ITS2 metagenome
The ITS2 region was amplified using universal fungal primers ITS 86F (5-GTGAATCATCGAATCTTTGAA-3) and ITS 4 (5-TCCTCCGCTTATTGATATGC-3) [19]. All polymerase chain reaction (PCR) reactions involved the KAPA HiFi HotStart ReadyMix (Sigma Aldrich, USA). To remove free primers and primer dimers from the amplicons, a washing step with magnetic beads was used. For sequencing, up to 96 li-braries were pooled using all the Nextera XT indices. The process of preparing the library, putification and sequencing was performed as described [18]. To ensure taxonomic clas-sification, a quality and filtering procedure was applied to FASTq files. For the analysis of marker gene-based microbiome sequencing data, the QIIME v.2 (Quantitative Insights into Microbial Ecology) pipeline [20], version 2023.5, was employed. The ITS2 region was extracted after the sequences were denoised using the denoise wrapper [21]. Denoising and clustering were performed with DADA2 [22]. DADA2 replaces the traditional OTU selection step in amplicon sequencing workflows, instead producing higher-resolution tables of amplicon sequence variables (ASVs) [23]. Chimeric sequences were identified and filtered using USEARCH 6.1 [24]. Moreover, a taxonomy is assigned to the sequences. Further, the query sequences (ASVs) were compared with a reference database of se-quences with known taxonomic composition. Sequences belonging to chloroplasts, mi-tochondria, and eukaryotes were removed. The database used is the UNITE Fungal ITS Database v7.2, based on a set of FASTA files from: UNITE Community (https://doi.org/10.15156/BIO/587475).