CBF4 is a unique member of the CBF transcription factor family of Vitis vinifera and Vitis riparia

Abstract

The CBF/DREB1 transcription factors control an important pathway for increased freezing and drought tolerance in plants. We report here the isolation of one CBF/DREB1-like gene, CBF4, from both freezing-tolerant wild grape (Vitis riparia) and freezing-sensitive cultivated grape (Vitis vinifera). The deduced protein in V. riparia is 99% identical to the corresponding protein in V. vinifera; 45-48% to three other Vitis CBF proteins reported earlier and 57% to AtCBF1, and contains CBF-specific amino acid motifs. Agroinfiltration experiments in tobacco leaves revealed that VrCBF4 activates expression from reporter genes driven by a CRT-containing promoter. Expression of the endogenous Vitis CBF4 genes was low at ambient temperature, but enhanced upon exposure to low temperature (4°C). Uncommon for CBF genes, this expression was maintained for several days. No significant difference in expression pattern was observed between V. riparia and V. vinifera. Vitis CBF4 was expressed in both young and mature tissue, in contrast to the previously described Vitis CBF1, 2 and 3. Together, these results suggest that CBF4 represents a second type of CBF in grape that might be more important for the over-wintering of grape plants.

Isolation and expression analysis of CBF4 from <i>Vitis amurensis</i> associated with stress

“…These transcripts remain present for a long period, with lower expression in the leaves than in the other tissues. A similarly cold-induced gene expression was reported for the CBF4 of Vitis riparia and V. vinifera [15]. In agreement with the high identity between their amino acid domains, the expression patterns were almost the same between V. amurensis and V. vinifera under low temperature treatment.…”

“…The N-terminal region of VaCBF4 was found to share the conserved CBF4 domains with that of the other known CBF4 (Figure 1). Vitis CBF4 was proven to be a nuclear protein [15]. Moreover, Va CBF4 was able to map to the CBF4 group in the CBF/DREB subfamily through phylogenetic analysis.…”

“…In agreement with the high identity between their amino acid domains, the expression patterns were almost the same between V. amurensis and V. vinifera under low temperature treatment. Interestingly, under natural conditions, the CBF4 expression of V. vinifera was visible in the leaves [15], whereas that of V. amurensis was not detectable in the leaves, stems, roots, and petioles. This finding was further confirmed by quantitative RT-PCR.…”

“…All experiments were run in triplicates with different synthesizedc DNA fragments. A positive control was provided through a parallel analysis using the grape malate dehydrogenase gene as an internal reference [15], and was calculated relative to a calibrator using the formula 2 −△△Ct .…”

Response, Tolerance and Adaptation to Abiotic Stress of Olive, Grapevine and Chestnut in the Mediterranean Region: Role of Abscisic Acid, Nitric Oxide and MicroRNAs

“…In plants, the Crepeat (CRT)-binding factor/dehydration-responsive element (DRE) binding protein 1 (CBF/DREB1) transcription factors control an important pathway for increased freezing and drought tolerance. Three CBF/DREB1-like genes, CBF 1-3, have been cloned from both freezing-tolerant wild grape (V. riparia) and freezing-sensitive cultivated grape (V. vinifera) (Xiao et al, 2008). The transgenic grapevine over-expressing DREB1b has been proved to have a significantly improved resistance to cold stress (Jin et al, 2009).…”