The Hsp20 genes represent the most abundant small heat shock proteins (sHSPs) in plants. Hsp20 gene family has been shown to be involved in preventing heat shock and promoting resistance to environmental stress factors, but very little is known about this gene family in rice. Here, we report the identification and characterization of 39 OsHsp20 genes in rice, describing the gene structure, gene expression, genome localization, and phylogenetic relationship of each member. We have used RT-PCR to perform a characterization of the normal and heat shock-induced expression of selective OsHsp20 genes. A genome-wide microarray based gene expression analysis involving 25 stages of vegetative and reproductive development in three rice cultivars has revealed that 36 OsHsp20 genes were expressed in at least one of the experimental stages studied. Among these, transcripts of OsHsp20 were accumulated differentially during vegetative and reproductive developmental stages and preferentially down-regulated in Shanyou 63. In addition, OsHsp20 genes were identified as showing prominent heterosis in family-level expression. Our results suggest that the expression patterns of the OsHsp20 genes are diversified not only in developmental stages but also in variety level.