Overexpression of the ArabidopsisCBF3Transcriptional Activator Mimics Multiple Biochemical Changes Associated with Cold Acclimation

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Abstract
We further investigated the role of the ArabidopsisCBF regulatory genes in cold acclimation, the process whereby certain plants increase in freezing tolerance upon exposure to low temperature. The CBF genes, which are rapidly induced in response to low temperature, encode transcriptional activators that control the expression of genes containing the C-repeat/dehydration responsive element DNA regulatory element in their promoters. Constitutive expression of either CBF1 orCBF3 (also known as DREB1b andDREB1a, respectively) in transgenic Arabidopsis plants has been shown to induce the expression of target COR(cold-regulated) genes and to enhance freezing tolerance in nonacclimated plants. Here we demonstrate that overexpression ofCBF3 in Arabidopsis also increases the freezing tolerance of cold-acclimated plants. Moreover, we show that it results in multiple biochemical changes associated with cold acclimation:CBF3-expressing plants had elevated levels of proline (Pro) and total soluble sugars, including sucrose, raffinose, glucose, and fructose. Plants overexpressing CBF3 also had elevated P5CS transcript levels suggesting that the increase in Pro levels resulted, at least in part, from increased expression of the key Pro biosynthetic enzyme Δ1-pyrroline-5-carboxylate synthase. These results lead us to propose that CBF3 integrates the activation of multiple components of the cold acclimation response.