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Plant and Cell Physiology Advance Access published online on December 7, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci235
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received August 29, 2005
Accepted November 25, 2005

Regular Paper

Photoinactivation of Ascorbate Peroxidase in Isolated Tobacco Chloroplasts: Galdieria partita APX Maintains the Electron Flux through the Water-Water Cycle in Transplastomic Tobacco Plants

Chikahiro Miyake 1 *, Yuki Shinzaki 1, Minori Nishioka 1, Sayaka Horiguchi 1, and Ken-Ichi Tomizawa 1

1 Research Institute of Innovative Technology for the Earth (RITE), 9-2 Kizugawadai, Kizu-cho, Soraku-gun, Kyoto 619-0292 Japan

* To whom correspondence should be addressed.
Chikahiro Miyake, E-mail: cmiyake{at}rite.or.jp


  Abstract

We evaluated the H2O2 scavenging activity of the water-water cycle in illuminated intact chloroplasts isolated from tobacco leaves. Illumination under conditions that limited photosynthesis (red light (>640nm), 250 µmol photons m-2 s-1 in the absence of HCO3-) caused chloroplasts to take up O2 and accumulate H2O2. Concomitant with the O2-uptake, both APX and GAPDH lost their activities. However, SOD, MDAR, DHAR, and GR activities remained unaffected. The extent to which the photosynthetic linear electron flow decreased was small compared to the decline in APX activity. Therefore, the loss of APX activity lowered the electron flux through the WWC, as evidenced by a decrease in {Phi}(PSII)xPFD. To verify these interpretations, we created a transplastomic tobacco line in which an H2O2-insensitive APX from the red alga, Galdieria partita, was overproduced in the chloroplasts. In intact transplastomic chloroplasts which were illuminated under conditions that limited photosynthesis, neither O2-uptake nor H2O2-accumulation occurred. Furthermore, the electron flux through the WWC and the activity of GAPDH were maintained. The present work is the first report of APX inactivation by endogenous H2O2 in intact chloroplasts.

Keywords: Active oxygen; Ascorbate peroxidase (APX); Chloroplasts; Hydrogen peroxide; The Water-Water Cycle; Transplastomic Tobacco.
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