Encapsulation of Brunfelsia pauciflora microprogated shoots as a suitable conservation protocol

Abstract

The current study aimed to establish an efficient in vitro propagation and conservation protocol for Brunfelsia pauciflora plant. The effect of plant growth regulators types (BAP, Kin, 2ip and NAA) and concentrations at 0.0, 1.0, and 2.0 mg/l were studied during establishment phase in comparing with hormonal-free medium. In vitro proliferated shootlets were encapsulated using various levels of sodium alginate at 1, 2 and 3% (W/V) and calcium chloride at 2.5,5 and 10 %. Regrowth ability of encapsulated explants was evaluated after conservation for 3, 5, and 7 weeks and compared with non-encapsulated buds. The findings of the current study proved that during establishment phase, applying BAP at 2.0 mg/l gave the best results during the three successive subcultures. The best treatment 2ip (2.0mg/l) + BAP (2.0mg/l) + NAA (2.0mg/l) that was considered ideal propagation protocol for shooting and rooting of Brunfelsia. Successful plant recovery from encapsulated explants using low concentrations of sodium alginate (1%) and calcium chloride at 2.5%, proved that the explants could to be survive at highest percent (100%) during conservation periods 3, 5 and 7weeks. Increasing calcium chloride to 5% with the same concentration of sodium alginate (1%) caused gradually reduced percent of survival to 53.33% with increasing conservations period (from 3 to 7weeks). It is the first paper of the application of syndetic seed technique on Brunfelsia for the shortest period conservation and successive renovation of shootlets whose genetic stabilization was evaluated by analysis of RAPD that proved that the genetic similarity ranged from 56.1 to 80%. These results were confirmed by anatomical sections made in Brunfelsia leaf blade.