CBF gene expression in peach leaf and bark tissues is gated by a circadian clock

Abstract

CBF (C-repeat Binding Factor) transcription factors are part of the AP2/ERF (Apetala2-ethylene responsive factor) domain family of DNA-binding proteins that recognize a C-repeat response cis-acting element that regulates a number of cold-responsive genes (CBF regulon). Induction of CBF gene expression by low temperature in Arabidopsis has been shown to be gated by a circadian clock. In peach (Prunus persica L.), five CBF genes are arranged in tandem on scaffold (linkage group) 5 of the peach genome. Since CBF gene regulation has been shown to be more complex in woody plants than herbaceous plants, the present study was conducted to determine if temperature-modulated CBF gene expression in peach leaf and bark tissues was also influenced by a circadian clock. One-year-old 'Loring' peach trees grafted on 'Bailey' rootstocks were entrained to a 12-h day/12-h night photoperiod at 25 °C. After 2 weeks, trees were exposed to 4 °C under continuous light for up to 48 h beginning at either subjective dawn + 4 h (ZT4; where ZT is Zeitgeber time) or subjective dawn + 16 h (ZT16) with leaf and bark tissues harvested at various time points. Gene expression of the five peach CBF genes and a DREB2 gene was assessed by real-time quantitative polymerase chain reaction. Results revealed a distinct gating of CBF gene expression by a circadian clock for four CBF genes in both leaf and bark tissues. CBF genes were highly induced by 4 °C in ZT4 leaf samples with expression peaking at 6-24 h depending on the specific CBF gene. In contrast, CBF gene expression was highly attenuated in leaf, and to a lesser extent in bark, samples exposed to 4 °C at ZT16. These results are similar to reports for Arabidopsis. Further experiments were conducted to verify environmental influence on the induction of CBF and DREB2 genes. In contrast to DREB2 genes from other dicots, the peach DREB2 ortholog was induced by both low temperature and dehydration. Induction of the peach CBFs and DREB2 by either low temperature or dehydration corresponded with regulatory motifs present in their promoter sequences. Low temperature and dehydration induction data for three peach dehydrin genes indicated that the regulation of these genes in peach is complex, with individual dehydrin gene expression being correlated with the expression of one or more CBF genes.