings: The observation of early fructification relative to fruit body sizes supports the well known finding in the laboratory that the stimulation of fructification is preceded by a marked increase of vegetative growth for mycelia accumulation before being triggered to fruit. According to these study results, this phenomenon seems also important in situ to initiate fruiting of macrofungi. The outcome of this study will contribute to the mycological database for further research and widen the knowledge of biodiversity and substrate relationship which is an efficient parameter in establishing priority for evaluation, utilization and conservation for sustainable forest ecosystem management.Swedish International Development Cooperation Agency (Sida-SAREC)