In vitro-derived petiole and leaf explants of Viola cornuta L. ‛Lutea Splendens’ were cultured on half-strength Murashige and Skoog (MS) medium with several concentrations of 6-benzylaminopurin (BAP), 6-furfurylaminopurine (KIN), N-phenyl-N’(1,2,3-thidiazol-5-yl) urea (TDZ) or N-(2-chloro-4-pyridyl)-N’phenylurea (CPPU) alone or supplemented with indole-3-acetic acid (IAA) or α-naphthaleneacetic acid (NAA). The most efficient direct adventitious shoot induction (21%) without callus formation was obtained using petiole explants cultured on half-strength MS medium containing 2.02 μM CPPU. Significantly higher shoot multiplication was achieved on medium supplemented with TDZ (7.6 shoots) or CPPU (7.1 shoots) than on medium supplemented with NAA and BAP (2.9 shoots). Shoots were rooted on half-strength MS medium without plant growth regulators.