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In forested headwater streams, decomposition of allochthonous organic matter is a fundamental process driven by aquatic microbes and invertebrate shredders. We examined how season and eutrophication affect leaf decomposition and the associated decomposer communities by immersing leaves of a late deciduous species (Quercus robur) in five streams in Portugal along a gradient of eutrophication in autumn and spring. We found hump-shaped relationships between leaf decomposition and total nitrogen and phosphorus in stream water in both seasons. Leaf decomposition and shredder biomass were higher during spring in streams with moderate levels of eutrophication. Fungal sporulation and biomass were stimulated at moderate levels of eutrophication and inhibited at low or high levels of eutrophication. Fungal assemblage composition shifted between seasons and along the gradient of eutrophication. Tricladium chaetocladium increased its contribution to total conidial production in spring, while Dimorphospora foliicola was dominant in the most eutrophic streams where Articulospora tetracladia was almost absent. Invertebrate shredders were the primary decomposers of leaves in streams with moderate levels of eutrophication, particularly in the warmest season. Although the presence of late deciduous plant species, such as oak, in the riparian corridors may help to mitigate food depletion to freshwater decomposers in spring, our results suggest that moderate eutrophication can accelerate decomposition further reducing litter standing stocks in the warmer seasons.

a b s t r a c tWe assessed aquatic hyphomycete diversity in autumn and spring on oak leaves decomposing in five streams along a gradient of eutrophication in the Northwest of Portugal. Diversity was assessed through microscopy-based (identification by spore morphology) and DNA-based techniques (Denaturing Gradient Gel Electrophoresis and 454 pyrosequencing). Pyrosequencing revealed five times greater diversity than DGGE. About 21% of all aquatic hyphomycete species were exclusively detected by pyrosequencing and 26% exclusively by spore identification. In some streams, more than half of the recorded species would have remained undetected if we had relied only on spore identification. Nevertheless, in spring aquatic hyphomycete diversity was higher based on spore identification, probably because many species occurring in this season are not yet connected to ITS barcodes in genetic databases. Pyrosequencing was a powerful tool for revealing aquatic hyphomycete diversity on decomposing plant litter in streams and we strongly encourage researchers to continue the effort in barcoding fungal species.

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