Phylogenetic relationships of the genera Russula and Lactarius were investigated using sequence data from the nuclear-encoded large subunit ribosomal DNA (LSU rDNA). Ninety-five sequences belonging to the genera Russula and Lactarius, including 31 sequences from the databases, were used in this study. Analysis of the LSU rDNA region indicated that Russulaceae was divided into six groups (group A-F) in the neighbor-joining (NJ) tree. Lactarius consisted of one large clade (group A). Therefore, this genus was found to be monophyletic. However, the monophyly of genus Russula remained unclear. The genus Russula consisted of five groups in the NJ tree. Group B includes sects. Plorantes and Archaeinae (Heim), and group C includes sects. Delicoarchaeae and Russula in the NJ tree. Neither of the two groups formed a single clade in the most parsimonius (MP) tree. Group D includes many taxa having colored spore prints and amyloid in suprahilar plage of spores in sect. Russula and sect. Rigidae. Group E consists of only sect. Compactae and is further divided into three subclades, represented by R. densifolia, R. nigricans, and R. subnigricans, respectively. Group F contains sects. Rigidae, Ingratae, and Pelliculariae. Sect. Compactae and sect. Plorantes should not be as closely related as previously supposed. Russula earlei may be placed in sect. Archaeinae Heim. Russula flavida (subsect. Amoeninae) is placed in sect. Russula with R. aurea with a high bootstrap value (99%). The nuclear LSU rDNA region is a useful tool in recognization of species of Russulaceae and may provide information concerning phylogenetic relationships between the genera Russula and Lactarius.

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Summary Investigation of seasonal variation in fungal communities is essential for understanding biodiversity and ecosystem functions. However, the conventional sampling method, with substrate removal and high spatial heterogeneity of community composition, makes surveying the seasonality of fungal communities challenging. Recently, water environmental DNA (eDNA) analysis has been explored for its utility in biodiversity surveys. In this study, we assessed whether the seasonality of fungal communities can be detected by monitoring eDNA in a forest stream. We conducted monthly water sampling in a forest stream over 2 years and used DNA metabarcoding to identify fungal eDNA. The stream water contained DNA from functionally diverse aquatic and terrestrial fungi, such as plant decomposers, parasites and mutualists. The variation in the fungal assemblage showed a regular annual periodicity, meaning that the assemblages in a given season were similar, irrespective of the year or sampling. Furthermore, the strength of the annual periodicity varied among functional groups. Our results suggest that forest streams may act as a ‘trap’ for terrestrial fungal DNA derived from different habitats, allowing the analysis of fungal DNA in stream water to provide information about the temporal variation in fungal communities in both the aquatic and the surrounding terrestrial ecosystems.