The disclosure relates to a process of tissue culture for regeneration of Banana plants. The process consists of following steps: (a) cutting and chopping the banana suckers to obtain explants (b) sterilizing the explants by treating with at least one sterilizing agent to obtain sterilized explants (c) culturing the sterilized explants in a pre-sterilized first medium for shoot induction and multiplication to develop several proliferating shoots (d) harvesting the developed shoots (e) culturing the harvested shoots in pre-sterilized second medium for root induction and development to generate functional roots obtaining the regenerated plantlets and (f) transferring the regenerated plantlets to the soil for growing Banana plant. The disclosure also provides a culture medium, a First culture medium capable of efficient shoot induction and multiplication in explants from Banana plant and a second culture medium capable of root induction and development of rooting system.
