Agrobacterium rhizogenes mediated transformation combined with a visual selection for green fluorescent protein (GFP) has been applied effectively in carrot (Daucus carota L.) transformation. Carrot root discs were inoculated with A4, A4T, LBA1334 and LBA9402 strains, all bearing gfp gene in pBIN-m-gfp5-ER. The results indicate that transformed adventitious roots can be visually selected solely based on GFP fluorescence with a very high accuracy. The method requires no selection agents like antibiotics or herbicides and enables a reduction of labour and time necessary for tissue culture. Moreover, individual transformants can be easily excised from the host tissue and cultured separately. All of the 12 used carrot cultivars produced transformed adventitious roots and the frequency of discs producing GFP expressing adventitious roots varied from 13 to 85%. The highest transformation rate was found for A4T and LBA1334 strains possessing chromosomal background of A. tumefaciens C58. The results encourage that visual selection of transformed, fluorescing adventitious roots can be highly effective and applied routinely for the production of carrot transgenic plants.

Mesophyll protoplasts of Brassica oleracea var. botrytis were successfully transformed using polyethylene glycol (PEG). The success of plant transformation depended on both gene transfer and plant regeneration. Parameters, such as PEG and vector concentrations and heat shock conditions were tested in experiments on transient expression of the beta-glucuronidase (EC 3.2.1.31) gene and the most suitable conditions for DNA uptake were determined. Two antibiotic resistance marker genes for neomycin phosphotransferase (EC 2.7.1.95) and hygromycin phosphotransferase (EC 2.7.1.104), and three vector plasmids with different lengths were used to obtain stable transformants.

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Somatic hybrids were produced by PEGinduced symmetric and asymmetric protoplast fusions in order to transfer resistance to Alternaria brassicicola, A. brassicae, Phoma lingam, Plasmodiophora brassicae and Turnip mosaic virus (TuMV) into Brassica oleracea var. capitata (cv. 'Toskama') and botrytis (cv. 'Korso'). As resistance donors, ten species belonging to several genera of the family Brassicaceae including wild relatives were used. Of 2,189 plants (somatic hybrids, partially in vitro cloned) tested, 1,616 (73.8%) were resistant against at least one of the pathogens, indicating that, mostly, a successful resistance transfer has taken place. Five hundred and twenty-two hybrids showed multiple resistances to two, three and, in a single case, to four pathogens. Irrespective of the donor parents used in the fusion, a broad variability in symptom manifestation ranging from 0 (without symptoms) to 9 (highly susceptible) could be observed. With regard to the Alternaria pathogens, Sinapis alba, B. nigra and B. juncea were the most effective resistance donors, whereas fusions with Raphanus sativus resulted in the most hybrids with resistance to clubroot and TuMV. As could be shown especially in asymmetric fusions with S. alba, Barbarea vulgaris and Hesperis matronalis, transferred resistance to a pathogen may not correspond with resistance exhibited by the donor parent. Some combinations in which both parents were highly susceptible, e.g. R. sativus (?) B. oleracea var. capitata, yielded hybrids that exhibited strong resistance, e.g. to A. brassicicola, revealing that a new type of resistance might be occurring. With regard to the Alternaria pathogens, resistance expression was very unstable. Many hybrids into which (also variable) resistance of some donors, such as B. vulgaris, S. alba and B. carinata, was transferred became as highly susceptible as those of which the fusion parents did not show any resistance reaction (e.g. R. sativus). For reliable characterization of the resistance response, hybrids should be subjected to several resistance tests during growth period of the host, at least until flowering.