| Summary: | This study aimed to assess the changes in chemical properties and fungal communities in the upper mineral soil layer in managed Scots pine (<i>Pinus sylvestris</i> L.) ecosystems. Study sites were located in the three largest <i>P. sylvestris</i> massifs in Lithuania, and six sampling sites, representing different development stages of the <i>P. sylvestris</i> forest ecosystem, were selected in each of them: mature <i>P. sylvestris</i> forest stands; clear-cuts of former <i>P. sylvestris</i> mature stand; and the <i>P. sylvestris</i> plantations aging from the 1st to 4th year. High-throughput sequencing was performed to evaluate the soil fungus community at clear-cuts, early-stage reforested sites, and mature forests in Lithuania. This study has shown that, among other chemical soil parameters, the mean concentrations of mineral nitrogen (N), total phosphorus (P), and P<sub>2</sub>O<sub>5</sub> were slightly higher in the clear-cut sites, and significantly higher in the 1st year plantations compared to the mature forests. The quality filtering after PacBio sequencing showed the presence of 60,898 high-quality fungal sequences, and 1143 fungal operational taxonomic units (OTUs). The most abundant fungal OTU in our study was <i>Archaeorhizomyces</i> sp. 5425_1. In total, 70 mycorrhizal fungal OTUs were found in the soil samples at the studied sites. The most abundant ectomycorrhizal fungus identified was <i>Amanita fulva</i> (Schaeff.) Fr. The highest amount of ectomycorrhizal fungal OTUs was found in the clear-cut sites and in the mature forests. The concentrations of mineral N and P<sub>2</sub>O<sub>5</sub> in the upper mineral soil layer did not significantly affect fungal OTUs diversity. Conversely, a relatively strong correlation was obtained between the number of mycorrhizal fungal OTUs and the concentrations of total N and soil organic carbon (SOC), as well as between the numbers of saprotrophic fungal OTUs and the concentration of magnesium ions (Mg<sup>2+</sup>).
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