EFFECT OF HILUM POSITION IN THE IN VITRO REGENERATION OF MIRACLE BERRY PLANT (SYNSEPALUM DULCIFICUM) WITH 0.1BAP MG/L MS MEDIUM SUPPLEMENT

Plant tissue culture represented an essential technique in modern plant biotechnology for the propagation, conservation, and genetic improvement of valuable plant species. This study examined the in vitro regeneration response of Synsepalum dulcificum (miracle berry), a tropical West African shrub renowned for its taste-modifying glycoprotein, miraculin, under different sterilization protocols and hilum orientations. Mature seeds of Synsepalum dulcificum were obtained from the Institute of Agricultural Research and Training (IAR&T), Moor Plantation, Ibadan, and Eweje Farm Institute, Odeda and subjected to varying sterilization treatments comprising ethanol, sodium hypochlorite, and Plant Preservative Mixture (PPM) to achieve aseptic culture conditions. The findings showed the full strength MS medium supplemented with 0.1 BAP Mg/L promotes plantlet regeneration and callogenesis. The findings also revealed that the most effective sterilization treatments were the double sterilization protocols designated as DS1 and DS2. DS1 consisted of 70% ethanol for 30 seconds, followed by 20% sodium hypochlorite for 20 minutes and 10% sodium hypochlorite for 10 minutes with two drops of Tween 20, while DS2 consisted of 70% ethanol for 60 seconds, 40% sodium hypochlorite for 20 minutes, and 5% sodium hypochlorite for 5 minutes with two drops of Tween 20. The study concluded that both sterilization protocol and hilum orientation significantly influenced the regeneration success of Synsepalum dulcificum in vitro regeneration. The study recommended that double sterilization protocols should be adopted as the most effective method for establishing aseptic seed explants of miracle berry.