Gibberellins Inhibit Flavonoid Biosynthesis and Promote Nitrogen Metabolism in Medicago truncatula

Bioactive gibberellic acids (GAs) are diterpenoid plant hormones that are biosynthesized through complex pathways and control various aspects of growth and development. Although GA biosynthesis has been intensively studied, the downstream metabolic pathways regulated by GAs have remained largely unexplored. We investigated Tnt1 retrotransposon insertion mutant lines of Medicago truncatula with a dwarf phenotype by forward and reverse genetics screening and phylogenetic, molecular, biochemical, proteomic and metabolomic analyses. Three Tnt1 retrotransposon insertion mutant lines of the gibberellin 3-beta-dioxygenase 1 gene (GA3ox1) with a dwarf phenotype were identified, in which the synthesis of GAs (GA(3) and GA(4)) was inhibited. Phenotypic analysis revealed that plant height, root and petiole length of ga3ox1 mutants were shorter than those of the wild type (Medicago truncatula ecotype R108). Leaf size was also much smaller in ga3ox1 mutants than that in wild-type R108, which is probably due to cell-size diminution instead of a decrease in cell number. Proteomic and metabolomic analyses of ga3ox1/R108 leaves revealed that in the ga3ox1 mutant, flavonoid isoflavonoid biosynthesis was significantly up-regulated, while nitrogen metabolism was down-regulated. Additionally, we further demonstrated that flavonoid and isoflavonoid biosynthesis was induced by prohexadione calcium, an inhibitor of GA3ox enzyme, and inhibited by exogenous GA(3). In contrast, nitrogen metabolism was promoted by exogenous GA(3) but inhibited by prohexadione calcium. The results of this study further demonstrated that GAs play critical roles in positively regulating nitrogen metabolism and transport and negatively regulating flavonoid biosynthesis through GA-mediated signaling pathways in leaves.

Automated summary

The study identified mutant lines with a dwarf phenotype due to Tnt1 retrotransposon insertion in the gibberellin 3-beta-dioxygenase 1 gene (GA3ox1), inhibiting the synthesis of bioactive gibberellic acids (GAs). Analysis revealed shorter plant height, root, and petiole length in mutants compared to the wild type, with smaller leaf size attributed to cell-size diminution. Proteomic and metabolomic analyses indicated up-regulation of flavonoid isoflavonoid biosynthesis and down-regulation of nitrogen metabolism in the mutants, suggesting a critical role of GAs in regulating nitrogen metabolism positively and flavonoid biosynthesis negatively in leaves.