Micropropagation system and their genetic fidelity evaluation from regenerated plants by ISSR and DAMD markers of Tabernaemontana alternifolia L., an endangered medicinal woody species

An important, endemic, and imperilled tropical medicinal plant called Tabernaemontana alternifolia is successfully micropropagated in this article. On Murashige and Skoog's media that had thidiazuron (TDZ; 2.5, 5.0, 7.5, and 10 mu M) supplied, high-frequency indirect shoot regeneration was induced from the callus produced from internode explants. After 12 weeks of culture, 10 mu M TDZ produced the greatest number of shoots among the different TDZ concentrations examined (15.33 shoots). From the original callus, up to three subcultures could regenerate shoots. On full and half-strength MS media containing indole-3-butyric acid (IBA) after 6 weeks of culture, rooting of the shoots was seen. Rooting of shoots was facilitated by a half-strength MS medium supplemented with 5 mu M IBA. After successfully being transplanted into a potting mixture including vermiculite and perlite and being raised there for 4 weeks, the plants were moved to one containing soil, sand, and farmyard manure and kept in the greenhouse. Inter simple sequence repeat (ISSR) and directed amplification of minisatellite DNA (DAMD) primer assays were used to assess the genetic stability of Tabernaemontana alternifolia micropropagated plants. When randomly chosen regenerated plants were compared to donor plants, a homogenous amplification profile was found, supporting the clonal fidelity of micropropagated plants. Morphometric analysis of regenerated plants was comparable with that of mother plants suggesting the genetic fidelity of regenerated plants.