Regeneration and Agrobacterium-mediated genetic transformation of Terminalia bellerica Roxb.: a multipurpose tree species

An efficient in vitro transformation and plant regeneration protocol was developed for Terminalia bellerica using cotyledonary node cultures. High-frequency shoot bud proliferation was obtained on medium with 6-benzyladenine. Significant improvements in plant regeneration occurred using elevated levels of CuSO4 and CoCl2. Rooting occurred on a half-strength Murashige and Skoog medium containing indole-3-butyric acid. The rooted plants were acclimatized and transferred to field conditions. The genetic fidelity of the regenerated plants was confirmed using randomly amplified polymorphic DNA analysis. An Agrobacterium-mediated genetic transformation protocol was developed for Terminalia by varying several factors which influence T-DNA delivery. Southern blot analysis of regenerated plants confirmed selectable marker gene integration in transgenic plants. This transformation protocol can be utilized for further genetic manipulation of T. bellerica.