Isolation of a cotton reversibly glycosylated polypeptide (GhRGP1) promoter and its expression activity in transgenic tobacco

We, Ai-Min and Ling, C. and Liu, Jin-Yuan (2006) Isolation of a cotton reversibly glycosylated polypeptide (GhRGP1) promoter and its expression activity in transgenic tobacco. Journal of Plant Physiology, 163 (4). pp. 426-435.

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Reversibly glycosylated polypeptides (RGPs) are thought to be involved in polysaccharide metabolism. A cDNA of the cotton (Gossypium hirsutum) RGP gene, designated GhRGP1, has previously been characterized, and is preferentially expressed in fiber cells. In order to investigate its temporal and spatial control, we isolated a 624 bp fragment upstream of the GhRGP1 coding sequence using a polymerase chain reaction (PCR)-based genomic walking method, transcriptionally fused the 624 bp promoter sequence to the β-glucuronidase (GUS) gene, and analyzed the stable gene expression in tobacco (Nicotiana tabacum). In 4-week-old transgenic tobacco plants, the highest expression level was observed in roots, and the GUS activity was 1.13- and 6.65-fold higher than that in stems and leaves, respectively. In the reproductive growth stage, the GUS expression level was highest in the pistils and the GUS activity in the stigmas and styles were 17.6-fold higher than that in the ovaries. High GUS activity was also detected in the anthers. In addition, histochemical staining for GUS activity on transgenic tobacco plants further indicated a higher expression in the trichomes, seeds and vascular tissues of stems. Abiotic stress treatments on transgenic tobacco plants showed that wounding and dehydration induced GUS expression. These results demonstrated the spatial and temporal regulation of a cotton RGP promoter in a model plant, and provided an important insight into the factors that control the fiber development and stress responses of the gene.

Item Type: Article
Additional Information: We are grateful to members of the Laboratory of Molecular Biology at Tsinghua University for com- ments and discussions. This work was supported by grants from the State Key Basic Research and Development Plan of China (2004CB117303), the Hi-Tech Research and Development Program of China (2004AA222100, 2002AA212051 and 2002AA207006), and the National Natural Science Foundation of China (30270753).
Uncontrolled Keywords: Cotton; Promoter activity; Reversibly glycosylated polypeptide; Transgenic tobacco; Wound-induced expression
Author Affiliation: Laboratory of Molecular Biology and MOE Laboratory of Protein Science, Department of Biological Sciences and Biotechnology, Tsinghua University(China)
Subjects: Plant Production
Crop Improvement
Divisions: Other Crops
Depositing User: Mr Siva Shankar
Date Deposited: 06 Jan 2012 10:24
Last Modified: 06 Jan 2012 10:24
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