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The decomposition of plant litter is a major process of equivalent status to primary production in ecosystem functioning. The spatiotemporal changes in the composition and dynamics of litter fungal community along a climate gradient ranging from arid desert to humid-Mediterranean regions in Israel was examined using wheat straw litter bags placed at four selected sites along the climate gradient, arid, semi-arid, Mediterranean, and humid-Mediterranean sites. Litter samples were collected over a two-year decomposition period to evaluate litter weight loss, moisture, C:N ratio, fungal composition, and isolate density. The litter decomposition rate was found to be the highest during the first year of the study at the Mediterranean and arid sites. Although the Shannon-Wiener index values of the fungal communities in the litter samples were the highest at the humid-Mediterranean site, the number of fungal species was not significantly different between the four study sites. Different fungal groups were found to be related to different study sites: Basidiomycota, Mucoromycotina, and teleomorphic Ascomycota were associated with the humid-Mediterranean site, while Coelomycetes were mostly affected by the arid site. Our results indicate that climate factors play an important role in determining the structure of saprotrophic fungal communities in the decomposing litter and in mediating plant litter decomposition processes.