Transformation of different barley (Hordeum vulgare L.) cultivars by Agrobacterium tumefaciens infection of in vitro cultured ovules

• Published in: Plant cell reports 2008-12, Vol.27 (12), p.1833-1840
• Main Authors: Holme, Inger Bæksted, Brinch-Pedersen, Henrik, Lange, Mette, Holm, Preben Bach
• Format: Article
• Language: English
• Subjects: Agrobacterium tumefaciens; Agrobacterium tumefaciens - genetics; Barley; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Cell Biology; Cultivars; Culture techniques; Embryos; Flowers - microbiology; Fundamental and applied biological sciences. Psychology; Gene Dosage; Gene Expression; Genetic Transformation and Hybridization; Genetic Vectors; Green Fluorescent Proteins - genetics; Hordeum - embryology; Hordeum - genetics; Hordeum - microbiology; Hordeum vulgare; Life Sciences; Plant Biochemistry; Plant Sciences; Plants, Genetically Modified; Plasmids - genetics; Recombinant Proteins - genetics; Tissue Culture Techniques; Transformation, Genetic; Transgenic plants
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-008-0605-y

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.