In this study, five genes encoding NACA were identified in rice, and physico-chemical properties, structure, localization and expression pattern were analyzed. Subsequently, the subcellular localization of NACA2 and its biological function in the response to drought were studied. The results showed that: (1) the five NACA proteins contained many conserved motifs. Both NAC domain and UBA domain were found in NACA1-NACA4, but not in NACA5. The phylogenetic relationship of amino acid sequence in different plants was highly consistent with the evolution of species. (2) Analysis of expression pattern showed that the expression level of NACA1, NACA2 and NACA3 was very high in different tissues, especially in reproductive organs. The transcripts of NACA4 and NACA5 had a low level in all tissues of rice. The expression of NACA was induced by mannitol, abscisic acid (ABA), jasmonic acid (JA), NaCl, salicylic acid (SA) and cold stress, and the expression changes of NACA2 and NACA5 were the most obvious. (3) The analysis of subcellular localization showed that NACA2 located in nucleus and cytoplasm. (4) Overexpression of NACA2 slowed down the water loss rate of leaves and enhanced the resistance of transgenic Arabidopsis to drought and osmotic stresses. Our study suggested that NACA2 positively regulated the tolerance of plant to drought, which will lay a foundation for further study on the biological function of NACA gene in rice.