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Plant and Cell Physiology, 2003, Vol. 44, No. 7 687-696
© 2003 Oxford University Press

The Effects of the Phospholipase D-Antagonist 1-Butanol on Seedling Development and Microtubule Organisation in Arabidopsis

John Gardiner1,4,5, David A. Collings2,5, John D. I. Harper1,3 and Jan Marc1,6

1 School of Biological Sciences, The University of Sydney, Sydney NSW 2006, Australia
2 Research School of Biological Sciences, The Australian National University, Canberra ACT 2601, Australia
3 Farrer Centre, School of Agriculture, Charles Sturt University, Wagga Wagga, NSW 2678, Australia

The organisation of plant microtubules into distinct arrays during the cell cycle requires interactions with partner proteins. Having recently identified a 90-kDa phospholipase D (PLD) that associates with microtubules and the plasma membrane [Gardiner et al. (2001) Plant Cell 13: 2143], we exposed seeds and young seedlings of Arabidopsis to 1-butanol, a specific inhibitor of PLD-dependent production of the signalling molecule phosphatidic acid (PA). When added to agar growth media, 0.2% 1-butanol strongly inhibited the emergence of the radicle and cotyledons, while 0.4% 1-butanol effectively blocked germination. When normal seedlings were transferred onto media containing 0.2% and 0.4% 1-butanol, the inhibitor retarded root growth by about 40% and 90%, respectively, by reducing cell elongation. Inhibited plants showed significant swelling in the root elongation zone, bulbous or branched root hairs, and modified cotyledon morphology. Confocal immunofluorescence microscopy of root tips revealed that 1-butanol disrupted the organisation of interphase cortical microtubules. Butanol isomers that do not inhibit PLD-dependent PA production, 2- and 3-butanol, had no effect on seed germination, seedling growth, or microtubule organisation. We propose that production of PA by PLD may be required for normal microtubule organisation and hence normal growth in Arabidopsis.

4 Current address: School of Biological Sciences, The University of Manchester, Manchester M13 9PT, U.K.

5 John Gardiner and David Collings contributed equally to this work.

6 Corresponding author: E-mail, jmarc{at}bio.usyd.edu.au; Fax, +61-2-9351-4771.


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