Trichoderma harzianum strain T-22 (T22) is one of the most effective strains of this fungus that is able to colonise the roots of most plant species across a wide range of soil types. This fungus is used as a biocontrol agent during crop production, and for the improvement of the rooting and acclimatisation phases in plant nurseries. In vitro-cultured shoots of GiSeLa6 ® (Prunus cerasus P. canescens) and of GF677 (P. amygdalus P. persica), two important Prunus varieties used as commercial rootstocks, were inoculated with T22. The results showed that early inoculation of the fungus (at the stage of shoot transfer to root-inducing medium) seriously damaged both GiSeLa6 ® and GF677 plants; whereas, following later inoculation (7 d after shoot transfer to root-inducing medium), the plants survived and showed significant increases in shoot growth and root development. In particular, root lengths in GiSeLa6 ® and GF677 plants increased by 180% and 136%, respectively, compared to non-inoculated controls. Microscopic analysis revealed T22 hyphae spreading on the root surface in GiSeLa6 ® (fungus colonisation frequency = 20%), but not in GF677 roots. Our results demonstrate that the application of T22 during the rooting phase resulted in greater shoot lengths, as well as increased numbers of leaves, roots, and stem diameters. These morphological characteristics could increase the quality and viability of nursery planting material and provide advantages during the plant acclimatisation phase. T richoderma spp. are among the most abundant,