ALA Upregulates MdPTPA Expression to Increase the PP2A Activity and Promote Stomatal Opening in Apple Leaves

Abstract

5-Aminolevulinic acid (ALA) is a new natural plant growth regulator that inhibits abscisic acid (ABA)-induced stomatal closure. Studies have shown that protein phosphatase 2 A (PP2A) is involved in ALA-ABA antagonistically regulating stomatal movement; however, the molecular mechanisms underlying remain unclear. Here, we report that ALA promoted MdPP2A activity and the MdPP2AC expression in the epidermis of apple (Malus × domestica Borkh. cv. Fuji) leaves. Y2H (Yeast two hybrid), BiFC (Bimolecular fluorescence complement), and FLC (Firefly luciferase complementation imaging assay) analysis showed that MdPP2AC interacted with MdPTPA, a phosphortyrosyl phosphatase activator. Furthermore, the transient overexpression or interference-expression of MdPTPA transgenic apple leaves were developed. The results showed that overexpression of MdPTPA promoted stomatal opening by reducing Ca²⁺ and H₂O₂ but increasing flavonols in guard cells. Conversely, when the MdPTPA was silenced in transient transgenic apple leaves, the Ca²⁺, H₂O₂ and flavonols in guard cells and stomatal movement were completely conversed. In the transgenic apple leaves, exogenous ALA stimulated PP2A but repressed SnRK2.6 activity, while the responses are the same as that in the wild type. Therefore, we propose that MdPTPA, which increases the PP2A activity, mediates ALA signaling to promote stomatal opening in apple leaves.

Keywords: ALA; Apple; MdPP2AC; MdPTPA; Stomatal opening.