The Mycelium Donor Plant system (MDP) was adapted to study the time course of the colonization of Pyrus communis by Rhizophagus irregularis under in vitro conditions. Isolated germinated spores did not colonize pear roots. Inoculum composed of R. irregularis spores/mycelium associated with chicory root fragments was used to inoculate Medicago truncatula which became thereafter the MDP of pear plantlets. Typical intraradical structures (hyphae, arbuscules, spores/vesicles) and appressoria were observed in the pear roots. During acclimatization, the pear plants formed a densely branched root system. R. irregularis colonization not only altered the root architecture but also changed the nutrient composition of the acclimatized pear plantlets.