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H 2 O 2 in plant peroxisomes: an in vivo analysis uncovers a Ca 2+ ‐dependent scavenging system
Author(s) -
Costa Alex,
Drago Ilaria,
Behera Smrutisanjita,
Zottini Michela,
Pizzo Paola,
Schroeder Julian I.,
Pozzan Tullio,
Schiavo Fiorella Lo
Publication year - 2010
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2010.04190.x
Subject(s) - peroxisome , reactive oxygen species , arabidopsis , microbiology and biotechnology , cytoplasm , catalase , green fluorescent protein , plant cell , biochemistry , biology , in vivo , subcellular localization , oxidative stress , chemistry , gene , mutant
Summary Oxidative stress is a major challenge for all cells living in an oxygen‐based world. Among reactive oxygen species, H 2 O 2 , is a well known toxic molecule and, nowadays, considered a specific component of several signalling pathways. In order to gain insight into the roles played by H 2 O 2 in plant cells, it is necessary to have a reliable, specific and non‐invasive methodology for its in vivo detection. Hence, the genetically encoded H 2 O 2 sensor HyPer was expressed in plant cells in different subcellular compartments such as cytoplasm and peroxisomes. Moreover, with the use of the new green fluorescent protein (GFP)‐based Cameleon Ca 2+ indicator, D3cpv–KVK–SKL, targeted to peroxisomes, we demonstrated that the induction of cytoplasmic Ca 2+ increase is followed by Ca 2+ rise in the peroxisomal lumen. The analyses of HyPer fluorescence ratios were performed in leaf peroxisomes of tobacco and pre‐ and post‐bolting Arabidopsis plants. These analyses allowed us to demonstrate that an intraperoxisomal Ca 2+ rise in vivo stimulates catalase activity, increasing peroxisomal H 2 O 2 scavenging efficiency.