Roots of Zeyheriamontana, a species native to the savanna (Cerrado) region of central Brazil, produce lapachol, a naphthoquinone with anticancer activity. Lapachol is also the precursor of β-lapachone, a novel drug candidate for preventive and adjuvant cancer therapies. The leaves of Z. montana are a renewable source of ursolic acid and oleanoic acid, compounds known for their anticancer, antioxidant and antimicrobial properties. The potential prophylactic use of β-lapachone, as well as the medicinal properties of ursolic acid, highlights the importance of this study on Z.montana’s germplasm conservation. Multiple shoots were induced on Woody Plant media with supplemented 0.1 mg·L^-1 of thidiazuron (TDZ). Rooting was promoted on half strength WP (Woody Plant media containing 1.0 mg·L^-1 of Indolbutiric acid-IBA). Plantlet acclimatization to ex-vitro condition was done at a 70% success rate using different substrates. It was possible to store Z.montana’s elite germplasm using in vitro cultures of media containing 2% sucrose plus 4% sorbitol for six months without subcultures.