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>The Arabidopsis KINβγ Subunit of the SnRK1 Complex Regulates Pollen Hydration on the Stigma by Mediating the Level of Reactive Oxygen Species in Pollen
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The Arabidopsis KINβγ Subunit of the SnRK1 Complex Regulates Pollen Hydration on the Stigma by Mediating the Level of Reactive Oxygen Species in Pollen
Pollen–stigma interactions are essential for pollen germination. The highly regulated process of pollen germination includes pollen adhesion, hydration, and germination on the stigma. However, the internal signaling of pollen that regulates pollen–stigma interactions is poorly understood. KINβγ is a plant-specific subunit of the SNF1-related protein kinase 1 complex which plays important roles in the regulation of plant development. Here, we showed that KINβγ was a cytoplasm- and nucleus-localized protein in the vegetative cells of pollen grains in Arabidopsis . The pollen of the Arabidopsis kinβγ mutant could not germinate on stigma, although it germinated normally in vitro . Further analysis revealed the hydration of kinβγ mutant pollen on the stigma was compromised. However, adding water to the stigma promoted the germination of the mutant pollen in vivo , suggesting that the compromised hydration of the mutant pollen led to its defective germination. In kinβγ mutant pollen, the structure of the mitochondria and peroxisomes was destroyed, and their numbers were significantly reduced compared with those in the wild type. Furthermore, we found that the kinβγ mutant exhibited reduced levels of reactive oxygen species (ROS) in pollen. The addition of H_(2)O_(2) in vitro partially compensated for the reduced water absorption of the mutant pollen, and reducing ROS levels in pollen by overexpressing Arabidopsis CATALASE 3 resulted in compromised hydration of pollen on the stigma. These results indicate that Arabidopsis KINβγ is critical for the regulation of ROS levels by mediating the biogenesis of mitochondria and peroxisomes in pollen, which is required for pollen–stigma interactions during pollination. Author Summary After landing on the stigma, pollen grains germinate and create pollen tubes following adhesion and hydration processes, during which pollen–stigma interactions determine whether the pollen grains can germinate on the stigma. In recent years, the interaction mechanisms between the pollen and stigma have been studied extensively at the cellular and molecular level in self-incompatibility systems. However, few studies have focused on pollen–stigma interactions during self-compatible pollination. Arabidopsis thaliana provides an excellent system to study the interaction mechanisms between the pollen and stigma during self-compatible pollination. KINβγ is a plant-specific subunit of the SNF1-related protein kinase 1 complex. In this study, we identified an Arabidopsis kinβγ mutant showing defective pollen germination on the surface of the stigma but not on the culture medium, which resulted from the compromised hydration of pollen on the stigma. Further analysis revealed that the biogenesis of mitochondria and peroxisomes was impaired in this mutant, which reduced the levels of reactive oxygen species (ROS) in pollen. Application of H_(2)O_(2)recovered the capability of pollen to undergo hydration in vitro . These results suggest that ROS signaling is involved in the regulation of pollen–stigma interactions during pollination. This study provides new insights into the mechanism underlying pollen–stigma interactions in self-compatible plant species.
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