Bamboo species—the only herbaceous trees—have unique structural and physiological characteristics that differ from those of other tree taxa. However, the role of night-time water use in bamboo is poorly understood and has rarely been investigated. We studied the day- and night-time sap flow response to culm age and rhizome structure in three age levels (juvenile, mature, and senescent) of Phyllostachys pubescens growing in the Nankun Mountain Natural Reserve, South China. We found that sap flow density and whole-tree hydraulic conductance decreased with culm age. After cutting of rhizome, the day-time sap flow and night-time water recharge decreased obviously. In addition, night-time water recharge accounted for the largest proportion (up to 30%) of total daily transpiration in normal senescent bamboos. Therefore, our study indicates that the connected rhizome system and night-time water recharge played a significant role in water compensation during the day and at night in bamboos. Night-time water recharge is especially critical to senescent bamboos, given their weaker transpiration due to the lower whole-tree hydraulic conductance, and consequently, they are more dependent on night-time water recharge for fulfilling their whole-day water consumption needs.