Two ectomycorrhizal truffles, Tuber melanosporum and T. aestivum, endophytically colonise roots of non-ectomycorrhizal plants in natural environments.

Serendipitous findings and studies on Tuber species suggest that some ectomycorrhizal fungi, beyond their complex interaction with ectomycorrhizal hosts, also colonise roots of nonectomycorrhizal plants in a loose way called endophytism. Here, we investigate endophytism of T. melanosporum and T. aestivum. We visualised endophytic T. melanosporum hyphae by fluorescent in situ hybridisation on nonectomycorrhizal plants. For the two Tuber species, microsatellite genotyping investigated the endophytic presence of the individuals whose mating produced nearby ascocarps. We quantified the expression of four T. aestivum genes in roots of endophyted, non-ectomycorrhizal plants. Tuber melanosporum hyphae colonised the apoplast of healthy roots, confirming endophytism. Endophytic Tuber melanosporum and T. aestivum contributed to nearby ascocarps, but only as maternal parents (forming the flesh). Paternal individuals (giving only genes found in meiotic spores of ascocarps) were not detected. Gene expression of T. aestivum in non-ectomycorrhizal plants confirmed a living status. Tuber species, and likely other ectomycorrhizal fungi found in nonectomycorrhizal plant roots in this study, can be root endophytes. This is relevant for the ecology (brûlé formation) and commercial production of truffles. Evolutionarily speaking, endophytism may be an ancestral trait in some ectomycorrhizal fungi that evolved from root endophytes.

Soil spore bank in Tuber melanosporum: up to 42% of fruitbodies remain unremoved in managed truffle grounds.

Fungi fruiting hypogeously are believed to form spore banks in soil especially because some fruitbodies are not removed by animals. However, little is known on the proportion of fruitbodies that are not removed by animals. We took advantage of the brûlé phenomenon, which allows delineation of the mycelium distribution, to assess the proportion of unremoved black truffle (Tuber melanosporum) fruitbodies in the context of plantations where fruitbodies are actively sought and harvested by truffle growers. We inspected portions of the brûlés after the harvest season to find unremoved fruitbodies. On average, from six truffle grounds in which a total of 38 brûlés were investigated, unremoved fruitbodies represented 33% of the whole fruitbody production (42% when averaging all the brûlés). We discuss this value and its high variability among truffle grounds. Beyond the local and variable accidental reasons that may lead to this high proportion, we speculate that the formation of some undetectable fruitbodies may be under selection pressure, given the reproductive biology of T. melanosporum.

Effect of slug mycophagy on Tuber aestivum spores.

Truffles in the genus Tuber produce subterranean fruiting bodies that are not able to actively discharge their spores in the environment. For this reason, truffles depend on mycophagous animals for reproduction. Fungus consumption (mycophagy) is a behaviour typical of both vertebrates and invertebrates. Mammals, especially rodents, are the most studied group of mycophagists and have been found to consume a great variety of fungi. Among invertebrates, mycophagy is documented in arthropods, but rarely in molluscs. In our study we assessed the effect on the morphology and mycorrhizal colonization of Tuber aestivum spores after passage through the gut of slugs (Deroceras invadens) and, for comparison, of a house mouse (Mus musculus). Light, scanning electron and atomic force microscopy revealed that the digestion, especially by slugs, freed spores from the asci and modified their morphology. These are believed to be the reasons why we observed an improvement in oak mycorrhization with the slug and rodent ingested spores in comparison to a fresh spore inoculation. We also demonstrated by molecular barcoding that slugs' guts sampled on a Tuber melanosporum truffle ground contain spores from this species and Tuber brumale, further suggesting that some invertebrates are efficient Tuber spore dispersers.

Colonization and extinction dynamics among the plant species at tree bases in Paris (France).

In cities, trees planted along streets could play an important ecological role for spontaneous plants growing at their bases. For example, these trees could represent corridors by potentially connecting large green spaces (e.g., parks, gardens), which allow species to move within the urban matrix. We considered sets of urban trees in 15 streets in Paris, France, as metapopulations for 15 plant species. Our objective was to determine the factors influencing the dynamics of colonization and extinction of populations based on the distance of the streets to green spaces and biological traits of each species.Plant species in 1,324 tree bases of the Bercy District of Paris were surveyed annually from 2009 to 2015. For each species and each street, we used SPOMSIM software to identify the best-fit metapopulation model between four models with different colonization and extinction functions: propagule rain model (PRM) and Levins' model with or without rescue effect.Results demonstrated that species more often conformed to the PRM in streets near green spaces, which suggested that green spaces could act as sources for the populations in those streets. Species with seeds with long-term persistence more often conformed to the PRM, indicating that a soil seed bank helps species invade entire streets. Finally, a higher percentage of species with a short height conformed to models with a rescue effect, which indicated that those small species resisted the effects of weeding by the city technical services better than taller species.Synthesis and applications. This study showed how biological traits of species and geography of the district determine the dynamics of plants in the streets, and these results may provide important information for biodiversity management in cities.

Black Truffle, a Hermaphrodite with Forced Unisexual Behaviour.

The life cycle of the black truffle (Tuber melanosporum) includes a mating before sporulation: although the species is hermaphroditic, mating turns out to involve parents with very different features, that mostly behave as male or female only, suggesting that this species undergoes forced dioecism.