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Parallel assessment of ROS formation and photosynthesis in leaves by fluorescence imaging

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Abstract

Stress-induced generation of reactive oxygen species (ROS) leads to lowering of the biochemical yield of photosynthesis in plant leaves. The detrimental effects of oxidative stress by paraquat are initiated by the generation of superoxide anion radicals in the vicinity of the thylakoid membrane. However, direct proof of ROS production has been elusive. In this study, we report first in vivo detection and imaging of the generated superoxide in illuminated tobacco leaves following paraquat infiltration. This was done using a newly developed imaging apparatus capable of detecting changes in the fluorescence of the ROS sensor 3-(N-dansyl)aminomethyl-2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrole. Under identical conditions, the effects on photosynthesis caused by the oxidative stress were assessed via chlorophyll fluorescence imaging and the saturation pulse method. In the future, the combination of these two imaging techniques may provide information on the spatial distribution and extent of stress induced ROS production in plant leaves, as well as on the protective ability of various free radical scavengers and antioxidants.

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Abbreviations

Fm :

Maximum fluorescence yield in the dark adapted state

Fo :

Minimum fluorescence yield in the dark adapted state

F ′m :

Maximum fluorescence yield in the light adapted state

Fv/Fm :

Maximum photochemical yield of PS II in the dark adapted state

HO-1889NH:

3-(N-dansyl)aminomethyl-2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrole

LED:

Light-emitting diode

NPQ:

Non-photochemical quenching

PAM:

Pulse amplitude modulation

PAR:

Photosynthetically active radiation

Paraquat:

N,N′-Dimethyl-4,4′-bipyridinium dichloride

PS:

Photosystem

ROS:

Reactive oxygen species

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Acknowledgments

É.H. wishes to acknowledge the support of the Economic Competitiveness Operational Program (ECOP, grant number GVOP 3.2.1-2004-04-144). We thank Dr. Tamás Kálai and Prof. Kálmán Hideg (Pécs University, Department of Organic and Medicinal Chemistry, Hungary, tamas.kalai@aok.pte.hu) for the fluorescent ROS sensor HO-1889NH.

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Authors and Affiliations

  1. Institute of Plant Biology, Biological Research Center, P.O. Box 521, Szeged, 6701, Hungary

    Éva Hideg

  2. Julius-von-Sachs Institut für Biowissenschaften, Universität Würzburg, Würzburg, Germany

    Ulrich Schreiber

Authors
  1. Éva Hideg
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  2. Ulrich Schreiber
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Correspondence to Éva Hideg.

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Hideg, É., Schreiber, U. Parallel assessment of ROS formation and photosynthesis in leaves by fluorescence imaging. Photosynth Res 92, 103–108 (2007). https://doi.org/10.1007/s11120-007-9146-4

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  • DOI: https://doi.org/10.1007/s11120-007-9146-4

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