Optimisation of tissue culture protocol using nodal part of napier grass (Pennisetum Purpureum)

Recently, Napier grass has received attention for its trait improvement due to its large potential in livestock feed. In order to allow for molecular technique for its genetic manipulation, it is necessary that the methods for tissue culture are established specifically for the species. The main objective of this research is to identify the most effective aseptic technique to achieve the most optimum antiseptic condition for tissue culture protocol. This involved with two factors which were the concentration and the time exposure of Napier grass in sodium hypochlorite (Na0CI). In addition, the study also investigated the most effective combination of auxin and cytokinin treatment for the induction of callus. At the end of this study, it is expected that the protocol established would enable mutational breeding for Napier grass. Treatment of different concentration of sodium hypochloride at concentration of 1%, 2% and 3% were used in combination with 70% alcohol for the first experiment. In the second experiment, the most optimal concentration identified was further manipulated to identify the effective duration for soaking the explant to eliminate contamination but at the same does not cause tissue necrosis. Lastly, the results of the two experiments were used to establish aseptic condition to identify the effective hormone concentration to induce callus from the Napier grass explant. The result of this project showed that the treatment of 2% Na0C1 with the time exposure of 20 minutes was the most effective to eliminate contamination. While the aim of this project is to induce callus, there were no callus formation observed at the end of this study but the survived explant instead developed shoot for explant that were treated with 2mg/L of NAA in combination with 1.5mg/L of BAP. Despite achieving the aseptic technique in the first and the second experiments, the result was inconsistent and the occurrence of contamination was still observed subsequently. Further research is required especially to use the in vitro tissue for the callus induction to eliminate contamination and to identify the effective hormone combination for callus induction instead of shoot regeneration.