Glutathione reductase activity and isoforms in leaves and roots of wheat plants subjected to cadmium stress.
The behavior of glutathione reductase (GR, EC 1.6.4.2) activity and isoforms were analyzed in wheat (Triticum aestivum L.) leaves and roots exposed to a chronic treatment with a toxic cadmium (Cd) concentration. A significant growth inhibition (up to 55%) was found in leaves at 7, 14 and 21 days, whereas roots were affected (51%) only after three weeks. Wheat plants grown in the presence of 100microM Cd showed a time-dependent accumulation of this metal, with Cd concentration being 10-fold higher in roots than in leaves. Nevertheless, lipid peroxidation was augmented in leaves in all experiments, but not in roots until 21 days. Cadmium treatment altered neither the GR activity nor the isoform pattern in the leaves. However, GR activity increased 111% and 200% in roots at 7 and 14 days, respectively, returning to control levels after 21 days. Three GR isoforms were found in roots of control and treated plants, two of which were enhanced by Cd treatment at 7 and 14 days, as assessed by activity staining on native gels. The changes in the isoform pattern modified the global kinetic properties of GR, thereby decreasing significantly (2.5-fold) the Michaelis constant (K(m)) value for oxidized glutathione. Isozyme induction was not associated with an enhancement of GR mRNA and protein expression, indicating that post-translational modification could occur. Our data demonstrated that up-regulation of GR activity by the induction of distinctive isoforms occurs as a defense mechanism against Cd-generated oxidative stress in roots.
Yannarelli
Gustavo G
GG
Departamento de Química Biológica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956, (1113) Buenos Aires, Argentina. gyanna@ffyb.uba.ar
Fernández-Alvarez
Ana J
AJ
Santa-Cruz
Diego M
DM
Tomaro
María L
ML
D002104:Cadmium / Q000633:toxicity*; D017931:DNA Primers; D005980:Glutathione Reductase / Q000235:genetics / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D018515:Plant Leaves / Q000187:drug effects / Q000201:enzymology*; D010940:Plant Proteins / Q000378:metabolism; D018517:Plant Roots / Q000187:drug effects / Q000201:enzymology*; D012333:RNA, Messenger / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014908:Triticum / Q000187:drug effects / Q000201:enzymology*
D017931:DNA Primers; D007527:Isoenzymes; D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D002104:Cadmium; D005980:Glutathione Reductase