Summary
Protoplasts isolated from a 5-methyltryptophan resistant nonregenerable Nicotiana tabacum (L.) Xanthi suspension culture were fused via the dextran method (Kameya et al. 1981) with protoplasts isolated from N. glutinosa L. leaf mesophyll cells. Prospective somatic hybrids were selected by their ability to produce chlorophyll and to grow on medium containing 5-methyltryptophan and eventually to regenerate into complete plants. Twenty-eight plants regenerated from selected colonies were classified into three groups: (1) Four plants had intermediate morphology, were sterile, had hybrid isozyme patterns for four different enzymes, and contained fraction 1 protein large subunit patterns indicative of a N. glutinosa chloroplast origin. (2) One plant had N. tabacum morphology, was fertile, had two hybrid isozyme patterns, and contained fraction 1 protein large subunits indicative of the N. tabacum chloroplast type. Although this hybrid contained both parental small subunit patterns of fraction 1 protein, two-dimensional gel electrophoresis showed the N. glutinosa-type small subunit to consist of several molecular weights suggestive of errors in the processing steps. Progeny of the Type 2 hybrid (selfed) contained no N. glutinosa characteristics. (3) Twenty-three plants were N. glutinosa escapes. Chromosome numbers ranged from 60 (some chromosome loss) in two hybrids to 34–35 (extensive chromosome loss) in three hybrids.
These results suggest that amino acid analog resistance and regeneration capability can be utilized as complementing markers for selecting and identifying somatic hybrids after protoplast fusion. Mesophyll protoplasts from N. glutinosa can also be regenerated into morphologically normal fertile plants.
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Abbreviations
- 5MT:
-
DL-5-methyltryptophan
- GOT:
-
glutamate oxaloacetate transaminase
- LAP:
-
leucine aminopeptidase
- PER:
-
peroxidase
- RuBPCase:
-
ribulose bisphosphate carboxylase
- PVP:
-
polyvinylpyrolidone
- IEF:
-
isoelectric focusing
- PVPP:
-
polyvinylpolypyrolidone
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Communicated by R.B. Goldberg
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Horn, M.E., Kameya, T., Brotherton, J.E. et al. The use of amino acid analog resistance and plant regeneration ability to select somatic hybrids between Nicotiana tabacum and N. glutinosa . Molec Gen Genet 192, 235–240 (1983). https://doi.org/10.1007/BF00327672
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DOI: https://doi.org/10.1007/BF00327672