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Published on July 21, 2006; 10.1104/pp.106.080283

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Received March 12, 2006
Accepted July 10, 2006

Characterization of a Novel Plant Promoter Specifically Induced by Heavy Metal and Identification of the Promoter Regions Conferring Heavy Metal Responsiveness

Xiaoting Qi , Yuxiu Zhang , and Tuanyao Chai *

Department of Biology, Graduate University of Chinese Academy of Sciences, Yuquan road 19, Beijing 100049, P. R. China
Department of Bio-engineering, School of Chemical and Environmental Engineering, China University of Mining and Technology (Beijing), Xueyaun road 11, Beijing 100083, P. R. China

* Corresponding author; email: tychai{at}gucas.ac.cn; tuanyaochai@hotmail.com.

The bean PvSR2 gene (Phaseolus vulgaris stress-related gene no. 2) responds to heavy metals but not to other forms of environment stresses. To elucidate its heavy metal-regulatory mechanism at the transcriptional level, we isolated and characterized the promoter region (-1623/+48) of PvSR2 (P-PvSR). Deletions from the 5'-end revealed that a sequence between -222 and -147 relative to the transcriptional start site was sufficient for heavy metal-specific induction of P-PvSR. Detailed analysis of this 76 bp fragment indicated that heavy metal-responsive elements (HMREs) were localized in two regions (-222/-188 and -187/-147), each of which could separately confer heavy metal-responsive expression on the {beta}-glucuronidase (GUS) gene in the context of a minimal CaMV 35S promoter. Region I (-222/-188) contains a motif (MRE-like sequence) similar to the consensus metal-regulatory element (MRE) of the animal metallothionein gene, and mutation of this motif eliminated the heavy metal-inducible function of region I. Region II (-187/-147) had no similarity to previously identified cis-acting elements involved in heavy metal induction, suggesting the presence of a novel HMRE. Transformed tobacco seedlings expressing GUS under control of the PvSR2 promoter region (-687/+48) showed heavy metal-specific responsive activity which depended on the type and concentration of the heavy metal and the type of organ. These findings further our understanding of the regulation of PvSR2 expression and provide a new heavy-metal-inducible promoter system in transgenic plants.




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