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Plant and Cell Physiology Advance Access published online on May 26, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci135
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received January 28, 2005
Accepted May 19, 2005

Regular Paper

Acclimation to Diverse Environmental Stresses Caused by a Suppression of Cytosolic Ascorbate Peroxidase in Tobacco BY-2 Cells

Takahiro Ishikawa 1*, Yukari Morimoto 1, Rapolu Madhusudhan 1, Yoshihiro Sawa 1, Hitoshi Shibata 1, Yukinori Yabuta 2, Ayako Nishizawa 2, and Shigeru Shigeoka 2

1 Faculty of Life and Environmental Science, Shimane University, 1060 Nishikawatsu, Matsue 690-8504, Shimane, Japan
2 Department of Advanced Bioscience, Kinki University, 3327-204 Nakamachi, Nara 631-8505, Nara, Japan

* To whom correspondence should be addressed.
Takahiro Ishikawa, E-mail: ishikawa{at}life.shimane-u.ac.jp


  Abstract

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 cosuppression by expression of Arabidopsis APX1 cDNA under the 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 upregulated 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 MAP kinase of approximately 46 kDa was predominantly active in the APX-suppressed cells and transcript levels of both NPK1 and NDPK2 were upregulated. 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.

Keywords: Active oxygen species; Ascorbate peroxidase; Diverse stress tolerance; Redox; Tobacco BY-2.
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