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Published on February 17, 2006; 10.1104/pp.105.070474

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Received August 25, 2005
Returned for revision December 4, 2005
Accepted February 6, 2006

The Role of H2O2-Producing and H2O2-Consuming Peroxidases in the Leaf Apoplast of Vigna unguiculata L. in Manganese Tolerance

Marion Maria Fecht-Christoffers , Hendrik Führs , Hans-Peter Braun , and Walter Johannes Horst *

Institute of Plant Nutrition, University of Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany
Department of Applied Genetics, University of Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany

* Corresponding author; email: horst{at}pflern.uni-hannover.de.

The apoplast is considered the leaf compartment decisive for Mn toxicity and tolerance in cowpea. Particularly apoplastic peroxidases (PODs) were proposed to be key enzymes in Mn toxicity-induced processes. The presented work focuses on the characterization of the role of H2O2-producing (NADH-peroxidase) and H2O2-consuming peroxidase (guaiacol-POD) in the apoplastic washing fluid (AWF) of leaves for early stages of Mn toxicity and genotypic differences in Mn tolerance of cowpea. Leaf AWF of the Mn-sensitive cultivar (cv) TVu 91 but not of the Mn-tolerant cv 1987 showed an increase of guaiacol-POD and NADH-peroxidase activities at elevated AWF Mn concentrations. 2D-resolutions of AWF proteins revealed that cv TVu 91 expressed more and additional proteins at high Mn treatment, whereas Mn-tolerant cv TVu 1987 remained nearly unaffected. In both cultivars, NADH-peroxidase activity and accompanied H2O2 formation rate in vitro were significantly affected by Mn2+, p-coumaric acid, and metabolites occurring in the AWF. The total phenol concentration in the AWF was indicative of advanced stages of Mn toxicity but was rather unrelated to early stages of Mn toxicity and genotypic differences in Mn tolerance. The NADH oxidation by AWF PODs was significantly delayed or enhanced in the presence of the protein-free AWF from cv TVu 1987 or cv TVu 91, respectively. HPLC analysis of AWF indicates the presence of phenols in cv TVu 1987 not observed in cv TVu 91. We conclude from our studies that the H2O2-producing NADH-peroxidase and its modulation by stimulating or inhibiting phenolic compounds in the leaf apoplast play a major role for Mn toxicity and Mn tolerance in cowpea.




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