Summary
Protoplasts of Daucus capillifolius isolated from a suspension culture (chromosome number above 60) were X-irradiated over lethal dose (60 krad) just prior to fusion. Protoplasts from D. carota cell line (chromosome number 17) were treated with 15 mM iodoacetamide and fused with the X-irradiated protoplasts. Putative cybrid plants were regenerated on Murashige and Skoog medium (MS) lacking 2,4-D. The regenerated plants possessed chromosome numbers of 17 (2n−1) or 34 (4n−2) and an identical leaf morphology to D. carota. Their mitochondrial DNAs (mtDNAs) were analysed with restriction endonucleases. Novel restriction fragments, not present in mtDNA digests from both parents, were observed in mtDNAs of regenerated plants. These results indicate successful formation of cybrids between D. capillifolius and D. carota by protoplast fusion.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aviv D, Arzee-Gonen P, Bleichman S, Galun E (1984) Novel alloplasmic Nicotiana plants by “donor-recipient” protoplast fusion: cybrids having N. tabacum or N. sylvestris nuclear genomes and either or both plastomes and chondriomes from alien species. Mol Gen Genet 196:244–253
Bayliss MW (1975) The effect of growth in vitro on the chromosome complement of Daucus carota (L.) suspension cultures. Chromosoma 51:401–411
Belliard G, Vedel F, Pelletier G (1979) Mitochondrial re-combination in cytoplasmic hybrids of Nicotiana tabacum by protoplast fusion. Nature 281:401–403
Boeshore ML, Lifshitz I, Hanson MR, 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
Chetrit P, Mathieu C, Vedel F, Pelletier G, Primard C (1985) Mitochondrial DNA polymorphism induced by protoplast fusion in Cruciferae. Theor Appl Genet 69:361–366
Clark E, Schnabelrauch L, Hanson MR, Sink KC (1986) Differential fate of plastid and mitochondrial genomes in Petunia somatic hybrids. Theor Appl Genet 72:748–755
DeBonte LR, Matthews BF (1984) Rapid isolation and purification of plastid and mitochondrial DNA from carrot cell suspensions. Plant Mol Biol Rep 3:32–36
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 constitutions. Mol Gen Genet 186:50–56
Gleba Y, Komarnitsky IK, Kolesnik NN, Meshkene I, Martyn GI (1985) Transmission genetics of the somatic hybridization process in Nicotiana, II. Plastome heterozygotes. Mol Gen Genet 198:476–481
Kao KN, Michayluk MR (1974) A method for high frequency intergeneric fusion of plant protoplasts. Planta 115:355–367
Kothari SL, Monte DC, Widholm JM (1986) Selection of Daucus carota somatic hybrids using drug resistance markers and characterization of their mitochondrial genomes. Theor Appl Genet 72:494–502
Leaver CJ, Gray MW (1982) Mitochondrial genome organization and expression in higher plants. Annu Rev Plant Physiol 33:373–402
Lonsdale DM, Hodge TP, Fauron CM (1984) The physical map and organisation of the mitochondrial genome from the fertile cytoplasm of maize. Nucleic Acids Res 12:9249–9261
Matthews BF, DeBonte LR (1985) Chloroplast and mitochondrial DNAs of the carrot and its wild relatives. Plant Mol Biol Rep 3:12–16
Matthews BF, Widholm JM (1985) Organelle DNA compositions and isoenzyme expression in an interspecific somatic hybrid of Daucus. Mol Gen Genet 198:371–376
Menczel L Galiha G, Nagy F, Maliga P (1982) Effect of radiation dosage on efficiency of chloroplast transfer by protoplast fusion in Nicotiana. Genetics 100:487–495
Menczel L, Polsby L, Steinback KE, Maliga P (1986) Fusion-mediated transfer of triazine-resistant chloroplasts: characterization of Nicotiana tabacum cybrid plants. Mol Gen Genet 205:201–205
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497
Nagy F, Lázár G, Menczel L, Maliga P (1983) A heteroplasmic state induced by protoplast fusion is a necessary condition for detecting rearrangements in Nicotiana mitochondrial DNA. Theor Appl Genet 66:203–207
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
Palmer JD, Shields CR (1984) Tripartite structure of the Brassica campestris mitochondrial genome. Nature 307:437–440
Pelletier G, Primard C, Vedel F, Chetrit P, Remy R, Rousselle P, Renard M (1983) Intergeneric cytoplasmic hybridization in Cruciferae by protoplast fusion. Mol Gen Genet 191:244–250
Sidorov VA, Menczel L, Nagy F, Maliga P (1981) Chloroplast transfer in Nicotiana based on metabolic complementation between irradiated and iodoacetate treated protoplasts. Planta 152:341–345
Steinback KE, McIntosh L, Bogorad L, Arntzen CJ (1981) Identification of the triazine receptor protein as a chloroplast gene product. Proc Natl Acad Sci USA 78:7463–7467
Yarrow SA, Wu SC, Barsby TL, Kemble RJ, Shepard JF (1986) The introduction of CMS mitochondria to triazine tolerant Brassica napus L., var. “Regent”, by micromanipulation of individual heterokaryons. Plant Cell Rep 5:415–418
Zelcer A, Aviv D, Galun E (1978) Interspecific transfer of cytoplasmic male sterility by fusion between protoplasts of normal Nicotiana sylvestris and X-ray irradiated protoplasts of male-sterile N. tabacum. Z Pflanzenphysiol 90:397–407
Author information
Authors and Affiliations
Additional information
Communicated by P. Maliga
Rights and permissions
About this article
Cite this article
Ichikawa, H., Tanno-Suenaga, L. & Imamura, J. Selection of Daucus cybrids based on metabolic complementation between X-irradiated D. capillifolius and iodoacetamide-treated D. carota by somatic cell fusion. Theoret. Appl. Genetics 74, 746–752 (1987). https://doi.org/10.1007/BF00247552
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00247552