Bio-water saving is to increase water use efficiency of crops or crop yield per unit of water input. Plant water use efficiency is determined by photosynthesis and transpiration, for both of which stomata are crucial. Stomata are pores on leaf epidermis for both water and carbon dioxide fluxes that are controlled by two major factors: stomatal behavior and density. Stomatal behavior has been the focus of intensive research, while less attention has been paid to stomatal density. Recently, a number of genes controlling stomatal development have been identified. This review summarizes the recent progress on the genes regulating stomatal density, and discusses the role of stomatal density in plant water use efficiency and the possibility to increase plant water use efficiency, hence bio-water saving by genetically manipulating stomatal density.