High incidence regeneration system in Ceratonia siliqua L. articulated with SEM and biochemical analysis during developmental stages

Abstract

The article describes refined regeneration process in Ceratonia siliqua using different plant growth regulators along with antioxidant, SEM and IRGA analysis to understand the developmental behaviour of the plantlets. The present study describes a simplified seed germination process in Ceratonia siliqua L. and gives a comparative analysis of hormonal supply for enhanced adventitious shoot regeneration through aseptic seedling-derived cotyledonary nodes. The axillary bud induction and multiplication was greatly influenced by the concentration and type of cytokinin viz. 6-benzylaminopurine (BA), meta-Topolin (mT), Kinetin (Kn) and Thiadiazuron (TDZ). Adenine-based cytokinins, BA and mT, augmented in Murashige and Skoog medium, were proved to be more responsive. The medium containing 10 µM BA produced a maximum of 9.1 ± 3.0 shoots/ explant. A combination of optimal concentration of BA with NAA (0.5 µM) however, resulted into a significant production of 28.10 ± 0.14 shoots per CN. Half-strength MS + IBA (10 µM) in the presence of light ensured more number of root formation while half-strength MS + IBA (5.0 µM) and stimulation in the dark for 1 week ensured better root growth (4.98 ± 0.08 cm). Plantlets were successfully acclimatized in soilrite™, showing 65% survival. The work is supported with the studies based on antioxidant enzyme activity as well as on net photosynthetic rate and its related attributes in comprehension with SEM analysis of leaves showing stomatal micromorphology during three stages: in vitro, during acclimatization and in net house conditions. The studies during acclimatization reflect better understanding of the stabilization of micropropagated plantlets to the environmental conditions.