High frequency in vitro plantlet regeneration in Solanum trilobatum L., an important ethno-medicinal plant and confirmation of genetic fidelity of R1 plantlets by using ISSR and RAPD markers

An efficient and reproducible protocol was developed for in vitro clonal propagation of Solanum trilobatum L., an ethno-medicinally important plant. Leaf, stem and cotyledon explants were used for callus induction and shoot regeneration via indirect organogenesis. Initially, maximum amounts (mg) of green friable callus (312.86 ± 0.50, 285.3 ± 0.40 and 305.13 ± 0.62) was induced from leaf, stem and cotyledon explants, respectively, on Murashige and Skoog (MS) medium supplemented with 13.57 μM L−1 of 2,4-dichloro phenoxy acetic acid (2,4-D) and 2.21 μM L−1 of 6-benzylaminopurine (BAP), after 4 weeks of culture. Highest mean number of shoots (69.2 ± 0.73, 34.1 ± 0.62 and 57.1 ± 0.62) were differentiated de novo from leaf, stem and cotyledon calluses, respectively, when cultured on MS medium amended with 2.27 μM L−1 of Thidiazuron (TDZ) and 2.68 μM L−1 of Naphthaleneacetic acid (NAA), after 4 weeks of culture. Maximum rooting response (97%) with mean number of roots (31.7 ± 0.61 roots per shoot) was observed from shoots when cultured on half strength MS medium supplemented with 4.90 μM L−1 of indole-3-butyric acid (IBA), after 4 weeks of culture. In vitro raised plantlets (R1) of S. trilobatum were hardened in plastic pots, acclimatized in green house and successfully transferred to field conditions with 82% survivability. ISSR and RAPD based PCR banding profile of the acclimated R1 plantlets was confirmed as synonymous to the mother plant. © 2019, Society for Plant Research.