Two novel positive cis-regulatory elements involved in green tissue-specific promoter activity in rice (Oryza sativa L ssp.)

Ye , R. and Zhou, F. and Lin , Y. (2012) Two novel positive cis-regulatory elements involved in green tissue-specific promoter activity in rice (Oryza sativa L ssp.). Plant Cell Reports, 31 (7). pp. 1159-1172.

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In plant genetic engineering, using tissue-specific promoters to control the expression of target gene is an effective way to avoid potential negative effects of using constitutive promoter, such as metabolic burden and so on. However, until now, there are few tissue-specific promoters with strong and reliable expression that could be used in crop biotechnology application. In this study, based on microarray and RT-PCR data, we identified a rice green tissue-specific expression gene DX1 (LOC_Os12 g33120). The expression pattern of DX1 gene promoter was examined by using the β-glucuronidase (GUS) reporter gene and analyzed in transgenic rice plants in different tissues. Histochemical assays and quantitative analyses of GUS activity confirmed that PDX1:GUS was highly expressed in green tissues. To identify the regulatory elements controlling the expression of the DX1 gene, a series of 5′ and 3′ deletions of DX1 promoter were fused to GUS gene and stably introduced into rice plants. In addition, gel mobility shift assays and site-directed mutagenesis studies were used, allowing for the identification of two novel tissue-specific cis-acting elements (GSE1 and GSE2) within PDX1. GSE1 acted as a positive regulator in all green tissues (leaf, sheath, stem and panicle). Compared with GSE1, GSE2 acted as a positive regulator only in sheath and stem tissue, and had a weaker effect on gene expression. In addition, PDX1:GUS was not expressed in anther and seed, this characteristic reduced the potential ecological risk and potential food safety issues. Taken together, our results strongly suggest that the identified promoter, PDX1, and its cis regulatory elements, GSE1 and GSE2, are potentially useful in the field of rice transgenic breeding.

Item Type: Article
Additional Information: This research was funded by the National Program of Transgenic Variety Development of China (2011ZX11001- 001, the development of insect-resistant transgenic rice).
Uncontrolled Keywords: Promoter, Cis-element, Green tissue-specific, GUS gene, Rice
Author Affiliation: National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research, Huazhong Agricultural University, China.
Subjects: Crop Improvement
Crop Improvement > Genetics/Genomics
Crop Improvement > Biotechnology
Divisions: Other Crops
Depositing User: Mr Arbind Seth
Date Deposited: 12 Dec 2012 12:06
Last Modified: 12 Dec 2012 12:06
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