• Stomata aperture is key for oxygen and carbon dioxide exchange in plants and essential for photosynthesis.
  • CRAG researchers identified the interplay between two proteins, PIF and KAT1, that are crucial in the regulation of the rhythmic opening/closing of stomata during the day/night cycle.
  • Knowledge of this molecular mechanism is key to understand how plants regulate photosynthesis when challenged by the environment, and it can be useful to prevent crop yield loss under water restrictive and drought conditions.

Plants constantly exchange oxygen (O2) and carbon dioxide (CO2) with the environment, an essential process for photosynthesis. This breathing process occurs thanks to a very specific structure: the stoma. Stomata (from Greek στόμα, “mouth”) are pores found on the surface of leaves that control the gas exchange between the plant and the external atmosphere. Control of stomata aperture is essential to regulate photosynthesis and water use efficiency, and the overall plant physiology. Hence, a precise regulation of stomata dynamics is crucial to understand changes in photosynthesis and yield when plants face environmental challenges, and can have an impact in the agricultural sector.

In this work, published in Nature Communications, the group led by the CSIC researcher at CRAG Elena Monte, has deciphered the molecular mechanism that regulates the rhythmic movements of stomata along the day.

Stomata consist of two paired guard cells (GC) and the pore between them, the size of which depends on the swelling or shrinking of the cells, hence increasing or decreasing the stomatal aperture. Optimal stomatal apertures are determined by the integration of different environmental and internal factors. In general, closing occurs in the dark and under water stress in response to the stress hormone ABA (abscisic acid) to prevent water loss, whereas light induces stomata opening to allow CO2 uptake and O2 release during the day.

To fully understand the mechanisms controlling stomatal movements, CRAG researchers studied the model plant Arabidopsis thaliana in periods of controlled light and dark to monitor stomata aperture and to identify the proteins and genes involved, using molecular biology methods as well as bioinformatics analyses.

Researchers found that a family of proteins named PIFs (Phytochrome Interacting Factors) accumulate at the end of the night period, and that this is a crucial step to induce stomata opening in the morning. PIFs are transcription factors that control the expression of certain genes and, in this case, researchers determined that accumulation of PIFs triggers the expression of KAT1 gene, a guard cell-specific potassium (K+) channel that controls the amount of ions and, hence, the ammount of water entering these cells. In the morning, the presence of light activates KAT1 protein, triggering the intake of potassium ions and the swelling of guard cells, therefore causing stomata to open.

Nil Veciana and Arnau Rovira, co-first authors of the work, highlight the importance of the findings in an experimental set-up “with no water restrictions and, thus, endogenous ABA levels”, conditions in which there is rhythmic stomata aperture and the regulatory response to dark/light cycles can be evaluated.

Actually, this interplay between PIF function and endogenous ABA signaling in the guard cells is key for the dynamic regulation of stomata. During the night, endogenous ABA is necessary to maintain stomata closed, while in the morning, a reduction in ABA is necessary for light-induced and PIF-regulated stomata opening.

Although stomata were known to have rhythmic movements and the proteins responsible for opening-closing were already known, this is the first time that the exact transcriptional mechanism regulating stomata aperture during day/night is elucidated. Moreover, identification of PIF and KAT1 as essential players in this regulation opens up the possibility to new research avenues and possible biotechnology approaches. 

Understanding how the light/dark cycle regulates stomatal aperture could be used to identify targets to optimize plant yield and plant adaptation to different stressors, for example in a water restrictive environment and drought conditions.

Read the paper: Nature Communications

Article source: Center for Research in Agricultural Genomics (CRAG)

Image: Images of leaves of the model plant Arabidopsis thaliana with stomata mainly closed (left) and open (right) (marked with arrow heads). Credit: CRAG