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Agrobacterium tumefaciens-mediated transformation of Oncidium and Odontoglossum orchid species with the ethylene receptor mutant gene etr1-1

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Abstract

Oncidium and Odontoglosum orchid species have reduced display lives and are thus commercially less important than Phalaenopsis. One approach to prolonging display life permanently is to transform Oncidium and Odontoglossum with the ethylene receptor mutant gene etr1-1 from Arabidopsis under control of a flower specific promoter; this should reduce their sensitivity to exogenous ethylene. To achieve this it will be necessary to establish an efficient regeneration protocol using somatic embryogenesis and a routine Agrobacterium tumefaciens-mediated transformation procedure. Protocorm-like bodies (PLBs) of both orchid genera were regenerated from leaf tip explants. Leaf tips and PLBs, cultured in liquid and solid media, were compared as targets for genetic transformation. No transgenic shoots were obtained from leaf tips, while PLBs of Oncidium and Odontoglossum cultured on solid medium were successfully transformed with an expression vector containing nptII and gus genes driven by the cauliflower mosaic virus (CaMV) 35S promoter. Applying the A. tumefaciens strain EHA 105, transformation efficiencies of 1.3–2.7% were achieved for the investigated genotypes. Transformation with etr1-1 gene was achieved subsequently. Oncidium ‘Sweet Sugar’ has been successfully transformed and validated by PCR and Southern analysis.

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Abbreviations

GUS:

β-Glucuronidase

MS:

Murashige & Skoog

TDZ:

Thidiazuron

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Acknowledgments

This research was supported by the grant for PhD study (BR) from Valtl Raffeiner, Gartenbau Valtl Raffeiner, Italy. The authors thank Dr. Viola Mußmann and Dr. Heiko Mibus-Schoppe for their help with Southern hybridisations, and Prof. Errol Hewett (Massey University, Palmerston North, New Zealand) for linguistic editing of the manuscript.

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Authors and Affiliations

  1. Research Centre for Horticulture, University of Applied Sciences Weihenstephan, Am Staudengarten 8, 85354, Freising, Germany

    Barbara Raffeiner & Traud Winkelmann

  2. Institute of Floriculture and Woody Plant Science, Floriculture Section, Leibniz Universitaet Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany

    Margrethe Serek

  3. Institute of Floriculture and Woody Plant Science, Tree Nursery Science Section, Leibniz Universitaet Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany

    Traud Winkelmann

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  1. Barbara Raffeiner
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Correspondence to Traud Winkelmann.

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Raffeiner, B., Serek, M. & Winkelmann, T. Agrobacterium tumefaciens-mediated transformation of Oncidium and Odontoglossum orchid species with the ethylene receptor mutant gene etr1-1 . Plant Cell Tiss Organ Cult 98, 125–134 (2009). https://doi.org/10.1007/s11240-009-9545-7

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