Somatic embryogenesis and plant regeneration from leaf callus with genetic stability validation using SCoT markers in Paramignya trimera, a medicinal plant native to Vietnam

Xao tam phan (Paramignya trimera (Oliv.) Guillaum) is a medicinal plant native to Vietnam, that is renowned for its therapeutic properties, particularly for the treatment of various ailments, including cancer. This study investigated in vitro propagation of P. trimera through somatic embryogenesis and shoot organogenesis using leaf callus. Various culture media, plant growth regulators, malt extract, and carbon sources were evaluated to optimize callus induction and somatic embryo formation from leaf explants. DNA barcoding confirmed 96.96% to 100% homology with P. trimera specimens from Khanh Hoa province, Vietnam. The highest callus formation rate, reaching 100%, was observed in one-year-old explants cultured in Woody Plant Medium (WPM) supplemented with 2.0 mg L-1 naphthaleneacetic acid (NAA) and 0.2 mg L-1 benzylaminopurine (BAP) in the dark for over six weeks. In WPM supplemented with 30 g L-1 sucrose, 4.0 mg L-1 BAP, and 500 mg L-1 malt extract, globular stage embryos developed into embryoids and shoots and buds clumped at 10 and 18 weeks, respectively. Shoot organogenesis was observed in WPM supplemented with 30 g L-1 sucrose and 0.07 mg L-1 thidiazuron (TDZ) after 18 weeks of culture. Genetic fidelity assessments using 12 SCoT markers indicated that in vitro plantlets were homologous to the mother plant. This study provides a viable method for the conservation and sustainable cultivation of Xao tam phan, ensuring a stable supply of this valuable medicinal resource.