Cell totipotency and pluripotency are the cellular basis for root regeneration in Platycladus orientalis cuttings, and the regeneration of adventitious roots is a key determinant for improving stem-cutting. However, the cellular basis and physiological regulation of adventitious root formation are still ambiguous. In this work, root primordia initiation and organogenesis were histologically observed, dynamic alterations in soluble proteins were monitored, and Tandem Mass Tag protein profiling during adventitious root development was carried out.