VOZ-dependent priming of salicylic acid-dependent defense against Rhizopus stolonifer by β-aminobutyric acid requires the TCP protein TCP2 in peach fruit

Abstract

Vascular plant one-zinc finger (VOZ) transcription factors (TFs) play crucial roles in plant immunity. Nevertheless, how VOZs modulate defense signaling in response to elicitor-induced resistance is not fully understood. Here, the defense elicitor β-aminobutyric acid (BABA) resulted in the visible suppression of Rhizopus rot disease of peach fruit caused by Rhizopus stolonifer. Defense priming by BABA was notably associated with increased levels of salicylic acid (SA) and SA-dependent gene expression. Data-independent acquisition proteomic analysis revealed that two VOZ proteins (PpVOZ1 and PpVOZ2) were substantially upregulated in BABA-induced resistance (BABA-IR). Furthermore, the interaction of PpVOZ1 and PpVOZ2 and their potential target of the TEOSINTE-BRANCHED1/CYCLOIDEA/PCF (TCP)-family protein PpTCP2 screened from protein–protein interaction networks was confirmed by yeast two-hybrid (Y2H), luciferase complementation imaging and glutathione S-transferase pull-down assays. Furthermore, subcellular localization, yeast one-hybrid, electrophoretic mobility shift assay and dual-luciferase reporter assays demonstrated that nuclear localization of both PpVOZ1 and PpVOZ2 was critical for their contribution to BABA-IR, as these proteins potentiated the PpTCP2-mediated transcriptional activation of isochorismate synthase genes (ICS1/2). The overexpression of both PpVOZ1 and PpVOZ2 could activate the transcription of SA-dependent genes and provide disease resistance in transgenic Arabidopsis. In contrast, the ppvoz1^cas9 and ppvoz2^cas9 loss-of-function mutations and the voz1^cas9 voz2^cas9 double mutation attenuated BABA-IR against R. stolonifer. Therefore, the three identified positive TFs, PpVOZ1, PpVOZ2, and PpTCP2, synergistically contribute to the BABA-activated priming of systemic acquired resistance in postharvest peach fruit by a VOZ-TCP-ICS regulatory module.