Ferocactus peninsulae (Cactaceae) (F.A.C. Weber) Britton & Rose is one of the most significant cacti in landscape projects and is distributed in arid climates. Conventionally, these are propagated by seeds; however, seed quantities remain insufficient for mass propagation due to florivore damage and spontaneous abortions of buds and flowers. The slow growth and the high risk of extinction of this species have increased the need for effective propagation techniques. Thus, this research aimed to produce multiple shootlets in vitro involving the areoles of barrel cactus. The in vitro germinated seedlings were excised to a length of 0.5 cm and cultivated on Murashige and Skoog (MS) nutrient medium containing various amounts of 6-benzylaminopurine (BAP) and meta-Topolin (mT). Successive micro-structural evaluation of explant tissues indicated that shootlets regenerated from areoles by forming actively dividing meristematic regions, which increase to form shoot protuberances. Numerous well-formed shootlets (5.7 shootlets with 2.5 cm length) with areoles were produced after 8 weeks of culture on 0.5 mg L^-1 mT. The basal portions of shootlets were composed of parenchymatous files of cells with a broad point of attachment to the explant tissues. Subsequent subculture produced multiple shootlets (31.0 shoots per explants per culture bottle with 4.3 cm average length) on 0.5 mg L^-1 mT + 0.15 mg L^-1 α-Naphthalene acetic acid (NAA). The positive impact of mT on increased rhizogenesis was also observed. The shootlets derived from mT (cytokinin) and NAA (auxin) medium were healthier and exhibited better rooting (93.3% response with 8.0 roots per shootlet) on ½ strength nutrient medium + 0.75 mg L^-1 IBA. The study demonstrates that areoles could be induced using mT to regenerate multiple shoots for large-scale propagation of cacti.