Shoot tip in vitro micropropagation of banana (Musa AAA East Africa) cv.’ Uganda’ resulted in off-type plants with an altered plant and fruit development. This study was carried out to determine epigenetic mechanisms underlying the altered plant and fruit development of the in vitro derived off-type banana. The off-type banana was compared with in vitro micropropagation (MP) derived normal banana and conventionally propagated (CP) banana plants with no tissue culture history in their ancestry as controls. Plant development was estimated based on plant height, girth and number of days from planting to flowering. Fruit development was measured as the number of days from flowering to fruit maturation and senescence. Mechanisms underlying the altered plant and fruit development were determined based on global cytosine DNA methylation and 2C nuclear DNA content. Leaf cytosine DNA methylation and 2C nuclear DNA content were determined using reversed phase HPLC and flow cytometer, respectively. Results showed that the off-type banana was significantly (P < 0.05) taller and delayed to flower compared with the MP and CP derived banana. Similarly, the fruits of the off-type banana had significantly (P< 0.05) longer maturation, ripening and senescence periods than those of the true-totype fruits. The off-type derived plants had lower (P < 0.05) leaf global cytosine DNA methylation and 2C nuclear DNA amount compared with the MP and CP derived plants. These findings suggest that the altered plant and fruit development of the of-type banana are possibly under the control of reduced cytosine DNA methylation and nuclear DNA content. Further studies are required to identify specific genes which affect plant and fruit development upon undergoing demethylation.