JXB Advance Access originally published online on March 12, 2004
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Journal of Experimental Botany, Vol. 55, No. 398, pp. 837-845, April 1, 2004
© 2004 Oxford University Press
Cell and Molecular Biology, Biochemistry and Molecular Physiology |
Regulation of sulphate assimilation by glutathione in poplars (Populus tremulaxP. alba) of wild type and overexpressing
-glutamylcysteine synthetase in the cytosol
Received 17 October 2003; Accepted 26 December 2003
1 Albert-Ludwigs-University of Freiburg, Institute of Forest Botany and Tree Physiology, Georges-Köhler-Allee 053, D-79110 Freiburg, Germany
2 Heidelberger Institute of Plant Sciences (HIP), Im Neuenheimer Feld 360, D-69120 Heidelberg, Germany
* To whom the correspondence should be addressed. Fax: +49 761 2038302. E-mail: Stanislav.Kopriva{at}ctp.uni-freiburg.de
Abbreviations: APR, adenosine 5'-phosphosulphate reductase; APS, adenosine 5'-phosphosulphate; ATPS, ATP sulphurylase; GSH, glutathione; GSHS, glutathione synthetase;
-ECS,
-glutamylcysteine synthetase; OAS, O-acetylserine; OASTL, O-acetylserine (thiol)lyase; SAT, serine acetyltransferase; SIR, sulphite reductase.
Glutathione (GSH) is the major low molecular weight thiol in plants with different functions in stress defence and the transport and storage of sulphur. Its synthesis is dependent on the supply of its constituent amino acids cysteine, glutamate, and glycine. GSH is a feedback inhibitor of the sulphate assimilation pathway, the primary source of cysteine synthesis. Sulphate assimilation has been analysed in transgenic poplars (Populus tremulaxP. alba) overexpressing
-glutamylcysteine synthetase, the key enzyme of GSH synthesis, and the results compared with the effects of exogenously added GSH. Although foliar GSH levels were 34-fold increased in the transgenic plants, the activities of enzymes of sulphate assimilation, namely ATP sulphurylase, adenosine 5'-phosphosulphate reductase (APR), sulphite reductase, serine acetyltransferase, and O-acetylserine (thiol)lyase were not affected in three transgenic lines compared with the wild type. Also the mRNA levels of these enzymes were not altered by the increased GSH levels. By contrast, an increase in GSH content due to exogenously supplied GSH resulted in a strong reduction in APR activity and mRNA accumulation. This feedback regulation was reverted by simultaneous addition of O-acetylserine (OAS). However, OAS measurements revealed that OAS cannot be the only signal responsible for the lack of feedback regulation of APR by GSH in the transgenic poplars.
Key words: Adenosine 5'-phosphosulphate reductase, cysteine synthesis, glutathione, poplar, sulphate assimilation.
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