Glutathione is a key antioxidant metabolite to cope with mercury and cadmium stress
Glutathione (GSH) plays a dual role under heavy metal stress, as antioxidant metabolite and as precursor of phytochelatins (PCs). Studying the responses of the GSH metabolism to heavy metals is important to improve tolerance.
We studied the oxidative stress signature of three γ-glutamylcysteine synthetase (γECS) Arabidopsis thaliana allele mutants (rax1-1, cad2-1, and pad2-1), first enzymatic step in the GSH synthetic pathway, when treated with 10 μM Cd or Hg for 72 h.
GSH concentration was lower in the mutants (45 % rax1-1; 30 % cad2-1; and 20 % pad2-1), which was also associated with inferior translocation of Cd or Hg to shoots, than in wild type Col-0. Glutathione reductase (GR) and NADPH-oxidase activities were inhibited in roots, phytotoxic effects consistently more pronounced in the mutants, particularly in pad2-1. Non-photochemical quenching augmented with exposure time to Cd or Hg in Col-0, but not so in the γECS mutants. Mercury caused severe damage in cad2-1 and pad2-1 root proteins profile; toxic effects confirmed by GR and H+-ATPase immunodetection. PCs appeared in Col-0 roots under metal stress, and surprisingly accumulated in rax1-1. γECS immunodetection revealed its overexpression in rax1-1.
A minimum amount of GSH may be required for adequate metal tolerance, where γECS expression could compensate GSH deficiency under stress.
KeywordsArabidopsis thaliana Biothiols Cadmium γ-glutamylcysteine synthetase Glutathione Mercury Phytochelatins
This work was funded by the Ministry of Economy and Competitivity (PROBIOMET AGL2010-15151), Fundación Ramón Areces, and Junta Comunidades Castilla-La Mancha (FITOALMA2, POII10-0087-6458). We are extremely grateful to Prof. Phil Mullineaux (University of Essex, UK) for his donation of rax1-1 mutant seeds. We thank the comments of two anonymous reviewers which allowed substantial improvement of the manuscript.
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