First published online November 7, 2002; 10.1104/pp.008243
Plant Physiol, December 2002, Vol. 130, pp. 1927-1937
Control of Demand-Driven Biosynthesis of Glutathione in Green
Arabidopsis Suspension Culture Cells1
Andreas J.
Meyer2* and
Mark D.
Fricker
Department of Plant Sciences, University of Oxford, South Parks
Road, Oxford OX1 3RB, United Kingdom
We have investigated what limits demand-driven de novo glutathione
(GSH) biosynthesis in green Arabidopsis suspension culture cells. GSH
is the most abundant low-molecular weight thiol in most plants and can
be quantified using monochlorobimane to fluorescently label GSH in live
cells. Progress curves for labeling reached a plateau as all the
cytoplasmic GSH was conjugated. In the presence of excess
monochlorobimane, a second, almost linear phase of labeling was
observed, after a lag of 2 to 3 h, that was then maintained for an
extended period. The increase in fluorescence was shown to be because
of de novo GSH biosynthesis by high-performance liquid chromatography
analysis and was eliminated by
DL-buthionine-[S,R]-sulfoximine, a
specific inhibitor of GSH biosynthesis, or reduced by inhibitors of
transcription and translation. The rate of GSH biosynthesis during the
linear phase was 8.9 ± 1.4 nmol g fresh weight 1
min1 and was not affected by addition of glutamate,
glycine, or cysteine, the immediate precursors needed for GSH
biosynthesis. Likewise, the synthesis rate was not affected by
pretreatment with aminotriazole, menadione, jasmonic acid, or cadmium,
all of which cause oxidative stress and up-regulate expression of GSH
biosynthetic genes. The lag phase was markedly reduced by aminotriazole
and menadione and marginally by jasmonic acid, suggesting the system
was primed to react faster after mild stress. In contrast to the other
feeding experiments, exclusion of SO42 from
the medium abolished the second phase completely. This suggests demand-driven GSH biosynthesis is directly coupled to uptake of SO42 and that the linear increase in
fluorescence reflects flux through the entire
SO42 assimilation pathway.
1
This work was partially supported by Aventis
Crop Science Ltd.
2
Present address: Institut für Forstbotanik
und Baumphysiologie, Professur für Baumphysiologie,
Universität Freiburg, Georges-Köhler-Allee 053, D-79085 Freiburg, Germany.
*
Corresponding author; e-mail Andreas.Meyer{at}sonne.uni-freiburg.de;
fax 49-761-2038302.
© 2002 American Society of Plant Biologists
This article has been cited by other articles:
|
|
|
|
 
G. Queval, D. Thominet, H. Vanacker, M. Miginiac-Maslow, B. Gakiere, and G. Noctor
H2O2-Activated Up-Regulation of Glutathione in Arabidopsis Involves Induction of Genes Encoding Enzymes Involved in Cysteine Synthesis in the Chloroplast
Mol Plant,
March 1, 2009;
2(2):
344 - 356.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. M. Hicks, R. E. Cahoon, E. R. Bonner, R. S. Rivard, J. Sheffield, and J. M. Jez
Thiol-Based Regulation of Redox-Active Glutamate-Cysteine Ligase from Arabidopsis thaliana
PLANT CELL,
August 1, 2007;
19(8):
2653 - 2661.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Wirtz and R. Hell
Dominant-Negative Modification Reveals the Regulatory Function of the Multimeric Cysteine Synthase Protein Complex in Transgenic Tobacco
PLANT CELL,
February 1, 2007;
19(2):
625 - 639.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. G. Cairns, M. Pasternak, A. Wachter, C. S. Cobbett, and A. J. Meyer
Maturation of Arabidopsis Seeds Is Dependent on Glutathione Biosynthesis within the Embryo
Plant Physiology,
June 1, 2006;
141(2):
446 - 455.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D.-Y. Kim, L. Bovet, S. Kushnir, E. W. Noh, E. Martinoia, and Y. Lee
AtATM3 Is Involved in Heavy Metal Resistance in Arabidopsis
Plant Physiology,
March 1, 2006;
140(3):
922 - 932.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Ortega-Villasante, R. Rellan-Alvarez, F. F. Del Campo, R. O. Carpena-Ruiz, and L. E. Hernandez
Cellular damage induced by cadmium and mercury in Medicago sativa
J. Exp. Bot.,
August 1, 2005;
56(418):
2239 - 2251.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. P. Mezzari, K. Walters, M. Jelinkova, M.-C. Shih, C. L. Just, and J. L. Schnoor
Gene Expression and Microscopic Analysis of Arabidopsis Exposed to Chloroacetanilide Herbicides and Explosive Compounds. A Phytoremediation Approach
Plant Physiology,
June 1, 2005;
138(2):
858 - 869.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Ball, G.-P. Accotto, U. Bechtold, G. Creissen, D. Funck, A. Jimenez, B. Kular, N. Leyland, J. Mejia-Carranza, H. Reynolds, et al.
Evidence for a Direct Link between Glutathione Biosynthesis and Stress Defense Gene Expression in Arabidopsis
PLANT CELL,
September 1, 2004;
16(9):
2448 - 2462.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
