Climate change does not impact the water flow of barley at the vegetative stage, ameliorates at anthesis and worsens after subsequent drought episodes.

Climate change will bring the interaction of stresses such as increased temperature and drought under high [CO] conditions. This is likely to impact on crop growth and productivity. This study aimed to (i) determine the response of barley water relations to vegetative and anthesis drought periods under triple interaction conditions, (ii) test the possibility to prime barley plants for drought, and (iii) analyse the involvement of aquaporins in (i) and (ii). The water status of barley was not affected by drought at the vegetative stage, regardless of the environmental conditions. At the anthesis stage, when the water shortage period was more severe, barley plants growing under combined elevated CO and temperature conditions were able to maintain a better water status compared with plants grown under current conditions. Elevated CO and temperature conditions reduced the stomatal conductance and slowed down the plant water flow through a root-leaf hydraulic conductivity coordination. Leaf HvPIP2;1 and HvTIP1;1 aquaporins seemed to play a key role regulating barley's water flow, while leaf and root HvPIP2;5 provided basic level of water flow. At anthesis drought and under future combined conditions, plants showed a reduced cell dehydration and decrease in leaf relative water content compared with plants grown under current conditions. Exposure to a previous drought did not prime the water status of barley plants to a subsequent drought, but instead worsened the response under future conditions. This was due to an imbalance between the roots versus shoot development.