Acclimation to diverse environmental stresses caused by a suppression of cytosolic ascorbate peroxidase in tobacco BY-2 cells.
The active oxygen species (AOS) that arise from normal metabolic processes are kept under tight control by various antioxidant mechanisms. AOS are important signal molecules that regulate many physiological processes, including environmental stress responses. In this work, we have investigated the effect of lowered cytosolic ascorbate peroxidase (APX) activity in transgenic tobacco BY-2 cells, using two transformed BY-2 cell lines, cAPX-S2 and cAPX-S3, resulting from co-suppression by expression of Arabidopsis APX1 cDNA under the cauliflower mosaic virus (CaMV) 35S promoter. cAPX-S2 and cAPX-S3 possessed 50 and 75% lower cytosolic APX activity, respectively, compared with that in the untransformed cells. Chemical fluorescence analysis indicated that the AOS levels were markedly higher in the two APX-suppressed cell lines than in the wild-type cells. However, there were no substantial differences in the activity levels of the various other antioxidant enzymes. Interestingly, the APX-suppressed cells showed different responses and tolerances to environmental stresses, such as heat and salinity. Suppression subtractive hybridization revealed that several heat- and salt stress-inducible genes were up-regulated in cAPX-S3 cells. HSP70, DnaJ-like protein and purple acid phosphatase were among the constitutively induced genes. An in-gel kinase assay suggested that a mitogen-activated protein (MAP) kinase of approximately 46 kDa was predominantly active in the APX-suppressed cells, and transcript levels of both nicotiana protein kinase 1 (NPK1) and nucleoside diphosphate kinase 2 (NDPK2) were up-regulated. These data suggest the possibility that MAP kinase cascades are activated by subtle imbalances in the homeostasis of the cellular redox status caused by lowered cytosolic APX activity.
Ishikawa
Takahiro
T
Faculty of Life and Environmental Science, Shimane University, 1060 Nishikawatsu, Matsue, Shimane, 690-8504 Japan. ishikawa@life.shimane-u.ac.jp
Morimoto
Yukari
Y
Madhusudhan
Rapolu
R
Sawa
Yoshihiro
Y
Shibata
Hitoshi
H
Yabuta
Yukinori
Y
Nishizawa
Ayako
A
Shigeoka
Shigeru
S
D060387:Ascorbate Peroxidases; D001483:Base Sequence; D002460:Cell Line; D003600:Cytosol / Q000201:enzymology*; D017931:DNA Primers; D006358:Hot Temperature; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D012492:Salts; D014026:Nicotiana / Q000166:cytology / Q000201:enzymology*
D017931:DNA Primers; D012492:Salts; D010544:Peroxidases; D060387:Ascorbate Peroxidases