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Published on January 27, 2006; 10.1104/pp.105.071993

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Received September 29, 2005
Returned for revision November 14, 2005
Accepted January 12, 2006

Effects of feeding Spodoptera littoralis on Lima bean leaves. III. Membrane Depolarization and Involvement of Hydrogen Peroxide

Massimo E. Maffei *, Axel Mithöfer , Gen-ichiro Arimura , Hannes Uchtenhagen , Simone Bossi , Cinzia M. Bertea , Laura Starvaggi Cucuzza , Mara Novero , Veronica Volpe , Stefano Quadro , and Wilhelm Boland

Department of Plant Biology and Centre of Excellence CEBIOVEM, University of Turin, Viale P.A. Mattioli 25, I-10125 Turin, Italy
Max Planck Institut for Chemical Ecology, Bioorganic Chemistry, Hans-Knöll-Str. 8, D-07745 Jena, Germany
Max Planck Institut for Chemical Ecology, Bioorganic Chemistry, Hans-Knöll-Str. 8, D-07745 Jena, Germany

* Corresponding author; email: massimo.maffei{at}unito.it.

In response to herbivore (Spodoptera littoralis) attack, Lima bean (Phaseolus lunatus L.) leaves produced H2O2, in concentrations which were higher when compared to mechanically damaged leaves. Cellular and subcellular localization analyses revealed that H2O2 was mainly localized in mechanically damaged (MD) and herbivore wounded (HW) zones and spread throughout the veins and tissues. Preferentially, H2O2 was found in cell walls of spongy and mesophyll cells facing intercellular spaces, even though confocal laser scanning microscopy (CLSM) analyses also revealed the presence of H2O2 in mitochondria/peroxisomes. Increased gene and enzyme activations of superoxide dismutase after HW were in agreement with CLSM data. After MD, additional application of H2O2 prompted a transient transmembrane potential (Vm) depolarization, with a Vm depolarization rate that was higher when compared to HW leaves. In transgenic soybean suspension cells expressing the Ca2+ sensing aequorin system, increasing amounts of added H2O2 correlated with a higher cytosolic calcium ([Ca2+]cyt) concentration. In MD and HW leaves, H2O2 also triggered the increase of [Ca2+]cyt, but MD-elicited [Ca2+]cyt increase was more pronounced when compared to HW leaves after addition of exogenous H2O2. The results clearly indicate that Vm depolarization caused by HW makes the membrane potential more positive and reduces the ability of Lima bean leaves to react to signaling molecules.




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