Abstract
Ormocarpum sennoides (bone-knit) is a vulnerable medicinal plant species belongs to the family Leguminosae. The aim of the present study was to evaluate the influence of plant growth regulators (PGRs), additives (ascorbic acid, citric acid, adenine sulfate, and L-arginine), gelling agents (agar and phytagel™) and type of culture vessels on the quantitative and qualitative proliferation of shoots and the micro-morpho-anatomical state of the in vitro multiplied O. sennoides. The pre-treatment of single-node explants with antioxidants (citric acid and ascorbic acid) reduced phenolic exudates, and 93.8% bud breaking was observed on Murashige and Skoog’s (MS) medium with 1.5 mg L−1 6-benzylaminopurine (BAP). The highest rate of shoot enhancement (mean number of shoots) per explant (42.0 shoots with 6.3 cm length) was obtained on MS medium augmented with 0.5 mg L−1 each of BAP and indole-3 acetic acid (IAA) with additives. Defoliation was observed in agar-gelled medium whereas; thick and sturdy shoots with healthy leaves were detected on phytagelled MS medium. The liquid medium was found unsuitable for shoot proliferation as the shoots experienced defoliation and hyperhydricity in cultures. The micro-shoots were rooted on half strength MS medium supplemented with 2.0 mg L−1 indole-3 butyric acid (IBA) and hardened in a greenhouse. The leaves and shoots showed some characteristic anomalies in the cuticular membrane, epidermis, stomata, vein structures, chlorophyll development, hypertrophy in cortical cells, and lignifications in vascular elements when cultured in bottles capped with screw-caps. Such anomalies were absent with the shoots cultured in conical flasks capped with cotton plugs. The micro-morpho-anatomical evidences supported that MS medium gelled with phytagel™ containing additives with optimized PGRs in conical flasks could be the best physio-chemical parameters for qualitative and quantitative development of plantlets for the in vitro conservation of this vulnerable plant species.
Published Version
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