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First published online June 6, 2008; 10.1104/pp.108.119529 Plant Physiology 147:1659-1674 (2008) © 2008 American Society of Plant Biologists OPEN ACCESS ARTICLE
A Class I ADP-Ribosylation Factor GTPase-Activating Protein Is Critical for Maintaining Directional Root Hair Growth in Arabidopsis1,[W],[OA]Plant Biology Division, Samuel Roberts Noble Foundation, Ardmore, Oklahoma 73401
Membrane trafficking and cytoskeletal dynamics are important cellular processes that drive tip growth in root hairs. These processes interact with a multitude of signaling pathways that allow for the efficient transfer of information to specify the direction in which tip growth occurs. Here, we show that AGD1, a class I ADP ribosylation factor GTPase-activating protein, is important for maintaining straight growth in Arabidopsis (Arabidopsis thaliana) root hairs, since mutations in the AGD1 gene resulted in wavy root hair growth. Live cell imaging of growing agd1 root hairs revealed bundles of endoplasmic microtubules and actin filaments extending into the extreme tip. The wavy phenotype and pattern of cytoskeletal distribution in root hairs of agd1 partially resembled that of mutants in an armadillo repeat-containing kinesin (ARK1). Root hairs of double agd1 ark1 mutants were more severely deformed compared with single mutants. Organelle trafficking as revealed by a fluorescent Golgi marker was slightly inhibited, and Golgi stacks frequently protruded into the extreme root hair apex of agd1 mutants. Transient expression of green fluorescent protein-AGD1 in tobacco (Nicotiana tabacum) epidermal cells labeled punctate bodies that partially colocalized with the endocytic marker FM4-64, while ARK1-yellow fluorescent protein associated with microtubules. Brefeldin A rescued the phenotype of agd1, indicating that the altered activity of an AGD1-dependent ADP ribosylation factor contributes to the defective growth, organelle trafficking, and cytoskeletal organization of agd1 root hairs. We propose that AGD1, a regulator of membrane trafficking, and ARK1, a microtubule motor, are components of converging signaling pathways that affect cytoskeletal organization to specify growth orientation in Arabidopsis root hairs.
1 This work was supported by the Samuel Roberts Noble Foundation and by the Oklahoma Center for the Advancement of Science and Technology (grant no. PSB08–003 to E.B.B.). The confocal microscopes used in this study were acquired through Multi-User Equipment and Major Research Instrumentation grants from the National Science Foundation (grant nos. DBI–0400580 and DBI–0722635 to E.B.B.). The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Elison B. Blancaflor (eblancaflor{at}noble.org). [W] The online version of this article contains Web-only data. [OA] Open Access articles can be viewed online without a subscription. www.plantphysiol.org/cgi/doi/10.1104/pp.108.119529 * Corresponding author; e-mail eblancaflor{at}noble.org. Received March 23, 2008; accepted May 27, 2008; published June 6, 2008. This article has been cited by other articles:
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