gid1, a gibberellin‐insensitive dwarf mutant, shows altered regulation of probenazole‐inducible protein (PBZ1) in response to cold stress and pathogen attack

ABSTRACT A recessive gibberellin (GA)‐insensitive dwarf mutant of rice, gibberellin‐insensitive dwarf1 ( gid1 ), has been identified, which shows a severe dwarf phenotype and contains high concentrations of endogenous GA. To elucidate the function of gid1 , proteins regulated downstream of gid1 were analysed using a proteomic approach. Proteins extracted from suspension‐cultured cells of gid1 and its wild type were separated by two‐dimensional polyacrylamide gel electrophoresis (2D‐PAGE). Of a total of 962 proteins identified from the suspension‐cultured cells, 16 were increased and 14 were decreased in gid1 compared with its wild type. Among the proteins hyper‐accumulated in gid1 were osmotin, triosephosphate isomerase, probenazole inducible protein (PBZ1) and pathogenesis‐related protein 10. Of these four genes, only the expression of PBZ1 was increased by exogenous GA 3 application. Expression of this gene was also enhanced in shoots of the wild type by cold stress or by rice blast fungus infection. Under normal growth conditions, there was more PBZ1 protein in gid1 than in the wild type. In addition, gid1 showed increased tolerance to cold stress and resistance to blast fungus infection. The ent ‐copalyl diphosphate synthase ( OsCPS ) genes, which encode enzymes at the branch point between GA and phytoalexin biosynthesis, were expressed differentially in gid1 relative to the wild type. Specifically, OsCPS1 , which encodes an enzyme in the GA biosynthesis pathway, was down‐regulated and OsCPS2 and OsCPS4 , which encode enzymes in phytoalexin biosynthesis, were up‐regulated in gid1 . These results suggest that the expression of PBZ1 is regulated by GA signalling and stress stimuli, and that gid1 is involved in tolerance to cold stress and resistance to blast fungus.