Abstract: Background and Objective: In vitro culture of Gynura procumbens adventitious roots has been carried out by various methods, sucrose concentrations and elicitation. However, adventitious root yields and flavonoid content have not yet been maximized. This study was aimed of this study was to enhance of biomass and flavonoid production from G. procumbens adventitious roots influenced by salt and nitrogen supply in Balloon Type Bubble Bioreactor (BTBB). Materials and Methods: Adventitious-root explants were obtained from leaves and were grown in MS medium supplemented with 5 mg L–1 of IBA. Then, 2 g of adventitious roots were cultured in a BTBB containing MS medium of one of four different salt strengths (½×, ¾×, 1× and 2×MS) or one of six ammonium:nitrate ratio treatments (0:30, 10:20, 15:15, 20:10, 30:0 and normal MS medium). Results: The highest adventitious root biomass yield was obtained in ½× strength MS medium and an ammonium nitrate ratio of 10:20, whereas the highest flavonoid content was obtained in 2× strength MS and an ammonium nitrate ratio of 0:30. Adventitious root biomass showed a maximum 18-fold increase compared to the initial inoculum and flavonoid production showed a 1.6-fold increase compared to the roots of the mother plant. Conclusion: These results can be employed as a basis for developing large-scale cultures because they provide information on the economical use of media and reduced application of ammonium nitrate.