Microtubule dynamics modulate sensing during cold acclimation in grapevine suspension cells

Cold acclimation is of practical relevance, since it can avoid cold-induced damage in various crops. To efficiently activate cold acclimation requires that the chilling stress is perceived and processed efficiently. In the current work, we use a transgenic cell line of V. rupestris expressing a GFP-labelled tubulin to follow the effect of cold acclimation and the relation between microtubules and the expression of the transcription factor Cold Box Factor 4 (CBF4) as molecular readout for adaptive responses to cold stress. We find that chilling induced cold tolerance correlated with increased CBF4 expression. We show that cold acclimation can be achieved through stabilisation of microtubules by taxol, as well as through transient elimination of microtubules by pronamide in the absence of cold stress. Furthermore, results from inhibitor studies indicate that transcriptional activation of CBF4 appears to be under control of calcium influx. We screened a population of the ancestor of V. sylvestris and could identify different clades with strong induction of CBF4, indicative of genetic variation in cold adaptability that can be used for breeding. We summarize our findings into a working model where microtubule dynamics controls the sensitivity of cold induced calcium influx mediating the induction of CBF4 culminating in cold hardening.