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Background  In winter barley plants, vernalization and photoperiod cues have to be integrated to promote flowering. Plant development and expression of different flowering promoter ( HvVRN1 ,  HvCO2 ,  PPD-H1 ,  HvFT1 ,  HvFT3 ) and repressor ( HvVRN2 ,  HvCO9  and  HvOS2 ) genes were evaluated in two winter barley varieties under: (1) natural increasing photoperiod, without vernalization, and (2) under short day conditions in three insufficient vernalization treatments. These challenging conditions were chosen to capture non-optimal and natural responses, representative of those experienced in the Mediterranean area.    Results  In absence of vernalization and under increasing photoperiods,  HvVRN2  expression increased with day-length, mainly between 12 and 13 h photoperiods in our latitudes. The flowering promoter gene in short days,  HvFT3 , was only expressed after receiving induction of cold or plant age, which was associated with low transcript levels of  HvVRN2  and  HvOS2 . Under the sub-optimal conditions here described, great differences in development were found between the two winter barley varieties used in the study. Delayed development in ‘Barberousse’ was associated with increased expression levels of  HvOS2 . Novel variation for  HvCO9  and  HvOS2  is reported and might explain such differences.    Conclusions  The balance between the expression of flowering promoters and repressor genes regulates the promotion towards flowering or the maintenance of the vegetative state.  HvOS2 , an ortholog of  FLC , appears as a strong candidate to mediate in the vernalization response of barley. Natural variation found would help to exploit the plasticity in development to obtain better-adapted varieties for current and future climate conditions.    Electronic supplementary material  The online version of this article (10.1186/s12870-019-1727-9) contains supplementary material, which is available to authorized users.

Fine-tuning of the flowering time control in winter barley: the importance of HvOS2 and HvVRN2 in non-inductive conditions