Summary
We have investigated the fate of the mitochondrial genomes of cybrids derived from “donor-recipient” protoplast fusion between X-irradiated Raphanus sativus (cms line) and iodoacetamide-treated Brassica napus cv. Westar. Two out of ten fusion products were male-sterile with the diploid chromosome number of B. napus. The mitochondrial (mt) genomes of the cybrids and their progeny were further analyzed by DNA-DNA hybridizaion using the pea mitochondrial ATPase subunit gene (atpA) as a probe. One cybrid, 18-3, had a 3.0 kb fragment characteristic of B. napus and a 2.0 kb non-parental fragment when the BamHI-digested DNA was hybridized with the probe. In the first-backcrossed progeny of this cybrid, the hybridization pattern was not stably inherited. A 4.0 kb radish fragment, not detectable in the cybrid, appeared in one of the BC1 generation siblings, and the 2.0 kb non-parental fragment was lost in another. The hybridization patterns in BC1 progeny siblings of cybrid 12-9 were also varied. The alteration of mtDNA in the cybrid progeny continued to the BC2 generation. There was no clear evidence of a heteroplasmic state or of sub-stoichiometric molecules in the mt genome of cybrid 18-3. A possible cause of the observed alteration in the mt genome is discussed.
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References
Akagi H, Sakamoto M, Negishi T, Fujimura T (1989) Construction of rice cybrid plant. Mol Gen Genet 215:501–506
Aviv D, Galun E (1987) Chondriome analysis in sexual progenies of Nicotiana cybrids. Theor Appl Genet 73:821–826
Barsby TL, Choung PV, Yarrow SA, Wu SC, Coumans M, Kemble RJ, Powell KP (1987) The combination of Polima cms and cytoplasmic triazine resistance in Brassica napus. Theor Appl Genet 73:809–814
Boeshore ML, Lifshitz I, Hanson MR, Izhar S (1983) Novel composition of mitochondrial genomes in Petunia hybrids derived from cytoplasmic male-sterile and fertile plants. Mol Gen Genet 190:459–467
Bonhomme S, Budar F, Férault M, Pelletier G (1991) A 2.5 kb NcoI fragment of Ogura radish mitochondrial DNA is correlated with cytoplasmic male-sterility in Brassica cybrids. Curr Genet 19:121–127
Escote-Carlson LJ, Gabay-Laughnan S, Laughnan JR (1990) Nuclear genotype affects mitochondrial genome organization of cms-S maize. Mol Gen Genet 223:457–464
Fedoroff N, Wellauer PK, Wall R (1977) Intermolecular duplexes in heterogeneous nuclear RNA from Hela cells. Cell 10:567–610
Ichikawa H, Tanno-Suenaga L, Imamura J (1988) Transfer of mitochondria through protoplast fusion. In: Bajaj Y. P. S. (ed) Biotechnology in Agriculture and Forestry, vol 9. Springer-Verlag, Berlin Heidelberg, pp 360–375
Izhar S, Schlicter M, Swartzberg D (1983) Sorting out of cytolasmic elements in somatic hybrids of Petunia and the prevalence of the heteroplasmon through several meiotic cycles. Mol Gen Genet 190:468–474
Jarl CI, Van Grinsven MQJM, Van den Mark F (1989) Correction of chlorophyll-defective male-sterile winter oilseed rape (Brassica napus) through organellar exchange: molecular analysis of the cytoplasm of parental lines and corrected progeny. Theor Appl Genet 77:135–141
Kao HM, Brown GG, Scoles G, Séguin-Swartz G (1991) Ogura cytoplasmic male sterility and triazine tolerant Brassica napus cv. Westar produced by protoplast fusion. Plant Sci 75:63–72
Kumashiro T, Asahi T, Nakaido F (1989) Transfer of cytoplasmic factors by asymmetric fusion from a cross-incompatible species, Nicotiana repanda to N. tabacum, and characterization of cytoplasmic genomes. Plant Sci 61:137–144
Kyozuka J, Kaneda T, Shimamoto K (1989) Production of cytoplasmic male-sterile rice (Oryza sativa L.) by cell fusion. Bio/technology 7:1171–1174
