A rice semi-dwarf variety, IR8, known as “miracle rice” enabled dramatic increases rice production and its widespread adoption averted predicted food shortages in Asia during the 1960s to 1990s. This remarkable achievement was referred to as “green revolution”. The short stature of IR8 was derived from the semi-dwarf gene, sd1, and the sd1 gene contributed significantly to the rice “green revolution”. In this paper, we described the physiological, molecular genetic and biochemical characterization of the sd1 gene. The sd1 mutant contained lower gibberellin (GA) levels than wild-type plants but responded sensitively to exogenous GA. Cloning and sequence analyses revealed that the SD1 gene encoded a GA biosynthetic enzyme, GA20 oxidase. In all of the sd1 mutants tested, nucleotide deletions or substitutions were observed in the GA20 oxidase gene (GA20ox-2), which induced an internal stop codon or single amino acid substitutions, respectively. The sd1 plants, which the wild-type GA20ox-2 gene was introduced showed the normal height. A recombinant GA20ox-2 protein produced from the cDNA clone in E. coli catalyzed the conversion of GA₅₃ to GA₂₀. These results confirmed that SD1 encodes an active GA20 oxidase. The expression of GA20ox-2 was down-regulated by GA in a similar manner to that of some GA20oxs in other plants. The rice genome carried at least two GA 20-oxidase genes (GA20ox-1 and GA20ox-2) and SD1 corresponded to GA20ox-2, which is highly expressed in the leaves and flowers, whereas GA20ox-1 is preferentially expressed in the flowers. The reduced plant height associated with the sd1 alleles was due to the low amount of active GA in leaves, which was caused by a mutation of the GA20ox-2 gene. On the basis of these results, we discussed the importance of GA in the regulation of plant height in crop breeding.