Establishment of callus and cell suspensions cultures of dalbergia congestiflora (Fabaceae) to (+)-medicarpin production

Abstract

Background and Objective: Dalbergia congestiflora Pittier heartwood extracts have shown cytotoxic and antifungal activity. The medicarpin obtained of heartwood has antifungal and cytotoxic response against some diseases. This study aimed to establish the callus and cell suspensions cultures of D. congestiflora, as well as the extraction and identification of medicarpin. Materials and Methods: Leaf explants from in vitro shoots were cultured on Murashige and Skoog medium (MS) supplemented with different concentrations of benzylaminopurine (BAP) and Naphthalene Acetic Acid (NAA) to induce callus formation. The cell suspensions were established on liquid medium optimum to callus growth, inoculating 0.5 g fresh weight and agitated on orbital shaker with subcultures each 16 days. Fresh, dry weight and (+)-Medicarpin were determined at 30 days of culture in callus and every four days until day 24 in cell suspensions. (+)-Medicarpin obtained from callus and cell suspension was identifying and quantified by1HNMR mass spectroscopy analyses. Results: The optimum media to callus induction was obtained with the combination of 1.5 mg L-1 BAP and 0.1 mg L-1 NAA and was used to establish the cell suspension cultures. The growth kinetics revealed a maximum cell grows that day 16 and 80% cell viability. Both callus and cell suspensions produced (+)-Medicarpin, obtaining 3.56 mg (+)-Medicarpin/g FW at day 14 in cell suspensions. Conclusion: The results of this study demonstrate the viability to establish the callus and cell suspension cultures of D. congestiflora. The both types of cells produce (+)-Medicarpin, which is the same compound founded in heartwood of D. congestiflora.