BACKGROUND: Fine woody debris (FWD; deadwood < 10 cm diameter) is a crucial but often overlooked component of forest ecosystems. It provides habitat for microbial communities and enhances soil fertility through nutrient cycling. This role is especially important in managed forests, which typically have limited deadwood stocks. Climate change is increasing forest disturbances and expanding early successional forests with low canopy cover, yet the effects on microbial communities and related processes remain poorly understood. RESULTS: In a ten-year canopy manipulation experiment, we examined the decomposition of FWD of Fagus sylvatica and Abies alba. Increased canopy openness significantly decreased bacterial diversity in decomposing FWD and altered the community composition in surrounding soil. Decomposition time was the main factor shaping bacterial community structure in FWD, with tree species and canopy cover also contributing. We identified bacterial groups involved in carbohydrate degradation, fungal biomass breakdown, and nitrogen fixation. Importantly, bacterial communities in fully decomposed FWD remained distinct from soil communities. CONCLUSIONS: Deadwood decomposition and nutrient cycling are driven by complex ecological interactions. Microbial community dynamics are influenced by the interplay of FWD decomposition stage, tree species, and microclimatic conditions. Bacterial communities, although less frequently studied in this context, appear more stable over time than previously studied fungi. This stability may help sustain decomposition processes and nutrient turnover under the environmental variability associated with global change.

• Keywords
• Bacterial community, Canopy cover, Deadwood, Decomposition, Ecology, Fine woody debris, Microclimate, Succession, Temperate forest,
• Publication type
• Journal Article MeSH

BACKGROUND: Deadwood decomposition is an essential ecological process in forest ecosystems, playing a key role in nutrient cycling and carbon sequestration by enriching soils with organic matter. This process is driven by diverse microbial communities encompassing specialized functions in breaking down organic matter, but the specific roles of individual microorganisms in this process are still not fully understood. RESULTS: Here, we characterized the deadwood microbiome in a natural mixed temperate forest in Central Europe using PacBio HiFi long-read sequencing and a genome-resolved transcriptomics approach in order to uncover key microbial contributors to wood decomposition. We obtained high quality assemblies, which allowed attribution of complex microbial functions such as nitrogen fixation to individual microbial taxa and enabled the recovery of metagenome-assembled genomes (MAGs) from both abundant and rare deadwood bacteria. We successfully assembled 69 MAGs (including 14 high-quality and 7 single-contig genomes) from 4 samples, representing most of the abundant bacterial phyla in deadwood. The MAGs exhibited a rich diversity of carbohydrate-active enzymes (CAZymes), with Myxococcota encoding the highest number of CAZymes and the full complement of enzymes required for cellulose decomposition. For the first time we observed active nitrogen fixation by Steroidobacteraceae, as well as hemicellulose degradation and chitin recycling by Patescibacteria. Furthermore, PacBio HiFi sequencing identified over 1000 biosynthetic gene clusters, highlighting a vast potential for secondary metabolite production in deadwood, particularly in Pseudomonadota and Myxococcota. CONCLUSIONS: PacBio HiFi long-read sequencing offers comprehensive insights into deadwood decomposition processes by advancing the identification of functional features involving multiple genes. It represents a robust tool for unraveling novel microbial genomes in complex ecosystems and allows the identification of key microorganisms contributing to deadwood decomposition.

• Keywords
• Biosynthetic gene cluster, Carbohydrate-active enzyme, Deadwood microbiome, Metagenome-assembled genome, Metagenomics, Microbial decomposition, Nitrogen fixation,
• Publication type
• Journal Article MeSH

