Methylesterification of cell wall pectin controls the diurnal flower opening times in rice

Flowers are the core reproductive organ of plants,and flowering is essential for cross-pollination.Diurnal flower-opening time is thus a key trait influencing reproductive isolation,hybrid breeding,and thermosta-bility in plants.However,the molecular mechanisms controlling this trait remain unknown.Here,we report that rice Diurnal Flower Opening Time 1(DFOT1)modulates pectin methylesterase(PME)activity to regu-late pectin methylesterification levels of the lodicule cell walls,which affect lodicule swelling to control diurnal flower-opening time.DFOT1 is specifically expressed in the lodicules,and its expression gradually increases with the approach to flowering but decreases with flowering.Importantly,a knockout of DFOT1 showed earlier diurnal flower opening.We demonstrate that DFOT1 interacts directly with multiple PMEs to promote their activity.Knockout of PME40 also resulted in early diurnal flower opening,whereas overex-pression of PME42 delayed diurnal flower opening.Lower PME activity was observed to be associated with higher levels of pectin methylesterification and the softening of cell walls in lodicules,which contribute to the absorption of water by lodicules and cause them to swell,thus promoting early diurnal flower opening.Higher PME activity had the opposite effect.Collectively,our work uncovers a molecular mechanism underlying the regulation of diurnal flower-opening time in rice,which would help reduce the costs of hybrid breeding and improve the heat tolerance of flowering plants by avoiding higher tempera-tures at anthesis.