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Gene stacking in Phalaenopsis orchid enhances dual tolerance to pathogen attack

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Abstract

Cymbidium Mosaic Virus (CymMV) and Erwinia carotovora have been reported to cause severe damage to orchid plants. To enhance the resistance of orchids to both viral and bacterial phytopathogens, gene stacking was applied on Phalaenopsis orchid by double transformation. PLBs originally transformed with CymMV coat protein cDNA (CP) were then re-transformed with sweet pepper ferredoxin-like protein cDNA (Pflp) by Agrobacterium tumefaciens, to enable expression of dual (viral and bacterial) disease resistant traits. A non-antibiotic selection procedure in the second transformation minimized the potential rate of ‘stacking’ antibiotic genes in the orchid gene pool. Transgene integration in transgenic Phalaenopsis lines was confirmed by Southern blot analysis for both CP and pflp genes. Expression of transgenes was detected by northern blot analysis, and disease resistant assays revealed that transgenic lines exhibited enhanced resistance to CymMV and E. carotovora. This is the first report describing a transgenic Phalaenopsis orchid with dual resistance to phytopathogens.

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Abbreviations

AS:

acetosyringone

CaMV:

Cauliflower Mosaic Virus

CP :

coat protein

CymMV:

Cymbidium Mosaic Virus

pflp :

sweet pepper ferredoxin-like protein

GUS :

β-glucuronidase

Hph :

hygromycin phosphotransferase

PLB:

protocorm-like body

Hph :

hygromycin phosphotransferase

Ubi :

maize ubiqutin promoter

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Authors and Affiliations

  1. Graduate Institute of Plant Biology, National Taiwan University, 106, Taipei, Taiwan

    Yuan-Li Chan

  2. Institute of BioAgricultural Sciences, Academia Sinica, 115, Taiwan

    Yuan-Li Chan,  Sanjaya & Ming-Tsair Chan

  3. Department of Horticulture, Chinese Culture University, 111, Taipei, Taiwan

    Kuang-Hung Lin

  4. Institute of Life Science, National Kaohsiung Normal University, 802, Kaohsiung, Taiwan

    Li-Jen Liao

  5. Department of Life Sciences, National University of Kaohsiung, 811, Taiwan

    Wen-Huei Chen

Authors
  1. Yuan-Li Chan
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  2. Kuang-Hung Lin
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  3. Sanjaya
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  4. Li-Jen Liao
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  5. Wen-Huei Chen
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  6. Ming-Tsair Chan
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Correspondence to Ming-Tsair Chan.

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Chan, YL., Lin, KH., Sanjaya et al. Gene stacking in Phalaenopsis orchid enhances dual tolerance to pathogen attack. Transgenic Res 14, 279–288 (2005). https://doi.org/10.1007/s11248-005-0106-5

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  • DOI: https://doi.org/10.1007/s11248-005-0106-5

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