ABSTRACT: Mycorrhizae are mutualistic associations between fungi and plant roots. These symbiotic associations are abundant and occur in 75 to 80 % of plants. Ectomycorrhizal fungi are very important in ecosystems, because their mutualistic association with plants of different species helps nutrients and water absorption, as well as protection of the host plant against pathogens and abiotic stresses. Most ectomycorrhizal fungi belong to the Basidiomycota class, such as the following genera: Amanita, Hebeloma, Hysterangium, Laccaria, Lactarius, Rhizopogon, Russula, Scleroderma, Suillus, Tricholoma, among others. Morphological studies on ectomycorrhizae report important results in understanding the species biodiversity. However, the use of molecular biology nowadays is indispensable. Among the various molecular tools available, there is consensus about the use of tools based on sequencing of the Internal Transcribed Spacer (ITS) of fungi rDNA, aiding in species characterization and construction of phylogenetic studies. The ITS region is of easy amplification, it has multicopy nature and enables differentiation between species. The objective of this study was to show that the use of molecular biology tools associated with morphotyping to characterize species of ectomycorrhizae is more effective than when they are used on their own.