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Plant Physiol, May 2003, Vol. 132, pp. 311-317
Nod Factor and Elicitors Activate Different Phospholipid
Signaling Pathways in Suspension-Cultured Alfalfa Cells1
Martine
den Hartog,
Nathalie
Verhoef, and
Teun
Munnik*
Swammerdam Institute for Life Sciences, Department of Plant
Physiology, University of Amsterdam, Kruislaan 318, NL-1098 SM,
Amsterdam, The Netherlands
Lipo-chitooligosaccharides (Nod factors) are
produced by symbiotic Rhizobium sp. bacteria to elicit
Nod responses on their legume hosts. One of the earliest responses is
the formation of phosphatidic acid (PA), a novel second messenger in
plant cells. Remarkably, pathogens have also been reported to trigger
the formation of PA in nonlegume plants. To investigate how host plants
can distinguish between symbionts and pathogens, the effects of Nod factor and elicitors (chitotetraose and xylanase) on the formation of
PA were investigated in suspension-cultured alfalfa (Medicago sativa) cells. Theoretically, PA can be synthesized via two
signaling pathways, i.e. via phospholipase D (PLD) and via
phospholipase C in combination with diacylglycerol (DAG) kinase.
Therefore, a strategy involving differential radiolabeling with
[32P]orthophosphate was used to determine the
contribution of each pathway to PA formation. In support, PLD activity
was specifically measured by using the ability of the enzyme to
transfer the phosphatidyl group of its substrate to a primary alcohol.
In practice, Nod factor, chitotetraose, and xylanase induced the
formation of PA and its phosphorylated product DAG pyrophosphate within
2 min of treatment. However, whereas phospholipase C and DAG kinase were activated during treatment with all three different compounds, PLD
was only activated by Nod factor. No evidence was obtained for the
activation of phospholipase A2.
1
This work was supported by the Netherlands
Organization for Scientific Research and the Royal Netherlands Academy
of Arts and Sciences.
*
Corresponding author; e-mail munnik{at}science.uva.nl; fax
31-205257934.
© 2003 American Society of Plant Biologists
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