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Turkish Journal of Biology

DOI

10.3906/biy-1611-56

Abstract

Cold environmental conditions influence the growth and development of plants, causing crop reduction or even plant death. Under stress conditions, cold-inducible promoters regulate cold-related gene expression as a molecular switch. Recent studies have shown that the chloroplast-expressed GPAT gene plays an important role in determining cold sensitivity. However, the mechanism of the transcriptional regulation of GPAT is ambiguous. The 5'-flanking region of GPAT with length of 1494 bp was successfully obtained by chromosome walking from Lilium pensylvanicum. The cis-elements of GPAT promoters were predicted and analyzed by a plant cis-acting regulatory DNA element database. There exist core promoter regions including TATA-box and CAAT-box and transcription regulation regions, which involve some regulatory elements such as I-box, W-box, MYB, MYC, and DREB. Full-length and four 5'-deletion fragments linked with GUS vectors were constructed and transformed into Nicotiana tabacum by Agrobacterium-mediated transformation. Transient transformation and histochemical staining of leaves indicated that the activity of the GPAT promoter was strong and induced by low-temperature stress. The deletion of a -294 bp region suggested that the DRE-motif was functionally an essential element for cold induction, and the deletion of -1494 bp and -1194 bp regions suggested that negative regulation exists in the promoter. Our results show that the GPAT gene promoter is a key regulator under cold stress and we think that this study will have significant impact on lily molecular breeding and improving the resistance of plants.

Keywords

GPAT, promoter, chromosome walking, cis-elements, 5'-deletion fragments, histochemical staining

First Page

552

Last Page

562

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