Abstract: Background and Objective: The most commonly used direct method in breeding triploid banana is crossing tetraploids with diploids. “Pisang Klutuk Sukun” (Musa balbisiana Colla, BB) has been associated with improved vigor and tolerance to biotic and abiotic stresses and therefore, it is a target for Musa breeding program. The objective of the study was to produce banana autotetraploid plants from diploid “Pisang Klutuk Sukun” and to evaluate the morphological characters and the reproductive potential of the autotetraploid plants. Materials and Methods: Induction of the autotetraploid was conducted using in vitro-cultured shoots of diploid “Pisang Klutuk Sukun” treated with oryzalin at a concentration of 60 μM for 7 days in a liquid MS basal medium with addition of 2 mg L–1 BA. The morphology characters of the autotetraploids were evaluated which based on 52 characters of UPOV for two reproductive cycles. The reproductive potential of the autotetraploid plants was conducted by crossing the autotetraploids with a diploid banana cultivar; “Pisang Rejang” to produce triploids. Results: The induction of autotetraploid experiment showed that 12 plants out of 34 plants of oryzalin-treated “Pisang Klutuk Sukun” were autotetraploids. The autotetraploid plants showed drooping leaves instead of erect leaves as in its diploid plants. They had fewer suckers compared to its diploid plants. The fruit diameter (width) of the autoteraploid was larger than those of its diploid. The autotetraploid “Pisang Klutuk Sukun” was successfully crossed with diploid banana cultivar, ‘Pisang Rejang’ and produced triploid hybrids. Conclusion: This study concluded that the banana autotetraploid plants of “Pisang Klutuk Sukun” were produced at a concentration of 60 μM for 7 days in a liquid MS basal medium with addition of 2 mg L–1 BA. The autotetraploids were fertile as well.