title: Native litter
design: From each composite sample 2 g were macerated in liquid nitrogen and extracted using a DNeasy Plant Mini Kit (Qiagen Inc., Valencia, CA, USA). The samples were treated with 2 g of RNase (PureLink) per L of DNA sample. DNA concentration was quantified with a Qubit dsDNA BR Assay Kit (Thermo Fisher Scientific). PCR reaction and sequencing were performed by BPI Biotechnology Company. The PCR reaction produced a final volume of 20 L, containing 10 L of GoTaq Colorless Master Mix 2x (Promega, USA) 0.3 M of ITS-86F primer [25], 0.3 M of ITS-4R primer, 1 L of genomic DNA and ultrapure water. Amplification consisted of initial denaturation at 95 C for 5 min, followed by 35 cycles of denaturation at 95 C for 30 s, annealing at 56 C for 40 s, extension at 72 C for 1 min, and a final extension at 72 C for 5 min. The ITS1 was amplified in a multiplexing PCR using a set of barcoded primers (ITS-86F and ITS-4R) conducted in a Veriti Thermal Cycler thermal cycler (Applied Biosystems). The reaction was confirmed in 2% agarose gel stained with UniSafe Dye 0.03% (v/v), 400 bp (amplicon size). The amplicons were purified using an Agencourt AMPure XP magnetic bead (Beckman Coulter) and indexing reaction by Nextera XT Index kit (Illumina). The libraries were submitted to purification steps using Agencourt AMPure XP magnetic beads (Beckman Coulter) and quantification by PCR in Real Time using KAPA-KK4824 Library Quantification Kit (Illumina/Universal) and sequenced using the Illumina MiSeq new generation sequencing system (Illumina Sequencing) with MiSeq Reagent Kit V2 Nano 500 cycles - 2 x 250 bp reading.