Makaroff CA, Apel IJ, Palmer JD (1990) Characterization of radish mitochondrial atpA: influence of nuclear background on transcription of atpA-associated sequences and relationship with male sterility. Plant Mol Biol 15:735–746
Menczel L, Wolfe K (1984) High frequency of fusion induced in freely suspended protoplast mixtures by polyethylene glycol and dimethylsulfoxide at high pH. Plant Cell Rep 3:196–198
Menczel L, Morgan A, Brown S, Maliga P (1987) Fusion-mediated combination of Ogura-type cytoplasmic male sterility with Brassica napus plastids using X-irradiated cms protoplasts. Plant Cell Rep 6:98–101
Morgan A, Maliga P (1987) Rapid chloroplast segregation and recombination of mitochondrial DNA in Brassica cybrids. Mol Gen Genet 209:240–246
Morikami A, Nakamura K (1987) Structure and expression of pea mitochondrial F1-ATPase a subunit gene and its pseudogene involved in homologous recombination. J Biochem 101:967–976
Namai H, Sarashima M, Hosoda T (1980) Interspecific and intergeneric hybridization breeding in Japan. In: Tunoda S, Hinata K, Gomez-campo C (eds) Brassica crops and wild allies. Japan Scientific Societies Press, Tokyo, pp 191–203
Nishibayashi S, Kaeriyama J (1986) Structural stability of chromosomes in rice (Oryza sativa L.) plants regenerated from somatic tissue culture. Plant Tiss Cult Lett 3:31–34
Pelletier G, Primard C, Vedel F, Chétrit P, Remy R, Rousselle P, Renard M (1983) Intergeneric cytoplasmic hybridization in cruciferae by protoplast fusion. Mol Gen Genet 191:244–250
Rosén B, Halldén C, Heneen WK (1988) Diploid Brassica napus somatic hybrids: characterization of nuclear and organellar DNA. Theor Appl Genet 76:197–203
Sakai T, Imamura J (1990) Intergeneric transfer of cytoplasmic male sterility between Raphanus sativus (cms line) and Brassica napus through cytoplast-protoplast fusion. Theor Appl Genet 80:421–427
Shirzadegan M, Christy M, Earle ED, Palmer JD (1989) Rearrangement, amplification, and assortment of mitochondrial DNA molecules in cultured cells of Brassica campestris. Theor Appl Genet 77:17–25
Shirzadegan M, Palmer JD, Christey M, Earle ED (1991) Patterns of mitochondrial DNA instability in Brassica campestris cultured cells. Plant Mol Biol 16:21–37
Small ID, Issac PG, Leaver CJ (1987) Stoichiometric differences in DNA molecules containing the atpA gene suggest mechanisms for the generation of mitochondrial genome diversity in maize. EMBO J 6(4): 865–869
Small ID, Suffolk R, Leaver CJ (1989) Evolution of plant mitochondrial genomes via sub-stoichiometric intermediates. Cell 58:69–76
Tanno-Suenaga L, Ichikawa H, Imamura J (1988) Transfer of the cms trait in Daucus carota L. by “donor-recipient” protoplast fusion. Theor Appl Genet 76:855–860
Vedel F, Chétrit P, Mathieu C, Pelletier G, Primard C (1986) Several different mitochondrial DNA regions are involved in intergenomic recombination in Brassica napus cybrid plants. Curr Genet 11:17–24
Wang Y, Gengenbach B (1989) Heterogeneity and changes in mitochondrial atpA genes among related cms-S cytoplasmic lines of maize. Maydica 34:217–225
Yang Z-Q, Shikanai K, Mori K, Yamada Y (1989) Plant regeneration from cytoplasmic hybrids of rice (Oryza sativa L.). Theor Appl Genet 77:305–310
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Communicated by Yu. Gleba
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Sakai, T., Imamura, J. Alteration of mitochondrial genomes containing atpA genes in the sexual progeny of cybrids between Raphanus sativus cms line and Brassica napus cv. Westar. Theoret. Appl. Genetics 84, 923–929 (1992). https://doi.org/10.1007/BF00227405
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DOI: https://doi.org/10.1007/BF00227405