Summary
Direct selection of cybrids by simultaneous selection for “donor” chloroplasts and for the “recipient” nuclei is described. Mesophyll protoplasts of two tobacco (Nicotiana tabacum) mutants, SR1 (streptomycin resistant) and Valr-2 (valine resistant), were fused by polyethylene glycol treatment. Streptomycin resistance in the SR1 mutant is a maternally inherited chloroplast trait while valine resistance is a Mendelian (nuclear) digenic recessive character. The fused protoplast population was cultured and colonies were selected for resistance to valine (1 mM) and streptomycin (343 μM). The efficiency of selection has been confirmed in three clones by demonstrating seed transmission of both streptomycin and valine resistances. In one subclone both streptomycin resistant and sensitive plants were obtained indicating that the streptomycin sensitive chloroplasts had not been totally eliminated by growth on the selective medium.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Belliard G, Pelletier G, Vedel F, Quétier F (1978) Morphological characteristics and chloroplast DNA distribution in different cytoplasmic parasexual hybrids of Nicotiana tabacum. Mol Gen Genet 165:231–237
Bourgin JP (1978) Valine-resistant plants from in vitro selected tobacco cells. Mol Gen Genet 161:225–230
Bourgin JP, Chupeau Y, Missonier C (1979) Plant regeneration from mesophyll protoplasts of several Nicotiana species. Physiol Plant 45:288–292
Bourgin JP, Hommel MC, Missonier C (1980) Expression of resistance to valine in protoplast-derived cells of tobacco mutants. In: Sala F, Parisi B, Cella R, Ciferri O (eds) Plant cell cultures: results and perspectives. Elsevier/North Holland Biomedical Press, Amsterdam, pp 167–177
Bourgin JP, Chupeau MC, Missonier C (1982) Amino acid-resistant plants from tobacco cells selected in vitro. In: Earle ED, Demarly Y (eds) Variability in plants regenerated from tissue culture. Praeger, New York, pp 163–174
Bourgin JP, Goujaud J, Missonier C, Pethe C (1985) Valine resistance, a potential marker in plant cell genetics. 1. Distinction of two types of valine-resistant tobacco mutants isolated from protoplast-derived cells. Genetics 109:393–407
Caboche M (1980) Nutritional requirements of protoplast-derived, haploid tobacco cells grown at low cell densities in liquid medium. Planta 149:7–18
Chupeau Y, Missonier C, Hommel MC, Goujaud J (1978) Somatic hybrids of plants by fusion of protoplasts. Observations on the model system “Nicotiana glauca — Nicotiana langsdorffii”. Mol Gen Genet 165:239–245
Cséplö A, Maliga P (1982) Lincomycin resistance, a new type of maternally inherited mutation in Nicotiana plumbaginifolia. Curr Genet 6:105–109
Cséplö A, Nagy F, Maliga P (1984) Interspecific protoplast fusion to rescue a cytoplasmic lincomycin resistance mutation into fertile Nicotiana plumbaginifolia plants. Mol Gen Genet 198:7–11
Fluhr R, Aviv D, Edelman M, Galun E (1983) Cybrids containing mixed and sorted-out chloroplasts following interspecific somatic fusions in Nicotiana. Theor Appl Genet 65:289–294
Glimelius K, Bonnett HT (1981) Somatic hybridization in Nicotiana: restoration of photoautotrophy to an albino mutant with defective plastids. Planta 153:497–503
Grandbastien MA, Bourgin JP, Caboche M (1985) Valine resistance, a potential marker in plant cell genetics. 2. Optimization of UV-mutagenesis and selection of valineresistant colonies derived from tobacco mesophyll protoplasts. Genetics 109:409–425
Maliga P, Sz-Breznovits A, Márton L (1973) Streptomycin-resistant plants from callus culture of haploid tobacco. Nature New Biol 244:29–30
Maliga P, Sz-Breznovits A, Márton L, Joo F (1975) Non-Mendelian streptomycin-resistant tobacco mutant with altered chloroplasts and mitochondria. Nature 255:401–402
Maliga P, Lörz H, Lázár G, Nagy F (1982) Cytoplast-protoplast fusion for interspecific chloroplast transfer in Nicotiana. Mol Gen Genet 185:211–215
Medgyesy P, Menczel L, Maliga P (1980) The use of cytoplasmic streptomycin resistance: chloroplast transfer from Nicotiana tabacum into Nicotiana sylvestris, and isolation of their somatic hybrids. Mol Gen Genet 179:693–698
Menczel L, Galiba 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, Nagy F, Lázár G, Maliga P (1983) Transfer of cytoplasmic male sterility by selection for streptomycin resistance after protoplast fusion in Nicotiana. Mol Gen Genet 189:365–369
Meyer Y, Abel WO (1975) Budding and cleavage division of tobacco mesophyll protoplasts in relation to pseudo-wall and wall formation. Planta 125:1–13
Roth EJ, Lark KG (1984) Isopropyl-N-(3-chlorophenyl) carbamate (CIPC) induced chromosomal loss in soybean: a new tool for plant somatic cell genetics. Theor Appl Genet 68:421–431
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
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
Bourgin, J.P., Missonier, C. & Goujaud, J. Direct selection of cybrids by streptomycin and valine resistance in tobacco. Theoret. Appl. Genetics 72, 11–14 (1986). https://doi.org/10.1007/BF00261447
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00261447