Transcription factor VRT2 reinitiates vernalization when interrupted by warm temperatures in a temperate grass model

Vernalisation-responsive plants use cold as a cue to monitor the passing of winter. Winter cereals can remember how much cold they have experienced, even when winter is punctuated by warm days. However, in a seemingly unnatural process called 'devernalisation', hot temperatures can erase winter memory. Previous studies in bread wheat (Triticum aestivum) have implicated the MADS-box transcription factor VEGETATIVE TO REPRODUCTIVE TRANSITION 2 (VRT2) in vernalisation based on transcriptional behaviour and ectopic expression. Here, we characterised three BdVRT2 loss-of-function alleles in the temperate model grass Brachypodium distachyon. In addition to extended vernalisation requirements, mutants showed delayed flowering relative to wild-type plants when exposed only briefly to warm temperatures after partial vernalisation, with flowering being unaffected when vernalisation was saturating. Together, these data suggest a role for BdVRT2 in both vernalisation and in its re-initiation when interrupted by warm temperatures. In controlled constant conditions, BdVRT2 transcription was not strongly affected by vernalisation or devernalisation. Yet, by monitoring BdVRT2 expression in seasonally varying and fluctuating conditions in an unheated greenhouse, we observed strong upregulation, suggesting that its transcription is regulated by fluctuating vernalising-devernalising conditions. Our data suggest that devernalisation by hot temperatures is not a peculiarity of domesticated cereal crops but is the extreme of the reversibility of vernalisation by warm temperatures and has broader biological relevance across temperate grasses.