Original Article
Cell Research (2007) 17: 174–183. doi: 10.1038/sj.cr.7310119; published online 19 December 2006
Functional analysis of a cotton glucuronosyltransferase promoter in transgenic tobaccos
Ai-Min Wu1, Shi-You Lv1 and Jin-Yuan Liu1
1Laboratory of Molecular Biology and Protein Science Laboratory of the Ministry of Education, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China
Correspondence: Jin-Yuan Liu, Tel: +86-10-62772243; E-mail: liujy@mail.tsinghua.edu.cn
Received 3 August 2006; Revised 25 October 2006; Re-revised 8 November 2006; Accepted 28 November 2006.
Abstract
The 5' fragment (1 647 bp) of the cotton glucuronosyltransferase gene (GhGlcAT1) was transcriptionally fused to the
-glucuronidase (GUS) gene, and functionally analyzed for important regulatory regions controlling gene expression in transgenic tobacco plants. GUS activity analysis revealed that the full-length promoter drives efficient expression of the GUS gene in the root cap, seed coat, pollen grains and trichomes. Exposure of the transgenic tobacco to various abiotic stresses showed that the promoter was mainly responsive to the sugars (glucose and sucrose) as well as gibberellic acid. Progressive upstream deletion analyses of the promoter showed that the region from -281 to +30 bp is sufficient to drive strong GUS expression in the trichomes of shoot, suggesting that the 311 bp region contains all cis-elements needed for trichome-specific expression. Furthermore, deletion analysis also revealed that the essential cis-element(s) for sucrose induction might be located between -635 and -281 bp. In addition, sequence analysis of the regulatory region indicated several conserved motifs among which some were shared with previously reported seed-specific elements and sugar-responsive elements, while others were related with trichome expression. These findings indicate that a 1 647-bp fragment of the cotton GhGlcAT1 promoter contains specific transcription regulatory elements, and provide clues about the roles of GhGlcAT1 in cotton fiber development. Further analyses of these elements will help to elucidate the molecular mechanisms regulating the expression of the GhGlcAT1 gene during fiber elongation.
Keywords:
glucuronosyltransferase, cotton, sugar-induced expression, regulatory elements, promoter activity, transgenic tobacco
Abbreviations:
GlcAT, glucuronosyltransferase; DAF, days after flowering; 4-MU, 4-methylumbelliferone; Suc, sucrose; Glu, glucose; Fru, fructose; Man, mannitol; Sor, sorbitol; Eth, ethylene
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