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Recovery of virus-infected Dendrobium orchids by constitutive expression of the cymbidium mosaic virus coat protein gene

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Abstract

A common practice in the production of viral disease-resistant plant varieties is the transfer of the viral coat protein (CP) gene to a wide range of healthy plants. This is the first report on the effect of a bombarded CP transgene on the replication of cymbidium mosaic virus (CymMV) in naturally infected Dendrobium orchids. Nine months after transformation, two naturally CymMV-infected transgenic Dendrobium orchids exhibited a gradual decrease in CymMV-CP transcript and coat protein levels as detected by RT-PCR and western blot hybridization, respectively. At 18 months, the transgenic lines were completely free of endogenous CymMV contamination and were protected from CymMV re-infection following inoculation. Six weeks after inoculation with purified CymMV, these virus-free transgenic lines contained no viral coat protein transcript, replicase transcript or particle accumulation from the re-invading CymMV, as detected by RT-PCR and Immunoelectron microscopy assays, respectively. Additionally, no small RNAs derived from viral CymMV-CP were detected, but the transcribed CymMV-CP positive-strand derived from the transgene appeared in these transgenic lines.

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Acknowledgments

This research was supported by The Royal Golden Jubilee Ph.D. Program of Thailand Research Fund (PHD/0197/2549) and Kasetsart University Research and Development, Thailand (V-T/D/25.51). We are grateful to Dr. William James (Jim) Peacock and Dr. Narayana M. Upadhyaya (Plant Industry, the Commonwealth Scientific and Industrial Research Organisation, ACT, Australia) for the gift of the plasmids used in this study. We thank Dr. James M. Dubbs for a critical reading of the revised manuscript.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Genetics, Faculty of Science, Kasetsart University, Chatuchak, Bangkok, 10900, Thailand

    Udomporn Petchthai, Anchalee Chuphrom & Pattana Srifah Huehne

  2. Laboratory of Biotechnology, Chulabhorn Research Institute, Laksi, Bangkok, 10210, Thailand

    Pattana Srifah Huehne

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  1. Udomporn Petchthai
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  2. Anchalee Chuphrom
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  3. Pattana Srifah Huehne
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Correspondence to Pattana Srifah Huehne.

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Petchthai, U., Chuphrom, A. & Huehne, P.S. Recovery of virus-infected Dendrobium orchids by constitutive expression of the cymbidium mosaic virus coat protein gene. Plant Cell Tiss Organ Cult 120, 597–606 (2015). https://doi.org/10.1007/s11240-014-0626-x

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