Miroestrol and deoxymiroestrol are highly active phytoestrogens derived from the tuberous roots of Pueraria candollei var. mirifica. To date, there have been no reports regarding the production of miroestrol and deoxymiroestrol in in vitro cell culture. In this study, callus and cell suspension cultures were established for the purpose of investigating miroestrol and deoxymiroestrol content in P. candollei var. mirifica cells. Stem-derived callus cultured on Murashige and Skoog (MS) medium supplemented with 0.1 mg l⁻¹ thidiazuron (TDZ), 0.5 mg l⁻¹ naphthaleneacetic acid (NAA), and 1.0 mg l⁻¹ benzyladenine (BA) provided optimal conditions for the accumulation of deoxymiroestrol and total isoflavonoids. The calli produced 184.83 ± 20.09 µg g⁻¹ dry weight of total chromene and 20.72 ± 2.38 mg g⁻¹ dry weight of total isoflavonoid. This is the first report to suggest that callus culture is a suitable alternative method for producing miroestrol and deoxymiroestrol. Carbon sources were evaluated for the cell suspension cultures of P. candollei var. mirifica. Sucrose provided optimal conditions for biomass production, whereas fructose was the most suitable carbon source for deoxymiroestrol and isoflavonoid production. The information from our study can be employed for enhancing the production of miroestrol, deoxymiroestrol, and total isoflavonoids using in vitro cell culture of P. candollei var. mirifica.
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