The transcription factor VaMYC2 from Chinese wild Vitis amurensis enhances cold tolerance of grape (V. vinifera) by up-regulating VaCBF1 and VaP5CS

Abstract

Cultivated grapes, one of the most important fruit crops in the world, are sensitive to low temperature. Since Chinese wild grape Vitis amurensis is highly tolerant to cold, it is imperative to study and utilize its cold-tolerance genes for molecular breeding. Here, a VaMYC2 gene from V. amurensis was cloned, and its function was investigated by expressing VaMYC2 in the cold-sensitive V. vinifera cultivar ‘Thompson Seedless’. The expression of VaMYC2 could be induced by cold stress, methyl jasmonate and ethylene treatment, but was inhibited by abscisic acid in leaves of V. amurensis. When transgenic grape lines expressing VaMYC2 were subjected to cold stress (−1 °C) for 41 h, the transgenic lines showed less freezing injury and lower electrolyte leakage and malondialdehyde content, but higher contents of soluble sugars, soluble proteins and proline, and antioxidant enzyme activities compared with wild-type. Moreover, the expression of some cold-tolerance related genes increased in transgenic lines. Besides, the interactions of VaMYC2 with VaJAZ1 and VaJAZ7B were confirmed by yeast two-hybrid and bimolecular fluorescence complementation assays. Yeast one-hybrid and dual luciferase assays showed that VaMYC2 can bind to the promoters of VaCBF1 and VaP5CS and activate their expressions. In conclusion, expression of VaMYC2 in V. vinifera enhances cold tolerance of transgenic grapes which is attributed to enhanced accumulation of osmotic regulatory substances, cell membrane stability, antioxidant enzyme activity, and expression of cold tolerance-related genes. Also, VaMYC2 interacts with VaJAZ1 and VaJAZ7, and activates the expression of VaCBF1 and VaP5CS to mediate cold tolerance in grapes.