Protoplast Fusion and Generation of Somatic Hybrids
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Protoplast
Somatic fusion
Discoveries in the 1960s and 1970s showed that plant protoplasts could be freed from their walls by digestion with fungal enzymes (), grown in culture, and regenerated back into intact plants (). This work opened the way for protoplast fusion and somatic hybridization in plants (). In the ensuing 20 years, there have been substantial improvements in protoplast fusion and culture, and sophisticated approaches for the selection and characterization of somatic hybrids have been developed. As a result, in the 1990s, protoplast fusion has come of age, and fusion-derived somatic hybrids are being evaluated in practical plant breeding programs (, , .
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To transfer only part of genome, X-ray irradiated suspension cell protoplasts of N. langsdorffii were fused with suspension cell protoplasts of N. glauca by polyethylene glycol. Somatic hybrid calli were selected by the growth in the hormone-free medium. Some of somatic hybrid calli from fusion with irradiated protoplasts indicated the loss of small subunit polypeptide of fraction 1 protein which was coded by N. langsdorffii nuclear DNA. Cytological analysis provided an information on significant decrease of chromosomes in somatic hybrid calli from fusion with irradiated protoplasts, compared with the somatic hybrid calli from fusion with non-irradiated protoplasts. In addition, isozyme analysis revealed that somatic hybrid calli from fusion with irradiated protoplasts lost particular bands of N. langsdorffli. These results demonstrate the tranfer of only part of genome from N, langsdorffii to N, glauca by fusion with X-ray irradiated protoplasts.
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Cell fusion
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Protoplast
Somatic fusion
Heterokaryon
Auxotrophy
Cell fusion
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Introduction Why Fuse Isolated Plant Protoplasts? Procedures for Protoplast Fusion Chemical Fusion of Protoplasts Electrical Fusion of Protoplasts Products of Protoplast Fusion and the Selection of Somatic Hybrid Tissues and Plants Somatic Hybrid Tissues and Plants Homokaryons and Heterokaryons Procedures for Selecting Hybrid Cells and Plants Characterization of Somatic Hybrid Plants Generation of Asymmetrical Hybrids and Cybrids by Protoplast Fusion Gametosomatic Hybridization Transfer of Unique Traits by Protoplast Fusion Concluding Remarks Bibliography
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Somatic fusion
Heterokaryon
Cell fusion
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Somatic fusion
Cell fusion
Plant cell
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Mesophyll protoplasts of Nicouana plwnbaginifoJia (PP) and N. sylvestris (SS) were mixed at equal density, treated with polyethylene glycol (PEG) and then cultured in medium with no selection pressure against parental cells. Cytological, morphological, and biochemical studies showed that about 78%of plants regenerated from protoplast fusion were somatic hybrids. The ploidy levels of somatic hybrids varied, and the frequencies of plants with PPSS, PPPPSS, and PPPPSSSS genome constitutions were 84.8%, 8.7%, and 6.5%, respectively. Aneuploid numbers and chromosome structural changes were common in somatic hybrids. The- successful recovery of somatic hybrids in the absence of artificial selection is attributed to a high viability of the fusion products following PEG treatment and a fast rate of growth and development of hybrid cells during in vitro culture.
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Somatic fusion
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