Restriction fragment length polymorphism (RFLP) analyses of plants produced by in vitro anther culture of Solanum chacoense Bitt |
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Authors: | S. R. Rivard M. Cappadocia G. Vincent N. Brisson B. S. Landry |
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Affiliation: | (1) Department of Plant Physiology, Uppsala University, Box 540, S-751 21 Uppsala, Sweden;(2) Department of Genetics, Uppsala University, Box 7003, S-750 07 Uppsala, Sweden |
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Abstract: | Summary Green mesophyll protoplasts of the dihaploid potato line 1982 (Solanum tuberosum L.) were fused with herbicide-bleached mesophyll protoplasts of the dihaploid potato line 679 using a polyethylene glycol protocol. Heterokaryons were identified under a fluorescence microscope using the dual fluorescence of carboxyfluorescein-stained, herbicide-bleached protoplasts and the autofluorescence of green mesophyll protoplasts. About 20% of the protoplasts survived the fusion treatment, and the fusion frequency was 3%–4%. Unfused and fused protoplasts were mass cultured for 6 weeks after which vigorously growing calli were selected and transferred to shoot regeneration medium. Somatic hybrids were identified by a combination of five isozyme markers, and the ploidy level was determined by flow cytometry. Out of 15 calli that regenerated shoots, 6 plants derived from 2 different calli were identified as hexaploid somatic hybrids, while one morphologically deviant plant from a third callus was identified as a mixoploid that had lost some enzyme markers after 4 months of culturing. |
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Keywords: | Solanum tuberosum Protoplast fusion Somatic hybrids Isozymes Ploidy level |
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