Before in vitro propagated Melia volkensii plants can be used for mass planting, the transition phase to in vivo conditions needs to be better controlled because too many plants are lost during acclimatization and in the field. Two experiments were set up to evaluate the effects of biological agents on the establishment of M. volkensii in vitro plantlets. The biological agents consisted of Trichotech^®, Bio-cure B^®, Rhizatech^®, Bacillus subtilis, a Trichoderma isolate and self-isolated native arbuscular mycorrhizal fungi (AMF). Regarding the latter, in soil from the nursery, the number of AMF spores increased from six spores to 400 per 100 g of soil using a trap culture, in which thirteen AMF morphotypes were identified and root colonization assessed through observation of hyphae, vesicles, coils and appressoria. The first experiment was set up in the greenhouse to investigate the efficacy of the biological agents on the hardening off. In the second, a field experiment was set up to study their effect on the early establishment of the plantlets in the field compared to seedlings. All biological agents significantly (p ≤ 0.05) improved in vitro plant survival and growth compared to the control. The highest plant height and number of leaves per plant were recorded in plants treated with Rhizatech^®, Native AMF, Bio-cure B^® and Trichoderma isolate. The treatments with Rhizatech^®, Bio-cure B^® and native mycorrhiza recorded a significantly wider stem. The root diameter of the plants treated with Rhizatech^® and Bio-cure B^® was the largest, but the plants inoculated with the native AMF had the longest roots. Moreover, the inoculated plants generally developed multiple secondary roots. After two months, AMF had clearly colonized the acclimatized plantlets. In the field experiment, the biologicals made no difference in survival rate but did produce a significantly larger leaf area after two months, with the largest leaves recorded with Rhizatech^®, native AMF and Trichotech^®. They also increased the quality index of the plants from 0.21 to 0.52. The performance of in vitro grown M. volkensii plants six months after planting in semi-arid conditions in Kiambere was better than that of seedlings. Inoculation of plants increased plant height and diameter. Thus, inoculation of biological agents is an efficient approach for improving the early growth of in vitro propagated M. volkensii plants.