The species diversity of xylobiont fungi and bacteria in a catastrophic windthrow of a multi-species broad-leaved forest in the Kaluzhskie Zaseki State Nature Reserve will be assessed by means of metagenomic analysis. Forests in the Reserve are unique in their multi-species composition, high tree age and relatively large area; they are "reference areas" for both forest biodiversity protection and ecosystem-based forest management in the temperate forest region. Coarse woody debris (deadwood) plays a major role in nutrient cycling as a temporary storage stock of carbon and macro- and micronutrients,which become available to biota during the deadwood decomposition. Understanding the factors affecting wood decomposition is necessary to assess the carbon sequestration capacity of forests, which is a key issue addressing climate change. Fungi and bacteria are the main destructors of deadwood; xylotrophic fungal communities being highly diverse and including rare and endangered taxa. Despite the high importance of research on wood decomposition processes, their detailed analysis has only recently begun. It becomes possible due to the emergence of the newest methods for analyzing natural ecosystems, methods for assessing their diversity and functioning. One of these newest methods is metagenomics research, which allows analysis of fungal diversity not only by fruiting bodies, but also by their dark diversity and by the bacterial community that develops in dead wood. The novelty and originality of the research is determined both by the integrated approach (ecology of biosystem, mycology and structural genomics) and by the uniquely wide range of research objects. For the first time, metagenomics methods will be applied to identify the diversity of xylobiont fungi and bacteria on deadwood samples of eight tree species (Quercus robur, Fraxinus excelsior, Tilia cordata, Ulmus glabra, Acer platanoides, Populus tremula, Betula pendula, and Picea abies) at five decay stages; all the deadwood was formed due to the natural disturbance by catastrophic windthrow happened in 2006. A significant objective of the project is DNA barcoding of macromycetes based on sequencing of fruit bodies collected in habitats where sampling for metagenomic analysis will be carried out and for which there is no information in reference databases on nucleotide sequences. Sanger sequencing of freshly collected fruit body samples of fungi that develop directly in the habitats under study will allow the generation of an array of reference nucleotide sequences and reduce the number of 'nameless' OTUs when interpreting metagenomics sequencing results. The project will conclude with an assessment of fungal and bacterial diversity and an analysis of the relationships between the diversity and composition of xylobionts and the taxonomic identity, decomposition stage of the wood samples, and their physical and chemical characteristics as well.
