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Plant Physiol, December 1999, Vol. 121, pp. 1291-1298

Changes in Cytosolic pH within Arabidopsis Root Columella Cells Play a Key Role in the Early Signaling Pathway for Root Gravitropism1

Amie Caroline Scott and Nina Strömgren Allen*

Department of Botany, Box 7612, North Carolina State University, Raleigh, North Carolina 27695-7612

Ratiometric wide-field fluorescence microscopy with 1',7'- bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF)-dextran demonstrated that gravistimulation leads to rapid changes in cytoplasmic pH (pHc) in columella cells of Arabidopsis roots. The pHc of unstimulated columella cells in tiers 2 and 3, known sites of graviperception (E.B. Blancaflor, J.B. Fasano, S. Gilroy [1998] Plant Physiol 116: 213-222), was 7.22 ± 0.02 pH units. Following gravistimulation, the magnitude and direction of pHc changes in these cells depended on their location in the columella. Cells in the lower side of tier 2 became more alkaline by 0.4 unit within 55 s of gravistimulation, whereas alkalinization of the cells on the upper side was slower (100 s). In contrast, all cells in tier 3 acidified by 0.4 pH unit within 480 s after gravistimulation. Disrupting these pHc changes in the columella cells using pHc modifiers at concentrations that do not affect root growth altered the gravitropic response. Acidifying agents, including bafilomycin A1, enhanced curvature, whereas alkalinizing agents disrupted gravitropic bending. These results imply that pHc changes in the gravisensing cells and the resultant pH gradients across the root cap are important at an early stage in the signal cascade leading to the gravitropic response.

1 This work was supported by a grant from the National Aeronautics and Space Administration (no. NAGW-4984) and Sigma Xi Grant-in-Aid-of-Research.

* Corresponding author; e-mail nina_allen{at}ncsu.edu; fax 919-515-3436.

© 1999 American Society of Plant Physiologists


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