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First published online January 26, 2007; 10.1105/tpc.106.041426

The Plant Cell 19:281-295 (2007)
© 2007 American Society of Plant Biologists

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Arabidopsis PLD{zeta}2 Regulates Vesicle Trafficking and Is Required for Auxin Response[W]

Gang Li and Hong-Wei Xue1

National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, and Partner Group of the Max-Planck-Institute of Molecular Plant Physiology on Plant Molecular Physiology and Signal Transduction, 200032 Shanghai, People's Republic of China

1 To whom correspondence should be addressed. E-mail hwxue{at}sibs.ac.cn; fax 86-21-54924060.

Phospholipase D (PLD) and its product, phosphatidic acid (PA), play key roles in cellular processes, including stress and hormonal responses, vesicle trafficking, and cytoskeletal rearrangements. We isolated and functionally characterized Arabidopsis thaliana PLD{zeta}2, which is expressed in various tissues and enhanced by auxin. A PLD{zeta}2-defective mutant, pld{zeta}2, and transgenic plants deficient in PLD{zeta}2 were less sensitive to auxin, had reduced root gravitropism, and suppressed auxin-dependent hypocotyl elongation at 29°C, whereas transgenic seedlings overexpressing PLD{zeta}2 showed opposite phenotypes, suggesting that PLD{zeta}2 positively mediates auxin responses. Studies on the expression of auxin-responsive genes and observation of the ß-glucuronidase (GUS) expression in crosses between pld{zeta}2 and lines containing DR5-GUS indicated that PLD{zeta}2, or PA, stimulated auxin responses. Observations of the membrane-selective dye FM4-64 showed suppressed vesicle trafficking under PLD{zeta}2 deficiency or by treatment with 1-butanol, a PLD-specific inhibitor. By contrast, vesicle trafficking was enhanced by PA or PLD{zeta}2 overexpression. Analyses of crosses between pld{zeta}2 and lines containing PIN-FORMED2 (PIN2)–enhanced green fluorescent protein showed that PLD{zeta}2 deficiency had no effect on the localization of PIN2 but blocked the inhibition of brefeldin A on PIN2 cycling. These results suggest that PLD{zeta}2 and PA are required for the normal cycling of PIN2-containing vesicles as well as for function in auxin transport and distribution, and hence auxin responses.




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