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Plant and Cell Physiology Advance Access originally published online on November 5, 2008
Plant and Cell Physiology 2008 49(12):1879-1886; doi:10.1093/pcp/pcn170
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Imaging of NPQ and ROS Formation in Tobacco Leaves: Heat Inactivation of the Water–Water Cycle Prevents Down-Regulation of PSII

Éva Hideg1,*, Péter B. Kós1 and Ulrich Schreiber2

1Institute of Plant Biology, Biological Research Center, Szeged, 6701 Hungary
2Julius-von-Sachs Institut für Biowissenschaften, Universität Würzburg, D-97070 Germany

*Corresponding author: E-mail, ehideg{at}brc.hu; Fax, +36-62-433-434.


  Abstract

Non-photochemical chlorophyll fluorescence quenching (NPQ) plays a major role in the protection of the photosynthetic apparatus against damage by excess light, which is closely linked to the production of reactive oxygen species (ROS). The effect of a short heat treatment on NPQ and ROS production was studied with detached tobacco leaves by fluorescence imaging of chlorophyll and of the ROS sensor dye HO-1889NH. NPQ was stimulated >3-fold by 3 min pre-treatment at 44°C, in parallel with suppression of CO2 uptake, while no ROS formation could be detected. In contrast, after 3 min pre-treatment at 46°C, NPQ was suppressed and ROS formation was indicated by quenching of HO-1889NH fluorescence. After 3 min pre-treatment at 46°C and above, partial inactivation of ascorbate peroxidase and light-driven accumulation of H2O2 was also observed. These data are discussed as evidence for a decisive role of the Mehler ascorbate peroxidase or water–water cycle in the formation of the NPQ that reflects down-regulation of PSII.

Keywords: Fluorescence imaging - Heat stress-reactive oxygen species (ROS) - Mehler ascorbate peroxidase (MAP) cycle - PAM fluorometry - Photosynthetic electron transport

Abbreviations: APX, ascorbate peroxidase (EC 1.11.1.11); DAB, diamino benzidine tetrahydrochloride; Fm, maximum fluorescence yield in the dark-acclimated state; Fo, minimum fluorescence yield in the dark-acclimated state; Fm', maximum fluorescence yield in the light-acclimated state; Fv/Fm, maximum photochemical yield of PSII in the dark-acclimated state; HO-1889NH, 3-(N-dansyl)aminomethyl-2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrole; LED, light-emitting diode; MAP cycle, Mehler ascorbate peroxidase cycle; MDA, monodehydroascorbate; MDAR, monodehydroascorbate reductase; NPQ, non-photochemical quenching; PAM, pulse amplitude modulation; PAR, photosynthetically active radiation; ROS, reactive oxygen species; Y(II), photochemical yield

(Received September 5, 2008; Accepted October 31, 2008)
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