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Plant Physiol, August 2000, Vol. 123, pp. 1337-1350

Targeted Inactivation of the Plastid ndhB Gene in Tobacco Results in an Enhanced Sensitivity of Photosynthesis to Moderate Stomatal Closure¹

Eva M. Horváth, Stefan O. Peter,² Thierry Joët, Dominique Rumeau, Laurent Cournac, Gabor V. Horváth, Tony A. Kavanagh, Christian Schäfer,³ Gilles Peltier, and Peter Medgyesy^*

Biological Research Center, Hungarian Academy of Sciences, P.O. Box 521, H-6701 Szeged, Hungary (E.M.H., G.V.H., P.M.); Botanisches Institut, Universität Bayreuth, D-95440 Bayreuth, Germany (S.O.P., C.S.); Commissariat à l'Energie Atomique Cadarache, Département d'Ecophysiologie Végétale et Microbiologie, Laboratoire d'Ecophysiologie de la Photosynthèse, F-13108 Saint-Paul-lez-Durance, France (T.J., D.R., L.C., G.P.); and Department of Genetics, Trinity College, University of Dublin, Dublin 2, Ireland (T.A.K.)

The ndh genes encoding for the subunits of NAD(P)H dehydrogenase complex represent the largest family of plastid genes without a clearly defined function. Tobacco (Nicotiana tabacum) plastid transformants were produced in which the ndhB gene was inactivated by replacing it with a mutant version possessing translational stops in the coding region. Western-blot analysis indicated that no functional NAD(P)H dehydrogenase complex can be assembled in the plastid transformants. Chlorophyll fluorescence measurements showed that dark reduction of the plastoquinone pool by stromal reductants was impaired in ndhB-inactivated plants. Both the phenotype and photosynthetic performance of the plastid transformants was completely normal under favorable conditions. However, an enhanced growth retardation of ndhB-inactivated plants was revealed under humidity stress conditions causing a moderate decline in photosynthesis via stomatal closure. This distinctive phenotype was mimicked under normal humidity by spraying plants with abscisic acid. Measurements of CO₂ fixation demonstrated an enhanced decline in photosynthesis in the mutant plants under humidity stress, which could be restored to wild-type levels by elevating the external CO₂ concentration. These results suggest that the plastid NAD(P)H:plastoquinone oxidoreductase in tobacco performs a significant physiological role by facilitating photosynthesis at moderate CO₂ limitation.

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¹ This work was supported by the Országos Mszaki Fejlesztési Bizottság (no. EU-98-D8-11), the Országos Tudományos Kutatási Alap (nos. T016995 and T019759), the Volkswagen-Stiftung (no. I70961), the French-Hungarian Intergovernmental S&T Cooperation (no. F/8-95), and the European Community Biotechnology Program (no. Bio-4-97-2245). S.O.P. was the recipient of a fellowship from the Daimler-Benz-Stiftung.

² Present address: Institute of Plant Sciences, Eidgenössische Technische Hochschule Center LFW C33, Universitätstrasse 2, CH-8092 Zurich, Switzerland.

³ Present address: Molekulare Pflanzenphysiologie, Fachbereich 2: Biologie/Chemie, Universität Bremen, Postfach 33 04 40, D-28334 Bremen, Germany.

^* Corresponding author; e-mail pmedgyesy{at}matavnet.hu; fax 36-62-433434.

© 2000 American Society of Plant Physiologists

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