Summary
The chloroplast (cp) and mitochondrial (mt) DNAs of Petunia somatic hybrid plants, which were derived from the fusion of wild-type P. parodii protoplasts with albino P. inflata protoplasts, were analyzed by endonuclease restriction and Southern blot hybridization. Using 32P-labelled probes that distinguished the two parental cpDNAs at a BamH1 site and at a HpaII site, only the P. parodii chloroplast genome was detected in the 10 somatic hybrid plants analyzed. To examine whether cytoplasmic mixing had resulted in rearrangement of the mitochondrial genome in the somatic hybrids, restriction patterns of purified somatic hybrid and parental mtDNAs were analyzed. Approximately 87% of those restriction fragments which distinguish the two parental genomes are P. inflata-specific. Restriction patterns of the somatic hybrid mtDNAs differ both from the parental patterns and from each other, suggesting that an interaction occurred between the parental mitochondrial genomes in the somatic fusion products which resulted in generation of the novel mtDNA patterns. Southern blot hybridization substantiates this conclusion. In addition, somatic hybrid lines derived from the same fusion product were observed to differ in mtDNA restriction pattern, reflecting a differential sorting-out of mitochondrial genomes at the time the plants were regenerated.
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Belliard G, Vedel F, Pelletier G (1979) Mitochondrial recombination in cytoplasmic hybrids of Nicotiana tabacum by protoplast fusion. Nature 218:401–403
Boeshore ML, Hansol MR, Lifshitz I, Izhar S (1983) Novel composition of mitochondrial genomes in Petunia somatic hybrids derived from cytoplasmic male sterile and fertile plants. Mol Gen Genet 190:459–467
Bonnett HT, Glimelius K (1983) Somatic hybridization in Nicotiana: behavior of organelles after fusion of protoplasts from male-fertile and male-sterile cultivars. Theor Appl Genet 65:213–217
Clark EM, Izhar S, Hanson M (1985) Independent segregation of the plastid genome and cytoplasmic male sterility in Petunia somatic hybrids. Mol Gen Genet 199:440–445
Evans DA, Wetter LR, Gamborg OL (1980) Somatic hybrid plants of Nicotiana glauca and N. tabacum obtained by protoplast fusion. Physiol Plant 48:225–230
Galun E, Arzee-Gonen P, Fluhr R, Edelman M, Aviv D (1982) Cytoplasmic hybridization in Nicotiana: Mitochondrial DNA analysis in progenies resulting from fusion between protoplasts having different organelle DNA constitutions. Mol Gen Genet 186:50–56
Hosticka LP, Hanson MR (1984) Induction of plastid mutations in tomatoes by nitrosomethylurea. J Hered 75:242–246
Kirk JTO, Tilney-Bassett RAE (1978) The plastids. Elsevier/North Holland Biomedical Press, New York, Amsterdam
Kumar A, Cocking EC, Bovenberg WA, Kool AJ (1982) Restriction endonuclease analysis of chloroplast DNA in interspecies somatic hybrids of Petunia. Theor Appl Genet 62:377–383
Lonsdale DM, Hodge TP, Fauron CM-R (1984) The physical map and organization of the mitochondrial genome from the fertile cytoplasm of maize. Nucleic Acids Res 12:9249–9261
Maliga P, Nagy F, Xuan LT, Kiss ZsR, Menczel L, Lazar G (1980) Protoplast fusion to study cytoplasmic traits in Nicotiana. In: Advances in protoplast research. Academiai Kiado, Budapest (Hungary), pp 341–345
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratories, Cold Spring Harbor NY
Medgyesy P, Fejes E, Maliga P (1985) Interspecific chloroplast recombination in a Nicotiana somatic hybrid. Proc Natl Acad Sci USA 82:6960–6964
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325
Nagy F, Torok I, Maliga P (1981) Extensive rearrangements in the mitochondrial DNA in somatic hybrids of Nicotiana tabacum and Nicotiana knightiana. Mol Gen Genet 183:437–439
Palmer JD, Shields CR (1984) Tripartite structure of the Brassica campestris mitochondrial genome. Nature 307:437–440
Power JB, Berry SF, Chapman JV, Cocking EC (1979) Somatic hybrids between unilateral cross-incompatible Petunia species. Theor Appl Genet 55:97–99
Rothenberg M, Boeshore ML, Hanson MR, Izhar S (1985) Intergenomic recombination of mitochondrial genomes in a somatic hybrid plant. Curr Genet 9:615–618
Salts Y, Beckman J (1981) Chloroplast DNA preparation from Petunia and Nicotiana. Plant Mol Biol Newslett 2:73–74
Schnabelrauch LS, Kloc-Bauchan F, Sink KC (1985) Expression of nuclear-cytoplasmic genomic incompatibility in interspecific Petunia somatic hybrid plants. Theor Appl Genet 70:57–65
Scowcroft WR, Larkin PJ (1981) Chloroplast DNA assorts randomly in intraspecific somatic hybrids of Nicotiana debneyi. Theor Appl Genet 60:179–184
Shepard JF, Bidney D, Barsby T, Kemble R (1983) Genetic transfer in plants through interspecific protoplast fusion. Science 219:683–688
Sink KC, Power JB, Natarella NJ (1978) The interspecific hybrid Petunia parodii X P. inflata and its relevance to somatic hybridization in the genus Petunia. Theor Appl Genet 53:205–208
Uchimiya H, Murashige T (1974) Evolution of parameters in the isolation of viable protoplasts from cultured tobacco cells. Plant Physiol 54:936–944
Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
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Clark, E., Schnabelrauch, L., Hanson, M.R. et al. Differential fate of plastid and mitochondrial genomes in Petunia somatic hybrids. Theoret. Appl. Genetics 72, 748–755 (1986). https://doi.org/10.1007/BF00266540
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DOI: https://doi.org/10.1007/BF00266540