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bioRxiv · 10.64898/2026.07.10.737675

Stomatal movement in Arabidopsis is driven by guard cell-localized and copper-insensitive CSD1 splice variant

Tsinyk, M.·Hlavackova, K.·Ovecka, M.·Rehak, J.·Sojka, J.·Spundova, M.·Kucerova, Z.·Samaj, J.·Takac, T.·Dvorak, P.
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Abstract

Copper (Cu) is an essential micronutrient whose bioavailability is strongly affected by soil physicochemical properties. During evolution, plants have developed mechanisms to flexibly adjust their metabolism to Cu status. Superoxide dismutases (SODs), including Cu/ZnSOD1 (CSD1) and FeSOD1 (FSD1), are key antioxidant enzymes regulated in Cu dependent manner in Arabidopsis thaliana. Examination of CSD1 cellular distribution and activity revealed that CSD1 is a nuclear and cytosolic SOD whose abundance and activity respond to Cu availability inversely to FSD1. Combined microscopic and biochemical analyses of Cu-dependent dynamics revealed that, unlike FSD1, CSD1 localization in guard cells (GCs) remains independent of Cu availability. CSD1 escapes miR398-mediated regulation in GCs through a cell type-specific splice variant (CSD1.2) that carries an altered miR398-binding site. In silico analyses indicate that this mechanism is also present in crop species. Functionally, the csd1 mutant showed reduced sensitivity to abscisic acid (ABA)-induced stomatal closure, a phenotype rescued by reintroducing CSD1. Biochemical and reactive oxygen species (ROS) level analyses indicate that CSD1.2 most likely acts independently of its canonical enzymatic activity in GCs and functions upstream of the ROS burst in the ABA signaling pathway. Together, we present a novel, cell-type-specific mechanism that safeguards ABA-driven stomatal closure under fluctuating Cu supply.

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Tsinyk, M., Hlavackova, K., Ovecka, M., Rehak, J., Sojka, J., Spundova, M., Kucerova, Z., Samaj, J., Takac, T., Dvorak, P.. 2026-07-10. Stomatal movement in Arabidopsis is driven by guard cell-localized and copper-insensitive CSD1 splice variant. https://doi.org/10.64898/2026.07.10.737675

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