Auxotrophic Complementation in Intergeneric Hybrid Cells Obtained by Electrical and Dextran-Induced Protoplast Fusion

摘要

The auxotrophic mutants, NX1 (Nicotiana plumbagintfoliaViviani, lacking nitrate reductase) and 8B9 (Hyoscyamus muticus, tryptophan requiring) were used in protoplast fusion experiments using both electrical stimulation and dextran as fusion agents. In both cases growing colonies were recovered when the fused protoplasts were plated directly in minimal medium lacking tryptophan with nitrate as the nitrogen source. The parental cells did not survive on this medium. A cold pretreatment for 1 h at 4°C was necessary to recover hybrids after electrical fusion. The putative hybrids continued to grow on minimal medium, contained nitrate reductase activity and had alcohol dehydrogenase and esterase isozyme patterns at least in part in common with those of the parents. Two new esterase bands were expressed in the hybrids. These studies show that both electrical and dextran induced protoplast fusion can produce hybrids selectable by auxotrophic complementation