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Plant and Cell Physiology Advance Access published online on February 2, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci065
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received December 20, 2004
Accepted January 14, 2005

Regular Paper

Elicitor-Induced Activation of Phospholipases Plays An Important Role for the Induction of Defense Responses in Suspension-Cultured Rice Cells

Takeshi Yamaguchi 1, Eiichi Minami 2, Jun Ueki 3, and Naoto Shibuya 4*

1 Department of Rice Research, National Agricultural Research Center, National Agricultural Bio-oriented Research Organization, Joetsu, Niigata 943-0193, Japan
2 Biochemistry Department, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan
3 Plant Innovation Center, Japan Tobacco Inc., 700 Higashibara, Iwata, Shizuoka 438-0802, Japan
4 Department of Life Sciences, Faculty of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571, Japan

* To whom correspondence should be addressed.
Naoto Shibuya, E-mail: shibuya{at}isc.meiji.ac.jp


  Abstract

Biphasic generation of reactive oxygen species (ROS) induced by N-acetylchitooligosaccharide elicitor in rice cells was associated with the activation of phopholipase C (PLC) and phospholipase D (PLD). The activation of both enzymes was observed for the first phase of ROS generation but only the activation of PLD was evident for the second response. Activation of PLD was associated with its recruitment to the membrane. Enzymatic products of these phospholipases, diacylglycerol (DG) and phosphatidic acid (PA), could induce ROS generation by themselves. Moreover, the addition of these lipids compensated the inhibition of the second phase of ROS generation by cycloheximide, indicating the involvement of the synthesis of PLD or related proteins in the second phase of ROS generation. DG and PA also induced the expression of elicitor-responsive genes in the absence of the elicitor. They could not induce phytoalexin biosynthesis by themselves but greatly enhanced the elicitor-induced phytoalexin accumulation. Further, the inhibition of PLD by 1-butanol inhibited the elicitor-induced phytoalexin accumulation, indicating the involvement of PLD and its reaction product, PA, in the induction of phytoalexin biosynthesis. These results indicated the importance of phospholipid signaling, especially by PLD and its product PA, in plant defense Biphasic generation of reactive oxygen species (ROS) responses.

Keywords: Oryza Sativa; Chitin oligosaccharide elicitor; defense response; PLC; PLD.
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