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bioRxiv · 10.1101/2021.01.05.425399

The AFB1 auxin receptor controls rapid auxin signaling and root growth through membrane depolarization in Arabidopsis thaliana

Serre, N. B.·Kralik, D.·Yun, P.·Shabala, S.·Slouka, Z.·Fendrych, M.
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Abstract

The existence of an electric gradient across membranes is essential for a cell operation. In plants, application of the growth regulator auxin (IAA) causes almost instantaneous membrane depolarization in various cell types, making membrane depolarization a hallmark of the rapid non-transcriptional responses to IAA. Auxin triggers rapid root growth inhibition; a process that underlies gravitropic bending. The growth and depolarization responses to auxin show remarkable similarities in dynamics, requirement of auxin influx and the involvement of the TIR1/AFB auxin coreceptors, but whether auxin-induced depolarization participates in root growth inhibition remains unanswered. Here, we established a toolbox to dynamically visualize membrane potential in vivo in Arabidopsis thaliana roots by combining the DISBAC2(3) fluorescent probe with microfluidics and vertical stage microscopy. This way we show that auxin-induced membrane depolarization tightly correlates with rapid root growth inhibition and that the cells of the transition zone/early elongation zone are the most responsive to auxin. Further, we demonstrate that auxin cycling in and out of the cells through AUX1 influx and PIN2 efflux is not essential for membrane depolarization and rapid root growth inhibition but acts as a facilitator of these responses. The rapid membrane depolarization by auxin instead strictly depends on the AFB1 auxin receptor, while the other TIR1/AFB paralogues contribute to this response. The lack of membrane depolarization in the afb1 mutant explains the lack of the immediate root growth inhibition. Finally, we show that AFB1 is required for the rapid depolarization and rapid growth inhibition of cells at the lower side of the gravistimulated root. These results are instrumental in understanding the physiological significance of membrane depolarization for the gravitropic response of the root and clarify the role of AFB1 as the receptor central for rapid auxin responses, adding another piece to the puzzle in understanding the biology of the phytohormone auxin.

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Serre, N. B., Kralik, D., Yun, P., Shabala, S., Slouka, Z., Fendrych, M.. 2021-01-06. The AFB1 auxin receptor controls rapid auxin signaling and root growth through membrane depolarization in Arabidopsis thaliana. https://doi.org/10.1101/2021.01.05.425399

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