The AP2/ERF transcription factor MdDREB2A regulates nitrogen utilisation and sucrose transport under drought stress.

Drought stress is one of the main environmental factors limiting plant growth and development. Plants adapt to changing soil moisture by modifying root architecture, inducing stomatal closure, and inhibiting shoot growth. The AP2/ERF transcription factor DREB2A plays a key role in maintaining plant growth in response to drought stress, but the molecular mechanism underlying this process remains to be elucidated. Here, it was found that overexpression of MdDREB2A positively regulated nitrogen utilisation by interacting with DRE cis-elements of the MdNIR1 promoter. Meanwhile, MdDREB2A could also directly bind to the promoter of MdSWEET12, which may enhance root development and nitrogen assimilation, ultimately promoting plant growth. Overall, this regulatory mechanism provides an idea for plants in coordinating with drought tolerance and nitrogen assimilation to maintain optimal plant growth and development under drought stress.

The apple BTB protein MdBT2 positively regulates MdCOP1 abundance to repress anthocyanin biosynthesis.

The ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1) plays a central role in light-induced anthocyanin biosynthesis. However, the upstream regulatory factors of COP1 remain poorly understood, particularly in horticultural plants. Here, we identified an MdCOP1-interacting protein, BROAD-COMPLEX, TRAMTRACK AND BRIC A BRAC2 (MdBT2), in apple (Malus domestica). MdBT2 is a BTB protein that directly interacts with and stabilizes MdCOP1 by inhibiting self-ubiquitination. Fluorescence observation and cell fractionation assays showed that MdBT2 increased the abundance of MdCOP1 in the nucleus. Moreover, a series of phenotypic analyses indicated that MdBT2 promoted MdCOP1-mediated ubiquitination and degradation of the MdMYB1 transcription factor, inhibiting the expression of anthocyanin biosynthesis genes and anthocyanin accumulation. Overall, our findings reveal a molecular mechanism by which MdBT2 positively regulates MdCOP1, providing insight into MdCOP1-mediated anthocyanin biosynthesis.

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

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.