Timing of evolutionary innovation: independent origins of ectomycorrhizal symbiosis and adaptive radiation in a species-rich fungal clade

Short fragments of the 96 single-copy genes were amplified for 92 taxa of a basidiomycetous order Boletales. The amplicon libraries were sequenced by 2 ? 300 bp paired-end sequencing on the MiSeq platform using MiSeq v3 Reagent Kit. Thereafter, we reconstructed the origins of ectomycorrhizal (ECM) symbiosis in Boletales based on the phylogenetic tree inferred from the nucleotide sequences of 87 single-copy genes.