Rapid molecular approach for a reliable identification of Tuber spp. ectomycorrhizae

Abstract

Protocols for isolating and amplifying fungal DNA from ectomycorrhizae, overcoming the presence of PCR inhibitors, were established to ensure a fast and reliable truffle mycorrhizal identification. By using species-specific ITS primers in multiplex PCR, morphologically very similar <it>Tuber melanosporum</it>, <it>Tuber indicum</it> and <it>Tuber brumale</it> ectomycorrhizae were identified in a single step of amplification, irrespective of their physiological stage. The molecular method assures a more accurate and reliable monitoring of nursery-inoculated plants both before and after their in-field planting, than any morphological or ITS/RFLP analyses so far performed. It should also prevent frauds, already a problem, and the potential for environmental damage due to the incorrect typing of the allochthonous species <it>T. indicum</it> as European truffle species, when the identity of ectomycorrhizae is based solely on morphology.