ISHS Acta Horticulturae 447: III International Symposium on In Vitro Culture and Horticultural Breeding
GENETIC MANIPULATION OF HAPLOID DATURA INNOXIA MILL.: ANALYSIS OF THE TRANSGENE INTEGRATION PATTERNS AND THE PLOIDY LEVEL OF TRANSGENIC PLANTS OBTAINED AFTER DIRECT OR AGROBACTERIUM-MEDIATED GENE TRANSFER |
| Authors: | M. Meixner, C. Frahm, P. Pflug, T. Schmidt-Rogge, O. Schieder |
| Abstract:
We have transformed haploids of the solanaceous species Datura innoxia using either direct (PEG-mediated DNA transfer into protoplasts) or Agrobacterium-mediated gene transfer (cocultivation of different explants together with binary strains). We found that many of regenerated transgenics turned into diploid plants. A significant increase in the percentage of transformed haploids was observed when using the Agrobacterium-approach (30%) compared to the number of haploid plants obtained after PEG-mediated gene transfer (10%). Plants obtained via Agrobacterium-mediated gene transfer normally are harboring one to three copies of the T-DNA and approximately 50% have a single T-DNA integration in their genome. Plants regenerated from protoplast born callus, after direct gene transfer, were found to contain several copies of the vector DNA. Only a few plants with a single copy integration were recovered. In contrast, for haploid Datura innoxia the protoplast system is very easy to handle and regeneration is faster when compared to the Agrobacterium-based system. | |
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