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Title of Journal: Plant Mol Biol

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Abbravation: Plant Molecular Biology

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Springer Netherlands

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DOI

10.1007/s10333-006-0064-4

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ISSN

1573-5028

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Overproduction of stromal ferredoxinNADPH oxidore

Authors: Rajesh Kumar Tewari Mamoru Satoh Sayaka Kado Kohei Mishina Misato Anma Kazuhiko Enami Mitsumasa Hanaoka Masami Watanabe
Publish Date: 2014/09/26
Volume: 86, Issue: 6, Pages: 627-639
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Abstract

The isolation of Brassica napus leaf protoplasts induces reactive oxygen species generation and accumulation in the chloroplasts An activated isoform of NADPH oxidaselike protein was detected in the protoplasts and the protoplast chloroplasts The purpose of this study is to define the NADH oxidaselike activities in the H2O2accumulating protoplast chloroplasts Proteomic analysis of this protein revealed an isoform of ferredoxinNADPH oxidoreductase FNR1 While leaves highly expressed the LFNR1 transcript protoplasts decreased the expression significantly The protoplast chloroplasts predominantly expressed soluble FNR1 proteins While the albino leaves of white kale Brassica oleracea var acephala f tricolor cv white pigeon expressed FNR1 protein at the same level as B napus leaves the protoplasts of albino leaves displayed reduced FNR1 expression The albino leaf protoplasts of white kale generated and accumulated H2O2 in the cytoplasm and on the plasma membrane Intracellular pH showed that the chloroplasts were acidic which suggest that excess H+ was generated in chloroplast stroma NADPH content of the protoplast chloroplasts increased by over sixfold during the isolation of protoplasts This study reports a possibility of mediating electrons to oxygen by an overproduced soluble FNR and suggests that the FNR has a function in utilizing any excess reducing power of NADPHThe authors are grateful to Prof Poul Erik Jensen University of Copenhagen and Prof Toshiraru Hase Institute for Protein Research Osaka University for kindly gifting FNR1 antibody This study was supported by the Japan Society for the Promotion of Science JSPS GrantinAid for a postdoctoral fellowship P08413 to RKT and The Sumitomo Foundation to MW

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