Context-dependent benefits of forest soil addition on Aleppo pine seedling performance under drought and grass competition.

Seedling establishment under natural conditions is limited by numerous interacting factors. Here, we tested the combined effects of drought, herbaceous competition, and ectomycorrhizal inoculation on the performance of Aleppo pine seedlings grown in a net-house. The roots of all pine seedlings were strongly dominated by Geopora, a fungal genus known to colonize seedlings in dry habitats. Ectomycorrhizal fungi (EMF) inoculum significantly increased seedling height, biomass, and the number of side branches. However, under either competition or drought, the positive effect of EMF on seedling biomass and height was greatly reduced, while the effect on shoot branching was maintained. Further, under a combination of drought and competition, EMF had no influence on either plant growth or shape. The discrepancy in pine performance across treatments highlights the complexity of benefits provided to seedlings by EMF under ecologically relevant settings.

Belowground carbon transfer across mycorrhizal networks among trees: Facts, not fantasy.

The mycorrhizal symbiosis between fungi and plants is among the oldest, ubiquitous and most important interactions in terrestrial life on Earth. Carbon (C) transfer across a common mycorrhizal network (CMN) was demonstrated over half a century ago in the lab ( Reid & Woods, 1969), and later in the field ( Simard et al., 1997a). Recent years have seen ample progress in this research direction, including evidence for ecological significance of carbon transfer ( Klein et al., 2016). Furthermore, specific cases where the architecture of mycorrhizal networks have been mapped ( Beiler et al., 2015) and CMN-C transfer from mature trees to seedlings has been demonstrated ( Orrego, 2018) have suggested that trees in forests are more connected than once thought ( Simard, 2021). In a recent Perspective, Karst et al. (2023) offered a valuable critical review warning of over-interpretation and positive citation bias in CMN research. It concluded that while there is evidence for C movement among plants, the importance of CMNs remains unclear, as noted by others too ( Henriksson et al., 2023). Here we argue that while some of these claims are justified, factual evidence about belowground C transfer across CMNs is solid and accumulating.

A biome-dependent distribution gradient of tree species range edges is strongly dictated by climate spatial heterogeneity.

Understanding the causes of the arrest of species distributions has been a fundamental question in ecology and evolution. These questions are of particular interest for trees owing to their long lifespan and sessile nature. A surge in data availability evokes a macro-ecological analysis to determine the underlying forces limiting distributions. Here we analyse the spatial distribution of >3,600 major tree species to determine geographical areas of range-edge hotspots and find drivers for their arrest. We confirmed biome edges to be strong delineators of distributions. Importantly, we identified a stronger contribution of temperate than tropical biomes to range edges, adding strength to the notion that tropical areas are centres of radiation. We subsequently identified a strong association of range-edge hotspots with steep spatial climatic gradients. We linked spatial and temporal homogeneity and high potential evapotranspiration in the tropics as the strongest predictors of this phenomenon. We propose that the poleward migration of species in light of climate change might be hindered because of steep climatic gradients.

Asymmetric belowground carbon transfer in a diverse tree community.

Mycorrhizal fungi can colonize multiple trees of a single or multiple taxa, facilitating bidirectional exchange of carbon between trees. Mycorrhiza-induced carbon transfer was shown in the forest, but it is unknown whether carbon is shared symmetrically among tree species, and if not, which tree species are better donors and which are better recipients. Here, we test this question by investigating carbon transfer dynamics among five Mediterranean tree species in a microcosm system, including both ectomycorrhizal (EM) and arbuscular (AM) plants. Trees were planted together in "community boxes" using natural soil from a mixed forest plot that serves as a habitat for all five tree species and their native mycorrhizal fungi. In each box, only the trees of a single species were pulse-labelled with 13 CO2 . We found that carbon transfer was asymmetric, with oak being a better donor, and pistacia and cypress better recipients. Shared mycorrhizal species may have facilitated carbon transfer, but their diversity did not affect the amount, nor timing, of the transfer. Overall, our findings in a microcosm system expose rich, but hidden, belowground interactions in a diverse population of trees and mycorrhizal fungi. The asymmetric carbon exchange among cohabiting tree species could potentially contribute to forest resilience in an uncertain future.

Ectomycorrhizal fungi mediate belowground carbon transfer between pines and oaks.

Inter-kingdom belowground carbon (C) transfer is a significant, yet hidden, biological phenomenon, due to the complexity and highly dynamic nature of soil ecology. Among key biotic agents influencing C allocation belowground are ectomycorrhizal fungi (EMF). EMF symbiosis can extend beyond the single tree-fungus partnership to form common mycorrhizal networks (CMNs). Despite the high prevalence of CMNs in forests, little is known about the identity of the EMF transferring the C and how these in turn affect the dynamics of C transfer. Here, Pinus halepensis and Quercus calliprinos saplings growing in forest soil were labeled using a 13CO2 labeling system. Repeated samplings were applied during 36 days to trace how 13C was distributed along the tree-fungus-tree pathway. To identify the fungal species active in the transfer, mycorrhizal fine root tips were used for DNA-stable isotope probing (SIP) with 13CO2 followed by sequencing of labeled DNA. Assimilated 13CO2 reached tree roots within four days and was then transferred to various EMF species. C was transferred across all four tree species combinations. While Tomentella ellisii was the primary fungal mediator between pines and oaks, Terfezia pini, Pustularia spp., and Tuber oligospermum controlled C transfer among pines. We demonstrate at a high temporal, quantitative, and taxonomic resolution, that C from EMF host trees moved into EMF and that C was transferred further to neighboring trees of similar and distinct phylogenies.

