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RNA Silencing Mediated by Direct Repeats in Maize: A Potential Tool for Functional Genomics

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Abstract

It has been shown in tobacco and Arabidopsis that transgenes with multiple direct repeats induce RNA silencing at high frequency. In this study, we tried to establish a direct repeat-induced RNA silencing system in maize and evaluate whether it can be developed as a high throughput tool for functional genomics. Our results showed that the construct phC4, which carries four direct repeats of a chloramphenicol acetyl-transferase (CAT) gene, was able to induce silencing of itself with high efficiency in maize. Using a transient expression system, we further demonstrated that construct phC3G with a β-glucuronidase (GUS) gene located downstream of three direct repeats of CAT gene silenced not only itself in maize calli but also an “endogenous” GUS gene, which was stably expressed in maize calli. Most importantly, when constructs with the maize iojap (ij) gene inserted in either sense or antisense orientation into the downstream of four direct repeats of CAT gene were transformed into maize plants, co-suppression of endogenous and transgenic ij genes was detected in majority of transgenic maize plants. Our co-suppression results suggest that with improvements, this new approach has the potential to become an efficient research tool for high throughput functional genomics.

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Acknowledgments

We would like to thank Prof. Qifeng Xu, China Agricultural University, for providing maize inbred lines; Dr. Amitava Mitra, University of Nebraska-Lincoln, for providing the pC4 and pC3G constructs; Dr. Chengxia Li and Dr. Xiangbai Dong, University of California and the Chinese Academy of Sciences, respectively, for critical reading of this manuscript. This research was supported by program for “New Century Excellent Talents in University, NCET-05-0129”.

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  1. Xiuping Xu

    Present address: Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, The Chinese Academy of Sciences, Beijing, 100093, China

Authors and Affiliations

  1. State Key Laboratory for Agro-biotechnology, College of Biological Sciences, China Agricultural University, No. 2 Yuanmingyuan West Road, Haidian District, Beijing, 100094, People’s Republic of China

    Xiuping Xu, Dengyun Zhu, Qian Zhao, Guangming Ao & Jingjuan Yu

  2. Department of Entomology, University of Arizona, Tucson, AZ, 85721-0036, USA

    Chonglie Ma

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  1. Xiuping Xu
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Correspondence to Chonglie Ma or Jingjuan Yu.

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Xu, X., Zhu, D., Zhao, Q. et al. RNA Silencing Mediated by Direct Repeats in Maize: A Potential Tool for Functional Genomics. Mol Biotechnol 41, 213–223 (2009). https://doi.org/10.1007/s12033-008-9124-x

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