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THE USE OF PROTOPLAST FUSION AND TRANSFORMATION FOR THE PRODUCTION OF ASYMMETRIC SOMATIC HYBRIDS

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In asymmetric hybridization experiments, a Nicotiana paniculata cell line transformed with the Agrobacterium tumefaciens strain B6S3 was used as the donor plant material. This cell line exhibits two dominant marker genes, hormone-independent growth behavior and synthesis of octopine, as a result of the incorporation of T-DNA from Agrobacterium. The recipient cell line was the cnx-68 N. tabacum cell line which has two recessive markers: nitrate reductase deficiency and presence of an intracellular calcium-oxalate druse. Isolated protoplasts of the donor cell line N. paniculata B6S3 were mitotically inactivated by X-rays and fused with protoplasts of the cell line cnx-68. Asymmetric somatic hybrids were selected on hormone-free agar medium supplemented with 50 mM KClO3. Eighteen cell lines survived the selection treatment; seven of them exhibited the two dominant and the two recessive markers. In a second experiment, the marker gene hygromycin resistance was introduced into Brassica nigra by using a disarmed Agrobacterium strain. The B. nigra lines employed possessed resistance against two important rape pathogens (Phoma lingam and Plasmodiophora brassicae). Asymmetric somatic hybrids were obtained by fusion of X-ray irradiated B. nigra protoplasts with protoplasts of B. napus, and hygromycin was used for their selection. The obtained asymmetric somatic hybrids contained variable amounts of the B. nigra genome. The possibility of a cotransfer of the two disease resistances will be studied.

Affiliations: 1: Institut für Angewandte Genetik, Freie Universität Berlin, Albrecht-Thaer-Weg 6 D-1000 Berlin 33, F.R. of Germany ; 2: Institut für Angewandte Genetik, Freie Universität Berlin

10.1080/0021213X.1991.10677182
/content/journals/10.1080/0021213x.1991.10677182
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/content/journals/10.1080/0021213x.1991.10677182
1991-05-13
2018-09-19

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