Short-term storage of Solanum trilobatum L. synthetic seeds and evaluation of genetic homogeneity using SCoT markers

The current study demonstrates an effective protocol for encapsulation of in vitro nodal segments of Solanum trilobatum L. for short-term storage. In a modified Murashige and Skoog medium supplemented with 8.8 µM 6-benzyl adenine (BA), 0.5 µM indole-3-acetic acid (IAA), and 25 mM ammonium nitrate, a gelling matrix of 3% sodium alginate and 80 mM CaCl2.2H2O promoted the highest shoot regeneration frequency (98.4%). The effect of storage temperature, sucrose concentration, and storage time interval on shoot regrowth efficiency was assessed. Synthetic seeds maintained at culture room temperature (25 °C) were shown to be more competent than low-temperature storage (8 °C and 4 °C), with better regeneration efficiency (82.2%). The alginate matrix added with 1.5% sucrose increased the storage potential of synthetic seeds up to 12 weeks with a regeneration frequency of 57.7%. Using start codon-targeted polymorphic (SCoT) markers, the genetic fidelity of plants regrown after 12 weeks of storage was assessed. The SCoT fingerprinting approach has confirmed the post-storage genetic stability of recovered plants with their in vitro source. The present investigation is the first report on the development of synthetic seeds of S. trilobatum L., which would greatly facilitate germplasm conservation and exchange of this invaluable medicinal plant.