Summary
Using the ‘donor-recipient’ protoplast-fusion technique, we have recently constructed several alloplasmic-like lines ofNicotiana in which the original cytoplasms (or part of them) of eitherN. tabacum orN. sylvestris were replaced respectively, either byN. undulata or byN. bigelovii cytoplasms. These cybridizations resulted in two kinds of cytoplasmic male-sterile (CMS) cybrid plants:N. tabacum withN. undulata-like cytoplasm andN. sylvestris withN. bigelovii-like cytoplasm. Fusion of protoplasts, derived from the above two CMS types, by the ‘donor-recipient’ technique, lead to the recovery of 21 cybrid calli. One of these regenerated a cybrid with fertile pollen but having shortened filaments and slighly tappered anthers. Self pollination of the latter cybrid resulted in a second generation progeny having almost normal filaments and anthers. Further selfings produced a third generation in which numerous plants had normal stamens and fertile pollen. Mitochondrial DNA (mtDNA) analysis of second and third generation progenies revealed a novel pattern which differed from each of the parental CMS cybrids and also from the mtDNA of normal, male-fertileNicotiana species. The results suggest that mtDNA recombination between different types of CMS can lead to restoration of male-fertility.
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Aviv D, Arzee-Gonen P, Bleichman S, Galun E: Novel alloplasmicNicotiana plants by ‘Donor-Recipient’ protoplast fusion: cybrids havingN. tabacum orN. sylvestris nuclear genomes and either or both plastomes and chondriomes from alien species. Mol Gen Genet 196: 244–253, 1984.
Aviv D, Fluhr R, Edelman M, Galun E: Progeny analysis of the interspecific somatic hydrids:Nicotiana tabacum (CMS) +Nicotiana sylvestris with respect to nuclear and chloroplast markers. Theor Appl Genet 56: 145–150, 1980.
Aviv D, Galun E: Restoration of fertility in cytoplasmic male-sterile (CMS)Nicotiana sylvestris by fusion with XirradiatedN. tabacum protoplasts. Theor Appl Genet 58: 121–127, 1980.
Aviv D, Galun E: Anin vitro procedure to assign pigment mutations inNicotiana to either the chloroplast or the nucleus. J Hered 76: 135–136, 1985.
Belliard C, Pelletier G, Vedel F, Quetier F: Morphological characteristics and chloroplast DNA distribution in different cytoplasmic parasexual hybrids ofNicotiana tabacum. Mol Gen Genet 165: 231–237, 1978.
Belliard G, Vedel F, Pelletier G: Mitochondrial recombination in cytoplasmic hybrids ofNicotiana tabacum by protoplast fusion. Nature 281: 401–403, 1979.
Boeshore ML, Lifshitz I, Hanson MR, Izhar S: Novel composition of mitochondrial genomes inPetunia somatic hybrids derived from cytoplasmic male sterile and fertile plants. Mol Gen Genet 190: 459–467, 1983.
Chetrit P, Mathieu C, Vedel F, Pelletier G, Primard C: Mitochondrial DNA polymorphism induced by protoplast fusion inCruciferae. Theor Appl Genet 69: 361–366, 1985.
Clark EM, Izhar S, Hanson MR: Independent segregation of the plastid genome and cytoplasmic male sterility inPetunia somatic hybrids. Mol Gen Genet 199: 440–445, 1985.
Clayton DA, Teplitz RL, Nabholz M, Dovey H, Bodmer W: Mitochondrial DNA of human-mouse cell hybrids. Nature 234: 560–562, 1971.
Galun E, Arzee-Gonen P, Fluhr R, Edelman M, Aviv D: Cytoplasmic hybridization inNicotiana: Mitochondrial DNA analysis in progenies resulting from fusion between protoplasts having different organelle constitutions. Mol Gen Genet 186: 50–56, 1982.
Galun E, Aviv D: Cytoplasmic hybridization: genetic and breeding applications. In: Evans DA, Sharp WR, Ammirato PV, Yamada Y (eds) Handbook of Plant Cell Culture. Macmillan Publishing Corp., New York, 1983, pp 358–392.
Galun E, Aviv D: Organelle transfer. Methods in Enzymology 118: 595–611, 1986.
Gerstel DU: Cytoplasmic male sterility inNicotiana (A review). Tech Bull No. 263, 31 pp, North Carolina Agr. Res. Service, 1980.
Glimelius K, Chen K, Bonnett HT: Somatic hybridization inNicotiana: segregation of organellar traits among hybrid and cybrid plants. Planta 153: 504–510, 1981.
Horak I, Coon HG, Dawid IB: Interspecific recombination of mitochondrial DNA molecules in hybrid somatic cells. Proc Natl Acad Sci USA 71: 1828–1832, 1974.
Matthews BF, Widholm JM: Organelle DNA compositions and isoenzyme expression in an interspecific somatic hybrid ofDaucus. Mol Gen Genet 198: 371–376, 1985.
Medgyesy P, Fejes E, Maliga P: Interspecific chloroplast recombination in aNicotiana somatic hybrid. Proc Natl Acad Sci USA 82: 6960–6964, 1985.
Menczel L, Nagy F, Lazar G, Maliga P: Transfer of cytoplasmic male sterility by selection for streptomycin resistance after protoplast fusion inNicotiana. Mol Gen Genet 189: 365–369, 1983.
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497, 1962.
Nagy F, Lazar G, Menczel L, Maliga P: A heteroplasmic state induced by protoplast fusion is a necessary condition for detecting rearragements inNicotiana mitochondrial DNA. Theor Appl Genet 66: 203–207, 1983.
Nagy F, Torok I, Maliga P: Extensive rearrangements in the mitochondrial DNA in somatic hybrids ofNicotiana tabacum andNicotiana knightiana. Mol Gen Genet 183: 437–439, 1981.
Pelletier G, Primand C, Vedel F, Chetrit P, Remy R, Rouselle P, Renard M: Intergeneric cytoplasmic hybridization inCruciferae by protoplast fusion. Mol Gen Genet 191: 244–250, 1983.
Pring DR, Lonsdale DM: Molecular biology of higher plant mitochondrial DNA. International Rev Cytol 97: 1–46, 1985.
Zelcer A, Aviv D, Galun E: Interspecific transfer of cytoplasmic male sterility by fusion between protoplasts of normalNicotiana sylvestris and X-ray irradiated protoplasts of male-sterileN. tabacum. Z Pflanzenphysiol 90: 397–407, 1978.
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Aviv, D., Galun, E. Restoration of male fertileNicotiana by fusion of protoplasts derived from two different cytoplasmic male-sterile cybrids. Plant Mol Biol 7, 411–417 (1986). https://doi.org/10.1007/BF00020325
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DOI: https://doi.org/10.1007/BF00020325