Activation of Barley S-Adenosylmethionine Synthetase1 Gene Promoter in Response to Phytohormones and Abiotic Stresses

  • Kim, Jae-Yoon (Plant Molecular Breeding Lab., Division of Biotechnology, Korea University) ;
  • Kim, Dae-Yeon (Plant Molecular Breeding Lab., Division of Biotechnology, Korea University) ;
  • Jung, Je-Hyeong (Plant Molecular Breeding Lab., Division of Biotechnology, Korea University) ;
  • Hong, Min-Jeong (Plant Molecular Breeding Lab., Division of Biotechnology, Korea University) ;
  • Heo, Hwa-Young (Rural Development Administration(RDA)) ;
  • Johnson, Jerry W. (Department of Crop and Soil Science, University of Georgia Griffin Campus) ;
  • Kim, Tae-Ho (BioGreen 21 Program, Rural Development Administration) ;
  • Seo, Yong-Weon (Plant Molecular Breeding Lab., Division of Biotechnology, Korea University)
  • Published : 2007.03.31

Abstract

Barley S-adenosylmethionine synthetase1 gene, which was differentially expressed in seed development of extra early barley, was regulated by the phytohormones and abiotic stresses. In order to identify the regulation regions which were involved in transcriptional control of the phytohormones and abiotic stresses, we isolated 1459 bp fragment of HvSAMS1 gene promoter using genome walking strategy and deletion series were constructed. Deleted upstream fragments(-1459, -1223, -999, -766, -545, -301 bp) were fused to the GUS reporter gene and evaluated via Agrobacterium-mediated transient expression assay. Increased GUS activity of HvSMAS1 promoter -301/GUS construct under each of NaCl, $GA_3$, ABA and ethylene application was found. However, GUS activity was negligible in the leaves transformed with the HvSMAS1 promoter(-1459, -1223, -999, -766 and -545)/GUS constructs. No significant induction of GUS activity was observed for the ethionine and spermidine treatments. In order to locate promoter sequence of the HvSAMS1 gene that was critical for the activation of gene expression, deletion and addition promoter derivatives(+, includes 43 bp of 5' ORF) of the HvSAMS1 gene fused to the GUS reporter gene were applied. The tobacco leaves which harbored the additional HvSAMS1 promoter(-1459+, -1459 to -546, -545+ and -301+)/GUS construct did not significantly induce GUS activity as compared to the HvSAMS1 promoter(-1459, -545 and -301)/GUS constructs under each of NaCl, ABA and $GA_3$ treatment. However, the GUS activity was high in the tobacco leaves which harboring the -211 to -141 regions of the HvSAMS1 promoter. This result suggested that HvSAMS1 gene expression might be regulated by this region(from -211 to -141).