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JXB Advance Access originally published online on November 16, 2006
Journal of Experimental Botany 2007 58(4):765-772; doi:10.1093/jxb/erl210
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© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Approaches to Cellular Imaging

3-D cell-level chlorophyll fluorescence imaging of ozone-injured sunflower leaves using a new passive light microscope system

Ryosuke Endo1,* and Kenji Omasa2

1Advanced Research Institute for the Sciences and Humanities, Nihon University, 6F Ichigaya Tokyu Building, 4-2-1 Kudan-kita, Chiyoda, Tokyo 102-0073, Japan
2Department of Biological and Environmental Engineering, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo, Tokyo 113-8657, Japan

* To whom correspondence should be addressed. E-mail: ryosuke-endo{at}arish.nihon-u.ac.jp

A passive light microscope system has been developed, capable of reconstructing an extended-focus 3-D cell-level image of chlorophyll fluorescence and {Phi}PSII of intact attached leaves using a limited number of focal plane images of chlorophyll fluorescence. Using this system, the relationships between the depth of the mesophyll cells in spongy tissue and the intensity of the chlorophyll fluorescence and the {Phi}PSII were investigated in sunflower leaves exposed to 300 ppb ozone for 12 h at a PPFD of 300 µmol m–2 s–1 actinic light. After ozone exposure, fluorescence intensity (F) largely decreased in the cells just under the epidermal cells (within ~20 µm of the epidermal cells), but the sites where fluorescence intensity decreased had no relationship to the position of the stomata. By contrast, the distribution of {Phi}PSII showed no change after the ozone exposure. These findings suggest that ozone-induced inhibition occurs in the cells just under the epidermal cells by reducing the light absorption of the chloroplasts, while the operating quantum efficiency of PSII photochemistry is maintained.

Key words: Chlorophyll fluorescence, 3-D imaging, microscope, ozone

Received 8 May 2006; Accepted 13 September 2006


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