American Journal of Plant Sciences, 2013, 4, 519-523 http://dx.doi.org/10.4236/ajps.2013.43066 Published Online March 2013 (http://www.scirp.org/journal/ajps) 519 In Vitro Propagation and Conservation of Zeyheria montana Mart: An Endangered Medicinal Plant Bianca Waléria Bertoni 1 , Rita Maria Moraes 2 , Laura Lemo Previdelli 1 , Paulo Sérgio Pereira 1 , Suzelei de Castro França 1 , Ana Maria Soares Pereira 1* 1 UNAERP—Unidade de Biotecnologia de Plantas Medicinais, Ribeirão Preto, Brasil; 2 National Center for Natural Products Research, The University of Mississippi, Oxford, USA. Email: * apereira@unaerp.br Received January 3 rd , 2013; revised February 5 th , 2013; accepted February 14 th , 2013 ABSTRACT Roots of Zeyheria montana, 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 pre- ventive 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 thidi- azuron (TDZ). Rooting was promoted on half strength WP (Woody Plant) media containing 1.0 mg·L −1 of indole-3- butyric acid (IBA). Plantlet acclimatization to ex-vitro condition was done at a 70% success rate using different sub- strates. 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. Keywords: Cerrado; Bignoniaceae; Micropropagation; Germplasm Storage; Lapachol and Triterpenes 1. Introduction The medicinal importance of Zeyheria montana Mart. (bolsa de pastor) a plant native to Brazilian Cerrado, ex- tends beyond the phytotherapeutic use. Its leaves produce ursolic, oleanolic and betulic acids, pentacyclictriterpe- noids, with validated biological properties, such as anti- cancer activity [1], inhibitory effects on NO production and iNOS induction [2], anti-inflammatory [3], antifun- gal [4], diuretic [5]. Ethanol extract from Z. montana leaf possesses anti-nociceptive [6] and anti-inflammatory [7] activities and flavanones showed cytotoxicity against hu- man NCI-ADR/RES and K562 cell lines [8]. Additionally, Z. montana roots are an alternative source of lapachol, a naphtoquinone with anticancer activity [9], producing 19 fold more than Tabebuia, the better known natural source of lapachol and -lapachone [10]. In the 1970s, the clinical efficacy of lapachol and related com- pounds in cancer therapy was marked, but Cragg and Newman [11] reported that the National Cancer Institute discontinued the clinical trials on lapachol and related compounds due to high toxicity. Recent findings, how- ever, indicate that -lapachone may halt the transforma- tion of normal cells to cancer cells, thus holding promise for a dual use, as an anticancer and anti-carcinogenic agent [12]. Only few conservation studies have been carried out so far as a relief effort to conserve Z. monatana germ- plasm [13-15]. Little is known on asexual propagation of elite plants to serve as propagule to achieve potential crops, and to improve quality, safety and efficacy of phy- tomedicine. According to Felippe and Silva [16], seed production is inconsistent, and flowering and fruit pro- duction are delayed for longer periods of time. Over- coming juvenility, Z. montana fruits are produced every other year. In light of the therapeutic importance of Z. montana and its endangered status, the objective of this study was to establish in vitro propagation and conservation proto- cols to preserve the species germplasm. 2. Material and Methods 2.1. Plant Material Plants and seeds were collected in Pedregulho County, São Paulo, Brazil (Lat. 20˚S 17'06", Long. 47˚W 26'12", Alt. 968 m) and the following studies were conducted micropropagation, in vitro conservation protocols and phytochemical analysis. A voucher specimen was depos- * Corresponding author. Copyright © 2013 SciRes. AJPS