Long-term Consequences on Soil Fungal Community Structure: Monoculture Planting and Natural Regeneration.

Understanding the regeneration and succession of belowground communities, particularly in forests, is vital for maintaining ecosystem health. Despite its importance, there is limited knowledge regarding how fungal communities change over time during ecosystem development, especially under different forest restoration strategies. In this study, we focused on two restoration methods used in northern Japan: monoculture planting and natural regeneration. We examined the responses of the fungal community to monoculture plantations (active tree planting) and naturally regenerated (passive regeneration) forests over a 50-year chronosequence, using natural forests as a reference. Based on DNA metabarcoding, we assessed the richness of fungal Operational Taxonomic Units (OTUs) and their dissimilarity. Our findings revealed that soil fungal richness remained stable after natural regeneration but declined in monoculture plantations, from 354 to 247 OTUs. While the compositional dissimilarity of fungal assemblages between monoculture plantations and natural forests remained consistent regardless of the time since tree planting, it significantly decreased after natural regeneration, suggesting recovery to a state close to the reference level. Notably, the composition of key functional fungal groups-saprotrophic and ectomycorrhizal- has increasingly mirrored that of natural forests over time following passive natural regeneration. In summary, our study suggests that monoculture plantations may not be effective for long-term ecosystem function and service recovery because of their limited support for soil fungal diversity. These results underscore the importance of natural regeneration in forest restoration and management strategies.

A large-scale field experiment of artificially caused landslides with replications revealed the response of the ground-dwelling beetle community to landslides.

Precipitation-induced landslides, which are predicted to increase under the changing climate, may have large impacts on insect community properties. However, understanding of how insect community properties shift following landslides remains limited because replicated research involving landslides, which are large-scale disturbances with stochastic natural causes, is difficult. To tackle this issue, we conducted a large-scale field experiment by artificially causing landslides at multiple sites. We established 12 landslide sites, each 35 m × 35 m, and 6 undisturbed sites in both planted and natural forests and collected ground-dwelling beetles 1 year later. We found that forest type (i.e., pre-disturbance vegetation) did not affect the structure of a ground-dwelling beetle community disturbed by a landslide (landslide community), but the structure of an undisturbed community was affected by forest type. Moreover, the structures of landslide and undisturbed communities were completely different, possibly because landslides create harsh environments that act as an ecological filter. Thus, a niche-selection process may have a critical role in community assembly at landslide sites. There were no significant differences in species diversity between undisturbed and landslide communities, suggesting that landslides to not reduce species richness overall. However, among-site variability in species composition was much greater at landslide sites than at undisturbed sites. This result suggests that stochastic colonization predominated at the landslide sites more than undisturbed sites. Synthesis and applications. Overall, our results suggest that both deterministic and stochastic processes are critical in community assembly, at least in the early post-landslide stage. Our large-scale manipulative field experiment with replications has thus resulted in new insights into biological community properties after a landslide.

Environmental DNA enables detection of terrestrial mammals from forest pond water.

Terrestrial animals must have frequent contact with water to survive, implying that environmental DNA (eDNA) originating from those animals should be detectable from places containing water in terrestrial ecosystems. Aiming to detect the presence of terrestrial mammals using forest water samples, we applied a set of universal PCR primers (MiMammal, a modified version of fish universal primers) for metabarcoding mammalian eDNA. The versatility of MiMammal primers was tested in silico and by amplifying DNAs extracted from tissues. The results suggested that MiMammal primers are capable of amplifying and distinguishing a diverse group of mammalian species. In addition, analyses of water samples from zoo cages of mammals with known species composition suggested that MiMammal primers could successfully detect mammalian species from water samples in the field. Then, we performed an experiment to detect mammals from natural ecosystems by collecting five 500-ml water samples from ponds in two cool-temperate forests in Hokkaido, northern Japan. MiMammal amplicon libraries were constructed using eDNA extracted from water samples, and sequences generated by Illumina MiSeq were subjected to data processing and taxonomic assignment. We thereby detected multiple species of mammals common to the sampling areas, including deer (Cervus nippon), mouse (Mus musculus), vole (Myodes rufocanus), raccoon (Procyon lotor), rat (Rattus norvegicus) and shrew (Sorex unguiculatus). Many previous applications of the eDNA metabarcoding approach have been limited to aquatic/semiaquatic systems, but the results presented here show that the approach is also promising even for forest mammal biodiversity surveys.

An aquatic vertebrate can use amino acids from environmental water.

Conventional food-web theory assumes that nutrients from dissolved organic matter are transferred to aquatic vertebrates via long nutrient pathways involving multiple eukaryotic species as intermediary nutrient transporters. Here, using larvae of the salamander Hynobius retardatus as a model system, we provide experimental evidence of a shortcut nutrient pathway by showing that H. retardatus larvae can use dissolved amino acids for their growth without eukaryotic mediation. First, to explore which amino acids can promote larval growth, we kept individual salamander larvae in one of eight different high-concentration amino acid solutions, or in control water from which all other eukaryotic organisms had been removed. We thus identified five amino acids (lysine, threonine, serine, phenylalanine, and tyrosine) as having the potential to promote larval growth. Next, using 15N-labelled amino acid solutions, we demonstrated that nitrogen from dissolved amino acids was found in larval tissues. These results suggest that salamander larvae can take up dissolved amino acids from environmental water to use as an energy source or a growth-promoting factor. Thus, aquatic vertebrates as well as aquatic invertebrates may be able to use dissolved organic matter as a nutrient source.

Low multifunctional redundancy of soil fungal diversity at multiple scales.

Theory suggests that biodiversity might help sustain multiple ecosystem functions. To evaluate possible biodiversity-multifunctionality relationships in a natural setting, we considered different spatial scales of diversity metrics for soil fungi in the northern forests of Japan. We found that multifunctionality increased with increasing local species richness, suggesting a limited degree of multifunctional redundancy. This diversity-multifunctionality relationship was independent of the compositional uniqueness of each community. However, we still found the importance of community composition, because there was a positive correlation between community dissimilarity and multifunctional dissimilarity across the landscape. This result suggests that functional redundancy can further decrease when spatial variations in identities of both species and functions are simultaneously considered at larger spatial scales. We speculate that different scales of diversity could provide multiple levels of insurance against the loss of functioning if high-levels of local species diversity and compositional variation across locations are both maintained. Alternatively, making species assemblages depauperate may result in the loss of multifunctionality.