Abstract
Protoplasts of the nicotinamide-deficient Hyoscyamus muticus cell line nic− IVH2 and of the nitrate reductase-deficient Nicotiana tabacum cell line NR− cnx 68 were induced to fuse. Selection for putative interspecific hybrid clones was via auxotroph complementation. Controls included tests for cross-feeding, cross inhibition, PEG-induced variation, culture-induced variation, reversion, viability, delayed selection. Protrotrophic cell lines were recovered exclusively from PEG-treated mixtures of both protoplast types. The putative hybrid clones grew independent of exogenously supplied auxins and cytokinins, and at a faster rate than either parent. The morphogenic potential of different clones varied from ‘non-morphogenic’ to ‘regeneration of fertile plants’. Indirect evidence for the hybrid nature of the clones is provided from a) tight selection, b) hormone-independent growth, c) hybrid vigour, d) extreme morphological variation, e) isoenzyme bands from both parents, f) morphogenic potential. Definite proof for the hybrid nature was, however, provided from species-specific DNA hybridization. Chimerism could be excluded since only the large subunit of Hyoscyamus muticus ribulose bisphosphate carboxylase was found and since species-specific DNA hybridization identified clones which gave no Nicotiana tabacum signal.
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Abbreviations
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- NAA:
-
Naphthaleneacetic acid
- BAP:
-
6-Benzylaminopurine
- PEG:
-
Polyethyleneglykol
- MES:
-
(2[N-morpholino]ethane sulfonic acid
- Tris-HCl:
-
Tris(hydroxymethyl)-aminoethane-HCl % is given throughout in w/v
- FMI:
-
Friedrich Miescher-Institut
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Communicated by K. Hahlbrock
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Potrykus, I., Jia, J., Lazar, G.B. et al. Hyoscyamus muticus + Nicotiana tabacum fusion hybrids selected via auxotroph complementation. Plant Cell Reports 3, 68–71 (1984). https://doi.org/10.1007/BF00270974
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DOI: https://doi.org/10.1007/BF00270974