Synthesis of nonembryonic synseed, short term conservation, phytochemical evaluation and assessment of genetic stability through SCoT markers in Decalepis salicifolia

Abstract

The current work specifies an effort to conserve the highly endangered plant Decalepis salicifolia by utilizing encapsulation technology for high plantlet recovery, short-term storage, and conservation. In order to determine the best culture conditions for maximizing the ability of synseeds to develop into complete plantlets, nodal segments (NS) were encapsulated in a sodium alginate (SA) matrix. The best solution for creating isodiametric beads was determined to be a 3% sodium alginate with 100 mM CaCl₂ solution. Regeneration of the encapsulated segment was reported directly or after storage at 4 °C for up to 8 weeks. The highest shoot regrowth frequency (90.8%) and highest microshoots per encapsulated nodal segment i.e. 4.90 were recorded when Murashige and Skoog (MS) basal medium was supplemented with 5.0 µM 6-benzyl adenine (BA) + 1.0 µM indole-3-butyric acid (IBA) and encapsulated nodal segment were inoculated onto the nutrient medium comprised of MS + 5.0 µM BA. Microshoots rooted effectively on MS medium augmented with indole-3-butyric acid IBA (2.5 µM). Plantlets achieved from preserved synthetic seeds were acclimatized and relocated in the natural condition successfully with an immortality rate of 87.1%. Subsequently, it was planted in garden soil and exhibited no morphological changes. Gas Chromatography and Mass Spectrometry (GC-MS) of leaf extract, obtained from the donor plant as well as in vitro derived encapsulated regenerated plantlets shows the presence of diverse chemical compounds of immense pharmacological properties and number of biochemical compounds are almost similar in both, which established the biochemical similarity between them. Genetic similarity between the donar plant and the synseed-derived plant was confirmed by the presence of monomorphic bands produced with the help of the start codon targeted (SCoT) marker.