Fine woody debris (FWD) represents the majority of the deadwood stock in managed forests and serves as an important biodiversity hotspot and refuge for many organisms, including deadwood fungi. Wood decomposition in forests, representing an important input of nutrients into forest soils, is mainly driven by fungal communities that undergo continuous changes during deadwood decomposition. However, while the assembly processes of fungal communities in long-lasting coarse woody debris have been repeatedly explored, similar information for the more ephemeral habitat of fine deadwood is missing. Here, we followed the fate of FWD of Fagus sylvatica and Abies alba in a Central European forest to describe the assembly and diversity patterns of fungal communities over 6 years. Importantly, the effect of microclimate on deadwood properties and fungal communities was addressed by comparing FWD decomposition in closed forests and under open canopies because the large surface-to-volume ratio of FWD makes it highly sensitive to temperature and moisture fluctuations. Indeed, fungal biomass increases and pH decreases were significantly higher in FWD under closed canopy in the initial stages of decomposition indicating higher fungal activity and hence decay processes. The assembly patterns of the fungal community were strongly affected by both tree species and microclimatic conditions. The communities in the open/closed canopies and in each tree species were different throughout the whole succession with only limited convergence in time in terms of both species and ecological guild composition. Decomposition under the open canopy was characterized by high sample-to-sample variability, showing the diversification of fungal resources. Tree species-specific fungi were detected among the abundant species mostly during the initial decomposition, whereas fungi associated with certain canopy cover treatments were present evenly during decomposition. The species diversity of forest stands and the variability in microclimatic conditions both promote the diversity of fine woody debris fungi in a forest.

• Keywords
• canopy cover, deadwood, decomposition, ecology, fungal community, microclimate, succession, temperate forest,
• Publication type
• Journal Article MeSH

Deadwood represents significant carbon (C) stock in a temperate forests. Its decomposition and C mobilization is accomplished by decomposer microorganisms - fungi and bacteria - who also supply the foodweb of commensalist microbes. Due to the ecosystem-level importance of deadwood habitat as a C and nutrient stock with significant nitrogen fixation, the deadwood microbiome composition and function are critical to understanding the microbial processes related to its decomposition. We present a comprehensive suite of data packages obtained through environmental DNA and RNA sequencing from natural deadwood. Data provide a complex picture of the composition and function of microbiome on decomposing trunks of European beech (Fagus sylvatica L.) in a natural forest. Packages include deadwood metagenomes, metatranscriptomes, sequences of total RNA, bacterial genomes resolved from metagenomic data and the 16S rRNA gene and ITS2 metabarcoding markers to characterize the bacterial and fungal communities. This project will be of use to microbiologists, environmental biologists and biogeochemists interested in the microbial processes associated with the transformation of recalcitrant plant biomass.

• MeSH
• Bacteria classification   MeSH
• Fagus microbiology   MeSH
• Wood microbiology   MeSH
• Ecosystem MeSH
• Fungi classification   MeSH
• Forests MeSH
• Metagenome * MeSH
• DNA, Ribosomal Spacer genetics   MeSH
• Microbiota * MeSH
• RNA, Ribosomal, 16S genetics   MeSH
• Trees microbiology   MeSH
• DNA Barcoding, Taxonomic MeSH
• Publication type
• Journal Article MeSH
• Dataset MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic MeSH
• Names of Substances
• DNA, Ribosomal Spacer MeSH
• RNA, Ribosomal, 16S MeSH

The bacterial genus Sodalis is represented by insect endosymbionts as well as free-living species. While the former have been studied frequently, the distribution of the latter is not yet clear. Here, we present a description of a free-living strain, Sodalis ligni sp. nov., originating from decomposing deadwood. The favored occurrence of S. ligni in deadwood is confirmed by both 16S rRNA gene distribution and metagenome data. Pangenome analysis of available Sodalis genomes shows at least three groups within the Sodalis genus: deadwood-associated strains, tsetse fly endosymbionts and endosymbionts of other insects. This differentiation is consistent in terms of the gene frequency level, genome similarity and carbohydrate-active enzyme composition of the genomes. Deadwood-associated strains contain genes for active decomposition of biopolymers of plant and fungal origin and can utilize more diverse carbon sources than their symbiotic relatives. Deadwood-associated strains, but not other Sodalis strains, have the genetic potential to fix N2, and the corresponding genes are expressed in deadwood. Nitrogenase genes are located within the genomes of Sodalis, including S. ligni, at multiple loci represented by more gene variants. We show decomposing wood to be a previously undescribed habitat of the genus Sodalis that appears to show striking ecological divergence.

• Keywords
• Sodalis, deadwood, free-living, insect symbionts, nitrogen fixation, non-symbiotic,
• Publication type
• Journal Article MeSH