The apple gene responsible for columnar tree shape reduces the abundance of biologically active gibberellin

Ectopic expression of the apple 2-oxoglutarate-dependent dioxygenase (DOX, 2ODD) gene, designated MdDOX-Co, is thought to cause the columnar shape of apple trees. However, the mechanism underlying the formation of such a unique tree shape remains unclear. To solve this problem, we demonstrated that Arabidopsis thaliana overexpressing MdDOX-Co contained reduced levels of biologically active gibberellin (GA) compared with wild type. In summary: (i) with biochemical approaches, the gene product MdDOX-Co was shown to metabolize active GA A₄ (GA₄ ) to GA₅₈ (12-OH-GA₄ ) in vitro. MdDOX-Co also metabolized its precursors GA₁₂ and GA₉ to GA₁₁₁ (12-OH-GA₁₂ ) and GA₇₀ (12-OH-GA₉ ), respectively; (ii) Of the three 12-OH-GAs, GA₅₈ was still active physiologically, but not GA₇₀ or GA₁₁₁ ; (iii) Arabidopsis MdDOX-Co OE transformants converted exogenously applied deuterium-labeled (d₂ )-GA₁₂ to d₂ -GA₁₁₁ but not to d₂ -GA₅₈ , whereas transformants converted applied d₂ -GA₉ to d₂ -GA₅₈ ; (iv) GA₁₁₁ is converted poorly to GA₇₀ by GA 20-oxidases in vitro when GA₁₂ is efficiently metabolized to GA₉ ; (v) no GA₅₈ was detected endogenously in MdDOX-Co OE transformants. Overall, we conclude that 12-hydroxylation of GA₁₂ by MdDOX-Co prevents the biosynthesis of biologically active GAs in planta, resulting in columnar phenotypes.