Asymmetric hybridization in Nicotiana by fusion of irradiated protoplasts

Bates, G.W.; Hasenkampf, C.A.; Contolini, C.L.; Piastuch, W.C.

Tag. Theoretical and Applied Genetics. Theoretische und Angewandte Genetik 74(6): 718-726


ISSN/ISBN: 0040-5752
PMID: 24240331
DOI: 10.1007/bf00247548
Accession: 001533488

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Mesophyll protoplasts of a kanamycin-resistant, nopaline-positive N. plumbaginifolia seed line were inactivated by gamma -irradiation and electrically fused with unirradiated mesophyll protoplasts of N. tabacum. Hybrids were selected on kanamycin. All of the regenerants resembled N. tabacum in morphology and many were indistinguishable from it. In 37 plants, one or two esterases characteristic of N. plumbaginifolia were found in addition to a full set of esterases from N. tabacum. These plants were classified as somatic hybrids, but irradiation had clearly reduced the amount of N. plumbaginifolia genetic material they contained. Twenty-four plants were found that had only N. tabacum esterases but that produced nopaline and were kanamycin resistant; Southern blotting revealed the presence of the kanamycin resistance gene. They were designated asymmetric hybrids. Finally, 25 plants were regenerated that were sensitive to kanamycin, did not synthesize nopaline and contained only N. tabacum esterases. All the regenerated plants, including this last category, were male sterile. Most of the asymmetric hybrids contained one more chromosome than is normal for N. tabacum. They were successfully backcrossed as females to N. tabacum, but the kanamycin-resistance gene was partially unstable during meiosis, as would be expected for factors inherited on an impaired chromosome.

Asymmetric hybridization in Nicotiana by fusion of irradiated protoplasts