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Isolation and characterization of Brittle2 promoter from Zea Mays and its comparison with Ze19 promoter in transgenic tobacco plants

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Abstract

ADP-glucose pyrophosphorylase (AGPase) represents a key regulatory step in starch synthesis. A 0.9 kb of 5′ flanking region preceding Brittle2 gene, encoding the small subunit of maize endosperm AGPase, was cloned from maize genome and its expression pattern was studied via the expression of β-glucuronidase (GUS) gene in transgenic tobacco. Analysis of GUS activities showed that the 0.9 kb fragment flanking Brittle2 gene was sufficient for driving the seed-preferred expression of the reporter gene. The activity of the 0.9 kb 5′ flanking fragment was compared with that of the tandem promoter region from a zein gene (zE19, encoding a maize 19 kDa zein protein). The results indicated that both promoters were seed-preferred in a dicotyledonous system as tobacco and the activity of zE19 promoter was three to fourfold higher than that of the 0.9 kb fragment flanking Brittle2 gene in transgenic tobacco seeds. At the same time, zE19-driven GUS gene expressed earlier than Brittle2 promoter during seed development. Histochemical location of GUS activity indicated that both promoters showed high expression in embryos, which is different from similar promoters tested in maize.

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Abbreviations

CaMV:

Cauliflower mosaic virus

DAF:

Days after flowering

4-MU:

4-methylumbelliferone

GUS:

β-glucuronidase

MUG:

4-methylumbelliferyl-β-d-glucuronide

X-gluc:

5-bromo-4-chloro-3-indolyl-β-d-glucuronide

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Acknowledgements

We thank Prof. Gong Zhizhong (China Agricultural University) for use of anatomical lens. This work was supported by the National Special Program for Research and Industrialization of Transgenic Plants and the Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China (No: 705009).

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Authors and Affiliations

  1. State Key Laboratory of Agrobiotechnology and National Center for Maize Improvement, China Agricultural University, Beijing, 100094, China

    Xiaoping Chen, Zhangying Wang, Jianhua Wang, Maoyan Wang, Li Zhao & Guoying Wang

  2. Institute of Crop Sciences, CAAS, Beijing, 100081, China

    Guoying Wang

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  1. Xiaoping Chen
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  2. Zhangying Wang
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  3. Jianhua Wang
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  4. Maoyan Wang
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  5. Li Zhao
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  6. Guoying Wang
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Correspondence to Guoying Wang.

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Chen, X., Wang, Z., Wang, J. et al. Isolation and characterization of Brittle2 promoter from Zea Mays and its comparison with Ze19 promoter in transgenic tobacco plants. Plant Cell Tiss Organ Cult 88, 11–20 (2007). https://doi.org/10.1007/s11240-006-9165-4

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  • DOI: https://doi.org/10.1007/s11240-006-9165-4

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