ITS region of leaf metagenome: Theobroma cacao
Total genomic DNA from each subsample was extracted as described in URen & Arnold (2017). We used a designated PCR area to restrict contact with pre-PCR materials (Oita et al., 2021). Used primers for the fungal ITSrDNA region, ITS1f (5-CTTGGTCATTTAGAGGAAGTAA-3) and ITS4 (5-TCCTCCGCTTATTGATATGC-3) with modified universal consensus sequences CS1 and CS2 and 05 bp for phase-shifting. Every sample was amplified in two parallel reactions containing 1-2 L of DNA template (URen & Arnold, 2017; see also Tellez et al., 2022).