Molecular characterization of three GIBBERELLIN-INSENSITIVE DWARF1 homologous genes in hexaploid wheat

GIBBERELLIN-INSENSITIVE DWARF1 (GID1) functions as a gibberellin (GA) receptor and is a key component in the GA signaling pathway. In this paper, three TaGID1 genes, orthologous to rice OsGID1 (the first identified GA receptor GID1 gene), were isolated from hexaploid wheat using homology cloning. Like OsGID1, the three homologous TaGID1 genes consisted of two exons and one intron. Physical location analyses using nullisomic-tetrasomic and deletion lines derived from the wheat cultivar Chinese Spring revealed that the three homologous TaGID1 genes reside in the chromosome bins 1AL3-0.61-1, 1BL1-0.47-0.69, and 1DL2-0.41-1. Accordingly, they were named TaGID1-A1, TaGID1-B1, and TaGID1-D1, respectively. The expression patterns of the three TaGID1 genes were determined by real-time PCR in various wheat tissues at the heading stage, including flag leaves, young spikes, peduncles, and the third and fourth internodes. The three TaGID1 genes had similar transcript patterns, and all exhibited greater expression in flag leaves than in the other tissues. Moreover, they were all down-regulated after treatment with exogenous gibberellic acid (GA(3)) in young seedlings, suggesting a feedback regulation of TaGID1 in wheat. Yeast two-hybrid assays demonstrated strong interactions between each putative TaGID1 and the wheat DELLA proteins RHT-A1, RHT-B1, and RHT-D1 in the presence of GA(3), and weak interactions in the absence of GA(3) in yeast cells. Furthermore, over-expression of each TaGID1 gene in the Arabidopsis double mutant gid1a/1c partially rescued the dwarf phenotype. These results suggest that the three TaGID1 homologous genes are all functional GA receptor genes in wheat.