Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 98, Issue 2, pp 125–134

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

  • Barbara Raffeiner
  • Margrethe Serek
  • Traud Winkelmann
Original Paper

DOI: 10.1007/s11240-009-9545-7

Cite this article as:
Raffeiner, B., Serek, M. & Winkelmann, T. Plant Cell Tiss Organ Cult (2009) 98: 125. doi:10.1007/s11240-009-9545-7

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.

Keywords

Display life Flower senescence Genetic engineering Orchidaceae Regeneration Somatic embryogenesis 

Abbreviations

GUS

β-Glucuronidase

MS

Murashige & Skoog

TDZ

Thidiazuron

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Barbara Raffeiner
    • 1
  • Margrethe Serek
    • 2
  • Traud Winkelmann
    • 1
    • 3
  1. 1.Research Centre for HorticultureUniversity of Applied Sciences WeihenstephanFreisingGermany
  2. 2.Institute of Floriculture and Woody Plant Science, Floriculture SectionLeibniz Universitaet HannoverHannoverGermany
  3. 3.Institute of Floriculture and Woody Plant Science, Tree Nursery Science SectionLeibniz Universitaet HannoverHannoverGermany

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