Key message Shoots of B. vulgaris cultured in TIS showed better morpho-physiological and biochemical characteristics, without hyperhydricity during in vitro multiplication.The aim of this study was to compare two in vitro culture methods: the liquid culture in static and temporary immersion system (TIS) and in terms of shoot multiplication and morpho-physiological response of shoots of Bambusa vulgaris Schrad. ex Wendl. The morpho-physiological parameters such as number of shoots, shoot length and number of leaves per shoot were measured omicron n the plants cultivated in vitro. In addition, water content, total phenolic content, and lignin content were determined omicron n the shoots developed under both conditions. The results of this study show that the use of TIS favored the propagation, morphology and biochemistry of the shoots grown in TIS. In this system, the length, number of shoots and leaves per explant increased. Likewise, the highest values for phenolic and lignin content were obtained. However, shoots grown in static liquid culture medium experienced an increase in water content and hyperhydricity. Lignin and total phenolic content also decreased. This study shows that in vitro multiplication of shoots at TIS can be a successful alternative to the conventional liquid culture system, while maintaining morphological and biochemical quality for reforestation purposes.