An efficient and largely genotype-independent transformation method for Brassica napus and Brassica oleracea was established based on neo or bar as selectable marker genes. Hypocotyl explants of Brassica napus and Brassica oleracea cultivars were infected with Agrobacterium strains containing chimeric neo and bar genes. The use of AgNO3 was a prerequisite for efficient shoot regeneration under selective conditions. Vitrification was avoided by decreasing the water potential of the medium, by decreasing the relative humidity in the tissue culture vessel, and by lowering the cytokinin concentration. In this way, rooted transformed shoots were obtained with a 30% efficiency in 9 to 12 weeks. Southern blottings and genetic analysis of SI-progeny showed that the transformants contained on average between one and three copies of the chimeric genes. A wide range of expression levels of the chimeric genes was observed among independent transformants. Up to 25% of the transformants showed no detectable phosphinotricin acetyltransferase or neomycin phosphotransferase 11 enzyme activities although Southern blottings demonstrated that these plants were indeed transformed.
Rhizomania is a disease of sugar beet caused by the furovirus beet necrotic yellow vein virus (BNYVV) . Coat protein mediated resistance has been reported for a number of viral diseases . This approach to virus resistance was therefore attempted for control of rhizomania . Two constructs of the coat protein gene of BNYVV were introduced into sugar beet by Agrobacterium-mediated transformation. The mRNA level was estimated to be 0 .01 % of the poly A+ RNA . Expression of the coat protein gene was under the detection limit of our western blotting protocol i .e . below 0 .01 µg /50 pg (0 .02% of the total soluble protein) . One transformation event per construct was tested in a greenhouse assay and in rhizomania infested soil in a field trial . In the greenhouse assay, transgenic plants showed a strong reduction of virus multiplication when compared to non-transgenic plants . This result was confirmed in the field trial, where a significant difference in virus multiplication was shown between plants with and without the coat protein gene .
Antifungal rhizobacteria were obtained from maize, barley and chicory using direct or indirect isolation procedures. Effective isolates were tested for broad-spectrum activity against a set of phytopathogenic fungi. Isolates with broad-spectrum activity were identified as Pseudomonas fluorescens, P. cepacia, Serratia liquefaciens, S. plymuthica and Bacillus sp. Broad-spectrum compounds produced by P. cepacia and Erwinia herbicola were characterized as pyrrolnitrin and herbicolin-like compounds respectively
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