ITS rRNA ampilcon sequencing of rhizosphere mycobiome:  resident Dactylis glomerata
The total microbial genomic DNA from rhizosphere soils (filters and coarse particles) were extracted by using the method of Lueders et al. (2004), modified by Wst et al. (2016) for soils in the Biodiversity Exploratories. Frozen filter samples were cut into pieces (sterile pair of scissors) and then transferred together with the coarser soil particles to a 2 ml screw cap tube containing 0.7 g of sterilized zirconium/silica beads (0.1 mm diameter). Subsequently 750 l 120 mM sodium phosphate solution and 250 l TNS-buffer were added to the tubes. The samples were shaken with a TissueLyser II (Qiagen GmbH, Hilden, Germany; at 28 Hz for 4 min). Then continuing with the protocol. ITS rDNA amplicon libraries ITS2 region performed on the Illumina MiSeq platform (MiSeq Reagents kit v3) using the primers fITS7-ITS4 fungi.