Direct organogenesis, physiochemical evaluation, chemical characterization and genetic uniformity analysis through SCoT marker in Celastrus paniculatus (Willd.): A medicinally important plant

Celastrus paniculatus (Willd.) is a rare woody climbing shrub with high medicinal value used in primary healthcare system and herbal drug formulation as its various parts have immense medicinal properties. The current study focuses to design a refined methodology for the micropropagation of C. paniculatus using young nodal segment explants. A reliable shoot multiplication protocol was attained on Murashige and Skoog (MS) medium augmented with 6-benzylaminopurine (BAP) (5.0 mu M) + meta-Topolin (mT) (2.5 mu M) + gibberellic acid (GA3) (2.5 mu M) + adenine sulphate (ADS) (20.0 mu M) which gave a maximum number of 26.0 +/- 0.01 shoots/ explant along with 8.5 +/- 0.02 cm of shoot length. In vitro regenerated microshoots were accomplished to ex vitro rooting efficiently with a 15 min pulse treatment with two ethylene inhibitors [salicylic acid (SAA) and silver nitrate (AgNO3)] and then transfer to soilriteTM. The treatment of SAA (200 mu M) proved better for healthy root formation with 10.5 +/- 0.017 roots/microshoot and a mean root length of 3.5 +/- 0.024 cm under culture condition. The genetic stability of in vitro regenerated plantlets and mother plant was validated by start codon targeted (SCoT) primer based molecular analysis. Healthy adaptation of these plantlets to natural conditions was also revealed by numerous biochemical and photosynthetic parameters along with Scanning Electron Microscopic (SEM) analysis of leaves. Furthermore, a comparison of secondary metabolites from mother and regenerated plants was done by Gas Chromatography and Mass Spectrometry (GC-MS) analysis.