Distinct cis-elements in the Asparagus officinalis asparagine synthetase promoter respond to carbohydrate and senescence signals
Somrutai Winichayakul A , Richard L. Moyle A C , Dacey J. Ryan B , Kevin J. F. Farnden A , Kevin M. Davies B E and Simon A. Coupe B DA Department of Biochemistry, University of Otago, PO Box 56, Dunedin, NZ.
B New Zealand Institute for Crop and Food Research Limited, Private Bag 11-600, Palmerston North, NZ.
C Current address: Department of Botany, John Hines Building, The University of Queensland, Brisbane, Qld 4072, Australia.
D Current address: Department of Animal and Plant Sciences, University of Sheffield, Western Bank, Sheffield, S10 2TN, UK.
E Corresponding author; email: daviesk@crop.cri.nz
Functional Plant Biology 31(6) 573-582 https://doi.org/10.1071/FP03198
Submitted: 21 October 2003 Accepted: 19 February 2004 Published: 23 June 2004
Abstract
The Asparagus officinalis L. asparagine (Asn) synthetase (AS) promoter was analysed for elements responding to carbohydrate and senescence signals. Transgenic Arabidopsis thaliana L. plants containing deletion constructs of the –1958 bp AS promoter linked to the β-glucuronidase (GUS) reporter gene (AS::GUS) were analysed by measuring GUS specific activity. Inclusion of sucrose (Suc), glucose (Glc) or fructose (Fru) in plant media repressed levels of GUS activity in –1958AS::GUS plants, regardless of the light environment, with increases in GUS found 1 d after incubation on Suc-lacking media. Hexokinase is likely to be involved in the signal pathway, as Suc, Glc, Fru, 2-deoxy-d-glucose and mannose were more effective repressors than 3-O-methylglucose, and the hexokinase inhibitor mannoheptulose reduced repression. Plants containing AS::GUS constructs with deletions that reduced the promoter to less than –405 bp did not show low sugar induction. AS::GUS activity was significantly higher in excised leaves induced to senesce by dark storage for 24 h, compared to fresh leaves, for lines containing at least –640 bp of the AS promoter but not those with –523 bp or smaller promoter fragments. Fusion of the –640 to –523 bp region to a –381AS::GUS construct generated a promoter that retained senescence induction but lacked low sugar induction. Alignment of this region to the 33-bp senescence-related sequence of the Arabidopsis and Brassica napus L. SAG12 promoters identified the sequence TTGCACG as being conserved in all the promoters, and which may be an important senescence-responsive element.
Keywords: asparagine synthetase, asparagus, gene regulation, promoter, senescence, sugar.
Acknowledgments
We thank Ian King for care of Arabidopsis transgenics. The New Zealand Foundation for Research, Science and Technology supported this work financially and SW was partially funded by a University of Otago 125th Jubilee Scholarship.
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