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The phytohormone abscisic acid (ABA) is a major element involved in apple ( Malus domestica ) production because of its role in seed germination and early seedling development. The WRKY family, which is one of the largest families of transcription factors, plays an important role in ABA signaling in plants. However, the underlying molecular mechanisms of WRKY-mediated ABA sensitivity in apple are poorly understood. A genome-wide transcriptome analysis indicated that  MdWRKY31  is a key factor induced by ABA. Quantitative real-time PCR showed that  MdWRKY31  is induced by ABA in response to PEG4000, which is used to simulate drought. As a transcription factor, MdWRKY31 is localized in the nucleus. Ectopic expression of  MdWRKY31  in  Arabidopsis  and  Nicotiana benthamiana  enhanced plant sensitivity to ABA. Overexpression of  MdWRKY31  in apple roots and apple calli increased sensitivity to ABA, whereas repression of  MdWRKY31  reduced sensitivity to ABA in the roots of ‘Royal Gala’. Electrophoretic mobility shift assays, chromatin immunoprecipitation PCR, and yeast one-hybrid assays indicated that MdWRKY31 directly binds to the promoter of  MdRAV1 . Expression analyses of transgenic apple calli containing  MdWRKY31  and  pMdRAV1::GUS  implied that MdWRKY31 represses the expression of  MdRAV1 . We also found that MdRAV1 binds directly to the promoters of  MdABI3  and  MdABI4  and repressed their expression. Our findings reveal a new important regulatory mechanism of MdWRKY31-MdRAV1-MdABIs in the ABA signaling pathway in apple.

The MdWRKY31 transcription factor binds to the MdRAV1 promoter to mediate ABA sensitivity