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Plant Physiol, January 2002, Vol. 128, pp. 95-107

Constitutive Overexpression of Cystathionine gamma -Synthase in Arabidopsis Leads to Accumulation of Soluble Methionine and S-Methylmethionine1

Jungsup Kim,23 Minsang Lee,2 Radhika Chalam, Melinda Neal Martin, Thomas Leustek,* and Wout Boerjan

Biotechnology Center for Agriculture and the Environment, Plant Science Department, Rutgers University, New Brunswick, New Jersey 08901-8520 (J.K., M.L., R.C., M.N.M., T.L.); and Department of Plant Genetics, Vlaams Interuniversitair Instituut voor Biotechnologie, Universiteit Gent, K.L. Ledeganckstraat 35, 9000 Gent, Belgium (W.B.)

The committing step in Met and S-adenosyl-L-Met (SAM) synthesis is catalyzed by cystathionine gamma -synthase (CGS). Transgenic Arabidopsis plants overexpressing CGS under control of the cauliflower mosaic virus 35S promoter show increased soluble Met and its metabolite S-methyl-Met, but only at specific stages of development. The highest level of Met and S-methyl-Met was observed in seedling tissues and in flowers, siliques, and roots of mature plants where they accumulate 8- to 20-fold above wild type, whereas the level in mature leaves and other tissues is no greater than wild type. CGS-overexpressing seedlings are resistant to ethionine, a toxic Met analog. With these properties the transgenic lines resemble mto1, an Arabidopsis, CGS-mutant inactivated in the autogenous control mechanism for Met-dependent down-regulation of CGS expression. However, wild-type CGS was overexpressed in the transgenic plants, indicating that autogenous control can be overcome by increasing the level of CGS mRNA through transcriptional control. Several of the transgenic lines show silencing of CGS resulting in deformed plants with a reduced capacity for reproductive growth. Exogenous feeding of Met to the most severely affected plants partially restores their growth. Similar morphological deformities are observed in plants cosuppressed for SAM synthetase, even though such plants accumulate 250-fold more soluble Met than wild type and they overexpress CGS. The results suggest that the abnormalities associated with CGS and SAM synthetase silencing are due in part to a reduced ability to produce SAM and that SAM may be a regulator of CGS expression.

1 This work was supported by the National Science Foundation (grant no. MCB-9728661 to T.L. and MCB-0094062 to M.N.M.) and Pioneer Hi-Bred, Inc.

2 These authors contributed equally to the work.

3 Present address: Department of Biological Sciences, University of Delaware, Newark, DE 19711.

* Corresponding author; e-mail leustek{at}aesop.rutgers.edu; fax 732-932-0312.

© 2002 American Society of Plant Physiologists


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