Journal
PHYTOCHEMISTRY
Volume 67, Issue 12, Pages 1232-1242Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.phytochem.2006.04.006
Keywords
Triticum durum; gramineae; durum wheat; copper; lipids; phosphatidic acid; phospholipase D; reactive oxygen species
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Wheat seedlings (Triticum durum Desf.) were incubated in 100 mu M Cu2+ for different periods of time ranging from 1 min up to 16 h. Following metal addition a rapid intake of copper ions into the roots was observed. Cu2+ induced an accumulation of both phosphatidic acid and phosphatidylbutanol within 1 min of incubation, the latter indicating a very rapid induction of phospholipase D (PLD) activity. The highest PLD stimulation was detected after 2 h from copper addition and decreased almost to the initial value at increasing times. Cycloheximide treatment of roots lowered phosphatidylbutanol accumulation because of a reduced PLD activity. The expression profile of a T. durum putative PLD-encoding gene showed a peak after 1 h of treatment as well, indicating that enhanced gene expression contributed to the increase in PLD activity. In the absence of copper ions, roots treated with the G protein activator mastoparan showed increases in phosphatidic acid and phosphatidylbutanol similar to those detected with the metal. PLD activity was also stimulated by cholera toxin. Two putatively G protein alpha subunit encoding sequences were isolated and no significant differences in transcription activity following Cu2+ addition were observed. In copper-treated roots an early production of superoxide generated both by total and membrane-bound NADPH oxidase occurred. The G protein inhibitor suramin as well as the PLD antagonist 1-butanol abolished copper-induced superoxide production. (c) 2006 Elsevier Ltd. All rights reserved.
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