Subculture is significant for maintaining the stock of roots to determine the proliferation of roots. Subcultures are needed to avoid an accumulation of toxic in the culture medium and container. In addition, there have been no reports regarding the maintenance of the adventitious root of Gynura procumbens through the subculture process. Therefore, we carried out this study is to determine the proliferation characters of adventitious root piece expiants by observing the total ofroot branches and measure the length ofroots during continued subcultures in a solid medium. The adventitious root of G. procumbens regenerated from leaf expiant in solid MS medium supplemented with 5 mg/L indole-3-butyric acid (IBA). The root pieces were subcultured with the same medium composition. Moreover, the subculture period was performed 5 t h time with a culture interval every 3 weeks. The result has shown that the subculture period affects a total of lateral root branches. The highest mean total of branches was achieved at the 4 t h subculture period (48.0±7.58), but not significant with the 1 s t subculture period (47.2±9.00). The length of the roots was affected by a total ofroot branches. The length of the roots was affected by a total of branches. The highest total roots branch on the 4 t h subculture has been dominated by high numbers root length with a score of 1 (31±2.04). On the other hand, the 1 s t subculture has dominated by high numbers root length with a score of 3 (26±0.83). The saturated concentration of I B A in medium and specific response from roots expiant tissues might cause different root proliferation during continued subcultures. In conclusion, continued subcultures of adventitious roots on certain periods can produce separate roots proliferation characters.