Transformation of different barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) cultivars by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> infection of in vitro cultured ovules |
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Authors: | Inger Bæksted Holme Henrik Brinch-Pedersen Mette Lange Preben Bach Holm |
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Institution: | (1) Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, Research Centre Flakkebjerg, University of Aarhus, 4200 Slagelse, Denmark |
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Abstract: | Most cultivars of higher plants display poor regeneration capacity of explants due to yet unknown genotypic determined mechanisms.
This implies that technologies such as transformation often are restricted to model cultivars with good tissue characteristics.
In the present paper, we add further evidence to our previous hypothesis that regeneration from young barley embryos derived
from in vitro-cultured ovules is genotype independent. We investigated the ovule culture ability of four cultivars Femina,
Salome, Corniche and Alexis, known to have poor response in other types of tissue culture, and compared that to the data for
the model cultivar, Golden Promise. Subsequently, we analyzed the transformation efficiencies of the four cultivars using
the protocol for Agrobacterium infection of ovules, previously developed for Golden Promise. Agrobacterium tumefaciens strain AGL0, carrying the binary vector pVec8-GFP harboring a hygromycin resistance gene and the green fluorescence protein
(GFP) gene, was used for transformation. The results strongly indicate that the tissue culture response level in ovule culture
is genotype independent. However, we did observe differences between cultivars with respect to frequencies of GFP-expressing
embryos and frequencies of regeneration from the GFP-expressing embryos under hygromycin selection. The final frequencies
of transformed plants per ovule were lower for the four cultivars than that for Golden Promise but the differences were not
statistically significant. We conclude that ovule culture transformation can be used successfully to transform cultivars other
than Golden Promise. Similar to that observed for Golden Promise, the ovule culture technique allows for the rapid and direct
generation of high quality transgenic plants. |
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Keywords: | Agrobacterium tumefaciens Barley cultivars Ovule culture Transformation Zygote |
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