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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.
1
This work was supported by the Országos
M szaki 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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