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Keywords:

  • 14-3-3;
  • barley;
  • ABA responsive element binding factor (ABF/AREB);
  • ABA;
  • seed germination;
  • yeast two-hybrid

Summary

Proteins of the 14-3-3 family have well-defined functions as regulators of plant primary metabolism and ion homeostasis. However, neither their function nor action mechanism in plant hormonal signaling have been fully addressed. Here we show that abscisic acid (ABA) affects both expression and protein levels of five 14-3-3 isoforms in embryonic barley roots. As ABA prolongs the presence of 14-3-3 proteins in the elongating radicle, we tested whether 14-3-3s are instrumental in ABA action using RNA interference. Transient co-expression of 14-3-3 RNAi constructs along with an ABA-responsive promoter showed that each 14-3-3 is functional in generating an ABA response. In a yeast two-hybrid screen, we identified three new 14-3-3 interactors that belong to the ABF protein family. Moreover, using a yeast two-hybrid assay, we show that the transcription factor HvABI5, which binds to cis-acting elements of the ABA-inducible HVA1 promoter, interacts with three of the five 14-3-3s. Our analyses identify two 14-3-3 binding motifs in HvABI5 that are essential for 14-3-3 binding and proper in vivo trans-activation activity of HvABI5. In line with these results, 14-3-3 silencing effectively blocks trans-activation. Our results indicate that 14-3-3 genes/proteins are not only under the control of ABA, but that they control ABA action as well.