Abstract
Protoplasts of a light sensitive plastome mutant of Nicotiana tabacum (2 n=48) were irradiated and fused with iodoacetate-treated Nicotiana plumbaginifolia (2 n=20) protoplasts. Treated parental protoplasts were unable to divide. Metabolic complementation, however, helped the recovery of interspecific fusion products which survived and formed calli. Altogether 40 clones were investigated. N. plumbaginifolia plants were obtained in 15 clones (38%), somatic hybrids in 23 clones, and both types of regenerates were found in 2 clones. Irradiation therefore significantly increased the frequency of segregant formation with the non-irradiated N. plumbaginifolia nuclei (the frequency was 1.4% in the absence of irradiation). Regenerated plants in most cases (31 out of 34) contained chloroplasts from the irradiated parent. In 6 clones plants were obtained with both types of chloroplast. Thus, irradiated N. tabacum chloroplasts had an improved chance of dominating the heterokaryonderived cells, many of which contained N. plumbaginifolia nucleus. The system described should be generally applicable for the transfer of chloroplasts without the use of selectable genetic markers.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Atchison, B.A., Whitfeld, P.R., Bottomley, W. (1976) Comparison of chloroplast DNAs by specific fragmentation with EcoRI endonuclease. Mol. Gen. Genet. 148, 263–269
Aviv, D., Galun, E. (1980) Restoration of fertility in cytoplasmic male sterile (CMS) Nicotiana sylvestris by fusion with X-irradiated N. tabacum protoplasts. Theor. Appl. Genet. 58, 121–127
Aviv, D., Fluhr, R., Edelman, M., Galun, E. (1980) Progeny analysis of the interspecific somatic hybrids: Nicotiana tabacum (CMS) +Nicotiana sylvestris with respect to nuclear and chloroplast markers. Theor. Appl. Genet. 56, 145–150
Belliard, G., Pelletier, G., Vedel, F., Quetier, F. (1978) Morphological characteristics and chloroplast DNA distribution in different cytoplasmic parasexual hybrids of Nicotiana tabacum. Mol. Gen. Genet. 165, 231–237
Belliard, G., Vedel, F., Pelletier, G. (1979) Mitochondrial recombination in cytoplasmic hybrids of Nicotiana tabacum by protoplast fusion. Nature (London) 281, 401–403
Brewbaker, J.L., Upadhya, M.D., Makinen, Y., MacDonald, T. (1968) Isoenzyme polymorphism in flowering plants III. Gel electrophoretic methods and applications. Physiol. Plant. 21, 930–940
Chen, K., Wildman, S.G., Smith, H.H. (1977) Chloroplast DNA distribution in parasexual hybrids as shown by polypeptide composition of fraction I protein. Proc. Natl. Acad. Sci. USA 74, 5109–5112
Cocking, E.C. (1980) Protoplasts: past and present. In: Advances in Protoplast Research, pp. 3–15, Ferenczy, L., Farkas, G.L. eds. Akadémiai Kiadó, Budapest
Gerstel, U. (1980) Cytoplasmic male sterility in Nicotiana. North Carolina Tech. Bull. 263, 1–31
Gleba, Y.Y., Butenko, R.H., Sytnik, K.M. (1975) Protoplast fusion and parasexual hybridization in Nicotiana tabacum. Dokl. Acad. Nauk, USSR 221, 1196–1198
Gleba, Y.Y. (1978a) Extranuclear inheritance investigated by somatic hybridization. In: Fronties of plant tissue culture 1978, pp. 95–102, Thorpe, T.A. ed., Univ. of Calgary Press, Calgary, Alberta
Gleba, Y.Y. (1978b) Microdroplet culture: tobacco plants from single mesophyll protoplasts. Naturwissenschaften 65, 158–159
Kao, K.N. (1977) Chromosomal behaviour in somatic hybrids of soybean-Nicotiana glauca. Mol. Gen. Genet. 150, 225–230
Keller, W.A., Melchers, G. (1973) The effect of high pH and calcium on tobacco leaf protoplast fusion. Z. Naturforsch. 28c, 737–741
Linsmaier, E.M., Skoog, F. (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol. Plant. 18, 100–127
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., Nagy, F., Kiss, Zs., Maliga, P. (1981) Streptomycin resistant and sensitive hybrids of Nicotiana tabacum+Nicotiana knightiana: correlation of resistance with N. tabacum plastids. Theor. Appl. Genet. 59, 191–195
Raveh, D., Huberman, E., Galun, E. (1973) In vitro culture of tobacco protoplasts: use of feeder layer techniques to support division of cells plated at low densities. In Vitro 9, 216–222
Schieder, O., Vasil, I.K. (1980) Protoplast fusion and somatic hybridization. Int. Rev. Cytol. Suppl. 11 B, 21–42
Wettstein, D.V., Erikson, G. (1965) The genetics of chloroplasts. In: Genetics today, Proc. 11th Int. Congr. Genet., Vol. 3, pp. 591–610, Geerts, S.J. ed., Pergamon Press, Oxford
Wildman, S.G., Lui-Liao, C., Wong-Staal, F. (1973) Maternal inheritance, cytology and macromolecular composition of defective chloroplasts in variegated mutant of Nicotiana tabacum. Planta 113, 293–312
Wong-Staal, F., Wildman, S.G. (1973) Identification of a mutation in chloroplast DNA correlated with formation of defective chloroplasts in a variegated mutant of Nicotiana tabacum. Planta 113, 313–326
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
Rights and permissions
About this article
Cite this article
Sidorov, V.A., Menczel, L., Nagy, F. et al. Chloroplast transfer in Nicotiana based on metabolic complementation between irradiated and iodoacetate treated protoplasts. Planta 152, 341–345 (1981). https://doi.org/10.1007/BF00388259
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00388259