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Generating of rice OsCENH3-GFP transgenic plants and their genetic applications

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Chinese Science Bulletin

Abstract

In order to investigate rice functional centromeres, OsCENH3-GFP chimeric gene was constructed and transformed into the indica rice variety, Zhongxian 3037, mediated by Agrobacturium. The integration of the exogenous genes in the transgenic plants was confirmed by PCR and Southern blotting. The transgenic plants grow normally during their whole life time, just like Zhongxian 3037. No significant defects were detected in either mitosis or meiosis of the transgenic plants. The overlapping of GFP signals and anti-CENH3 foci in both mitotic and meiotic cells from T0 and T1 generation plants indicated that GFP had been successfully fused with CENH3, so the GFP signals can well represent the CENH3 locations on each chromosome. To evaluate the applicability of the transgenic plants to other genetic studies, fluorescence in situ hybridization (FISH) using rice centromeric tandem repetitive sequence CentO as the probe was conducted on the zygotene chromosomes of pollen mother cells (PMCs). It has been revealed that the GFP signals are overlapping with CentO FISH signals, showing that CentO is one of the key elements constituting rice functional centromeres. Immunofluorescent staining using anti-α-tublin antibody and anti-PAIR2 antibody on the chromosomes during mitosis and meiosis stages of the transgenic plants further reveals that OsCENH3-GFP transgenic plants can be widely used for studying rice molecular biology, especially for tagging functional centromeres in both living cells and tissues.

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Authors and Affiliations

  1. Key Laboratory of Crop Genetics and Physiology of Jiangsu Province/Key Laboratory of Plant Functional Genomics of Ministry of Education, Yangzhou University, Yangzhou, 225009, China

    HengXiu Yu, Xin Wang, ZhiYun Gong & MingHong Gu

  2. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China

    Ding Tang & ZhuKuan Cheng

Authors
  1. HengXiu Yu
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  2. Xin Wang
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  3. ZhiYun Gong
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  4. Ding Tang
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  5. MingHong Gu
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  6. ZhuKuan Cheng
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Corresponding author

Correspondence to ZhuKuan Cheng.

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Contributed equally to this work

Supported by the National Natural Science Foundation of China (Grants Nos. 30770131, 30530070 and 30671285) and High Technology Research and Development Program of China (Grant No. 2006AA02Z124)

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Yu, H., Wang, X., Gong, Z. et al. Generating of rice OsCENH3-GFP transgenic plants and their genetic applications. Chin. Sci. Bull. 53, 2981–2988 (2008). https://doi.org/10.1007/s11434-008-0376-4

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  • DOI: https://doi.org/10.1007/s11434-008-0376-4

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