Agrobacterium tumefaciens-mediated transformation of Oncidium and Odontoglossum orchid species with the ethylene receptor mutant gene etr1-1 |
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Authors: | Barbara Raffeiner Margrethe Serek and Traud Winkelmann |
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Institution: | (1) Research Centre for Horticulture, University of Applied Sciences Weihenstephan, Am Staudengarten 8, 85354 Freising, Germany;(2) Institute of Floriculture and Woody Plant Science, Floriculture Section, Leibniz Universitaet Hannover, Herrenhaeuser Str. 2, 30419 Hannover, Germany;(3) Institute of Floriculture and Woody Plant Science, Tree Nursery Science Section, Leibniz Universitaet Hannover, Herrenhaeuser Str. 2, 30419 Hannover, Germany |
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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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Keywords: | Display life Flower senescence Genetic engineering Orchidaceae Regeneration Somatic embryogenesis |
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