A non-linear effect of the spatial structure of the soil ectomycorrhizal spore bank on the performance of pine seedlings.

The spatial structure of the environment is known to affect ecological processes. Unlike the spatial structure of negative interactions, such as competition and predation, the role of spatial structure in positive interaction has received less attention. We tested how the spatial structure of spores of ectomycorrhizal fungi (EMF) in the soil affects the growth of Aleppo pine (Pinus halepensis) seedlings. Spores were spatially distributed at four different levels of patchiness (1 patch, 4 patches, 8 patches and complete mixing) in 4 L pots (all pots received the same total amount of spores). Based on previous findings, we hypothesized that plant performance would gradually increase from the single patch treatment to the complete mixing. However, we found a non-linear response to patchiness. Specifically, plants were largest in the single patch and complete mixing while those in the 4 and 8 patch treatments were the smallest. This non-monotonic response, which might be the result of spatially determined colonization timing or community composition, suggests that the spatial structure of EMF spores has a complex effect on seedling growth.

High resilience of the mycorrhizal community to prescribed seasonal burnings in eastern Mediterranean woodlands.

Fire effects on ecosystems range from destruction of aboveground vegetation to direct and indirect effects on belowground microorganisms. Although variation in such effects is expected to be related to fire severity, another potentially important and poorly understood factor is the effect of fire seasonality on soil microorganisms. We carried out a large-scale field experiment examining the effects of spring (early-dry season) versus autumn (late-dry- season) burns on the community composition of soil fungi in a typical Mediterranean woodland. Although the intensity and severity of our prescribed burns were largely consistent between the two burning seasons, we detected differential fire season effects on the composition of the soil fungal community, driven by changes in the saprotrophic fungal guild. The community composition of ectomycorrhizal fungi, assayed both in pine seedling bioassays and from soil sequencing, appeared to be resilient to the variation inflicted by seasonal fires. Since changes in the soil saprotrophic fungal community can directly influence carbon emission and decomposition rates, we suggest that regardless of their intensity and severity, seasonal fires may cause changes in ecosystem functioning.

Small-scale spatial variability in the distribution of ectomycorrhizal fungi affects plant performance and fungal diversity.

The effects of spatial heterogeneity in negative biological interactions on individual performance and species diversity have been studied extensively. However, little is known about the respective effects involving positive biological interactions, including the symbiosis between plants and ectomycorrhizal (EM) fungi. Using a greenhouse bioassay, we explored how spatial heterogeneity of natural soil inoculum influences the performance of pine seedlings and composition of their root-associated EM fungi. When the inoculum was homogenously distributed, a single EM fungal taxon dominated the roots of most pine seedlings, reducing the diversity of EM fungi at the treatment level, while substantially improving pine seedling performance. In contrast, clumped inoculum allowed the proliferation of several different EM fungi, increasing the overall EM fungal diversity. The most dominant EM fungal taxon detected in the homogeneous treatment was also a highly beneficial mutualist, implying that the trade-off between competitive ability and mutualistic capacity does not always exist.

Wild boars as spore dispersal agents of ectomycorrhizal fungi: consequences for community composition at different habitat types.

The success of dispersal events depend on the organism's ability to reach and establish in a new habitat. In symbiotic organisms, establishment also depends on the presence of their symbiont partner in the new habitat. For instance, the establishment of obligate ectomycorrhizal (EM) trees outside the forest is largely limited by the presence of EM fungi in soil. Wild boars (Sus scrofa) are important dispersal agents of EM fungal spores, particularly in the moderately dry Mediterranean region. The aim of this study was to explore how EM fungal spores dispersed by wild boars influence the EM fungal community associated with the roots of Pinus halepensis seedlings at different habitat types. Using a greenhouse bioassay, we grew pine seedlings in two soil types: old-field and forest soils mixed with either natural or autoclaved wild boar feces. In both soils, we observed a community dominated by a few EM fungal species. Geopora (85 %) and Suillus (68 %) species dominated the forest and old-field soils, respectively. The addition of natural wild boar feces increased the abundance of Tuber species in both EM fungal communities. However, this effect was more pronounced in pots with old-field soil, leading to a more even community, equally dominated by both Tuber and Suillus species. In forest soil, Geopora maintained dominance, but decreased in abundance (67 %), due to the addition of Tuber species. Our findings indicate that wild boar feces can be an important source for EM inoculum, especially in habitats poor in EM fungi such as old-fields.