This research objective was to study on shoot multiplication and microrhizome induction of Zingiber ottensii Valeton conducted from aseptic shoots approximately 2–3 cm long. The explants were cultured on MS media containing 0, 4.4, 8.8, 13.3, 17.8 and 22.2 μM6-Benzyladenine (BA) with 30 g/L sucrose for 6 weeks. The results showed that there was a statistically significant difference in the concentration of  BA affecting the average number of shoots and length of shoots. Medium supplemented with 13.3 μM BA provided the highest average number of shoots (2.8 ± 0.36 shoots/explant). Microrhizomes were induced at the base of the in vitro derived shoots upon transfer to MS medium containing various concentrations of sucrose (30, 80, 90 and 100 g/L) and grown under 16-h photoperiod or in the dark for 16 weeks. It was found that sucrose concentration and light duration affected microrhizome formation. The basal part of the shoots started swelling within 7 weeks-old culture, which was a signal for a process of rhizomes formation. At 100 g/L sucroseunder 16-h photoperiod provided the highest average number of microrhizome (3.7± 0.26). Moreover, the effect of BA at 0, 13.3, 26.6 and 39.6 μM incombination with 0, 80, 90 and 100 g/L sucrose under 16-h photoperiod on microrhizome induction was investigated. The result showed that the number of shoot, microrhizome and root were increased by using sucrose together with BA. The MS medium supplemented with 26.6 μM BA and 100 g/L sucrose were the best conditions for microrhizome induction (5.90 ±0.58).