ICOM1 Abstract

AMICUCCI ANTONELLA, ISMAELA ROSSI, LUCIA POTENZA, ALESSANDRA ZAMBONELLI1 , DEBORAH AGOSTINI, FRANCESCO PALMA AND VILBERTO STOCCHI. Istituto di Chimica Biologica "Giorgio Fornaini", Universitˆ di Urbino, Via Saffi, 2 - 61029 Urbino, Italy. 1Dipartimento di Protezione e Valorizzazione Agroalimentare, Via Filippo Re, 8 - 40126 Universitˆ di Bologna, Bologna, Italy. - RFLP analysis of the ITS region for the identification of ectomycorrhizae from Tuber species. The ectomycorrhizae of two economically important white truffle species, Tuber magnatum and Tuber borchii have similar morphological features from which is difficult to obtain a correct identification. In an attempt to distinguish these two species during the symbiotic phase we combined the amplification of the internal transcribed spacer (ITS) of the ribosomal DNA with the RFLP analysis. The DNA from fruitbodies of T. magnatum and T. borchii, from leaves and roots of Tilia platyphyllos and from roots of this host plant inoculated with the two ectomycorrhizal species was amplified using the "universal primers" ITS1 and ITS4. The analysis of the ITS amplification products allowed a preliminary discrimination between the two species and showed that the lengths of the two bands from the ectomycorrhizae were the same as those obtained from the two symbionts. In contrast with what we had observed after analyzing the uncut ITS region, the restriction studies with Hinf I, Mbo I and Taq I endonucleases suggested that the mycorrhizae presumed to be of T. magnatum belonged neither to the species utilized for the mycorrhization nor to other Tuber species studied in our lab. The results of this research demonstrate that ITS amplification combined with RFLP analysis represents a simple, sensitive and reliable strategy to type ectomycorrhizae from the two different species of Tuber studied.