Auxotroph Complementation via Protoplast Fusion in Hyoscyamus Muticus and Nicotiana Tabacum

Summary

Routine plant regeneration from haploid protoplasts of Hyoscyamus mutious (egyptian henbane) has been established as basis for auxotroph selection (Lörz et al., 1979; Wernicke et al., 1979). A series of four histidine-, one tryptophan-, three nicotinamide-, and one leucine-requiring auxotrophs of four nitrate nonutilizing lines and of two temperature-sensitive clones have been recovered (Strauss et al., 1981; Gebhardt et al., 1981). Protoplast fusion and subsequent selection by complementation led to the recovery of wild-type clones in various combinations, e.g., his^- + trp^-, nic^- + nic^-, nic^- + NR^-, ts + ts, ts + trp^-, ts + his^-. Intergeneric combination of one of the NR^- auxotrophs of Eyosoyamus mutious with both the defined NR^- mutants nia-115 and cnx^-68 of Nicotiana tabacum (Müller and Grafe, 1978) yielded wild-type clones with the apoenzyme mutant exclusively. Intergeneric combination of one of the nic^- clones of Eyosoyamus and the NR mutant cnx-68 of Niootiana gave wild-type clones with high morphogenetic potential producing a variety of different morphological types including flowering and fertile plants. Crossing analysis and anther culture analysis of these plants is in progress.