Combination of kanamycin resistance and nitrate reductase deficiency as selectable markers in one nuclear genome provides a universal somatic hybridizer in plants

Brunold, Christian; Krüger-Lebus, Susanne; Saul, Michael W.; Wegmüller, Samuel; Potrykus, Igo (1987). Combination of kanamycin resistance and nitrate reductase deficiency as selectable markers in one nuclear genome provides a universal somatic hybridizer in plants. Molecular and general genetics MGG, 208(3), pp. 469-473. Springer 10.1007/BF00328141

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The combination in the nuclear genome of a dominant resistance marker (to select against unfused wild-type cells) and a recessive deficiency marker (to select against unfused mutant cells) in a cell line should provide a system for selecting fusion hybrids between the mutant line and any wild-type line. To test this idea, we fused protoplasts from a non-morphogenic cell line of Nicotiana tabacum which was kanamycin resistant (by transformation) and deficient in nitrate reductase (NR-K+) with protoplasts from N. tabacum cv. Petit Havana clone SR1, which provided resistance against streptomycin as an additional selectable marker (NR+K-SR+). Putative hybrids were selected using a culture medium containing no available reduced nitrogen source and 50 mg/l kanamycin sulphate. After regeneration into plants, the hybrid character was demonstrated from: (i) the morphological variation of the regenerants; (ii) the chromosome number; (iii) the ability to grow on medium without a reduced nitrogen source and containing kanamycin sulphate at 50 mg/l; (iv) the presence of nitrate reductase activity; (v) the presence of the gene coding for neomycin phosphotransferase, which provides resistance to kanamycin sulphate; (vi) callus formation from leaves on medium containing 1 g/l streptomycin or 50 mg/l kanamycin sulphate; (vii) F1 plants containing nitrate reductase and the gene for neomycin phosphotransferase. Fusions between the mutant cell line (NR-K+) and three wild-type tobacco species and subsequent cultivation on medium containing no available nitrogen source but 50 mg/l kanamycin sulphate resulted in callus formation with all combinations, while hybrid plants were only regenerated when N. sylvestris was the fusion partner.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Department of Biology > Institute of Plant Sciences (IPS) > Stress Physiology (discontinued)
08 Faculty of Science > Department of Biology > Institute of Plant Sciences (IPS)

UniBE Contributor:

Brunold, Christian and Wegmüller, Samuel

Subjects:

500 Science > 580 Plants (Botany)

ISSN:

0026-8925

Publisher:

Springer

Language:

English

Submitter:

Peter Alfred von Ballmoos-Haas

Date Deposited:

16 Jun 2017 13:46

Last Modified:

14 Nov 2017 16:31

Publisher DOI:

10.1007/BF00328141

Uncontrolled Keywords:

Protoplast fusion; Nitrate reductase deficiency; Kanamycin; Nicotiana tabacum; Nicotiana sylvestris

BORIS DOI:

10.7892/boris.101267

URI:

https://boris.unibe.ch/id/eprint/101267

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