Articles
TRANSFORMATION OF ROSES WITH GENES FOR ANTIFUNGAL PROTEINS
Article number
547_3
Pages
27 – 33
Language
Abstract
In Northern Europe the cultivation of garden roses is tremendously impaired by fungal diseases.
In order to obtain partial resistance to the major diseases blackspot, powdery mildew, downy mildew and rust simultaneously, we followed the biotechnological approach to overexpress genes for different antifungal proteins.
First a protocol for transformation by agrobacterium mediated gene transfer and regeneration via somatic embryogenesis was established.
The transformation frequency was up to 3 %. According to this protocol different combinations of antifungal resistance genes, encoding a Class II chitinase, a Class II ß-1,3-glucanase and a Type I ribosome inhibiting protein from barley as well as the antibacterial T4-lysozyme gene were introduced into the garden rose cultivars Heckenzauber and Pariser Charme.
About 80 true transgenic plants were analysed for expression of their transgenes, somaclonal variation, and resistance to blackspot.
Overall, in 80 % of the putative transgenic plants expression of the transgenes could be detected by Northern analysis.
Compared to non transgenic control plants, the susceptibility to blackspot did not decrease in the case of cytosolic expression of the antifungal proteins.
The secretion of the ribosome inhibiting protein into the extracellular space, however, reduced the susceptibility against blackspot to 60 % in the mean compared to non transgenic control plants.
Several transgenic plants showed severe morphological deviations, mainly in leaf shape and flower morphology, which may be the result of both, somaclonal variation and the transformation procedure.
In order to obtain partial resistance to the major diseases blackspot, powdery mildew, downy mildew and rust simultaneously, we followed the biotechnological approach to overexpress genes for different antifungal proteins.
First a protocol for transformation by agrobacterium mediated gene transfer and regeneration via somatic embryogenesis was established.
The transformation frequency was up to 3 %. According to this protocol different combinations of antifungal resistance genes, encoding a Class II chitinase, a Class II ß-1,3-glucanase and a Type I ribosome inhibiting protein from barley as well as the antibacterial T4-lysozyme gene were introduced into the garden rose cultivars Heckenzauber and Pariser Charme.
About 80 true transgenic plants were analysed for expression of their transgenes, somaclonal variation, and resistance to blackspot.
Overall, in 80 % of the putative transgenic plants expression of the transgenes could be detected by Northern analysis.
Compared to non transgenic control plants, the susceptibility to blackspot did not decrease in the case of cytosolic expression of the antifungal proteins.
The secretion of the ribosome inhibiting protein into the extracellular space, however, reduced the susceptibility against blackspot to 60 % in the mean compared to non transgenic control plants.
Several transgenic plants showed severe morphological deviations, mainly in leaf shape and flower morphology, which may be the result of both, somaclonal variation and the transformation procedure.
Authors
A. Dohm, C. Ludwig, D. Schilling, T. Debener
Keywords
Agrobacterium mediated gene transfer, somatic embryogenesis, embryogenic callus, ribosome inhibiting protein, chitinase, glucanase, T4-lysozyme, signal peptide, fungal disease resistance
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