Abstract
A double-mutant cell line, which was unable to grow in a medium with NO -3 as the sole nitrogen source and was resistant to 5-methyl-tryptophan (5MT), was selected from cell suspensions of Sinapis turgida Del. (Brassicaceae) by culturing the cells in AA medium (Toriyama and Hinata, 1985, Plant Sci. 41, 179–183) supplemented with 50 mM chlorate and 229 μM 5MT. Protoplasts of this cell line were fused with mesophyll protoplasts of Brassica oleracea L. with dextran, and six somatic hybrids were selected initially by culture in the NO -3 medium and then by transfer to the NO -3 medium supplemented with 229 μM 5MT. The somatic hybrids produced embryoids, leaves and plantlets on a regeneration medium. The hybrid characters were confirmed by investigations of acid phosphatase (EC 3.1.3.2) and peroxidase (EC 1.11.1.7) isoenzymes, chromosome number, growth on NO -3 medium, 5MT resistance, and capacity to regenerate plants. Somatic hybrids between S. turgida Del. and B. nigra (L.) Koch were also obtained using this method. These results indicate that the double-mutant cell line established here will be able to serve as a universal hybridizer to select somatic hybrids after protoplast fusion with any other wild-type partner.
Similar content being viewed by others
Abbreviations
- B5:
-
medium of Gamborg et al. (1968)
- MS:
-
medium of Murashage and Skoog (1962)
- 5MT:
-
5-thethyltryptophan
References
Dudits, D., Féjér, O., Hadlaczky, G., Koncz, C., Lázár, G.B., Horváth, G. (1980) Intergeneric gene transfer mediated by plant protoplast fusion. Mol. Gen. Genet. 179, 283–288
Dudits, D., Hadlaczky, G.Y., Bajszár, G.Y., Koncz, C.S., Lázár, G., Horváth, G. (1979) Plant regeneration from intergeneric cell hybrids. Plant Sci. Lett. 15, 101–112
Gamborg, O.L., Miller, R.A., Ojima, K. (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50, 151–158
Gleba, Y.Y., Hoffmann, F. (1978) Hybrid cell lines Arabidopsis thaliana + Brassica campestris: No evidence for specific chromosome elimination. Mol. Gen. Genet. 165, 257–264
Gleba, Y.Y., Hoffmann, F. (1980) “Arabidobrassica”: A novel plant obtained by protoplast fusion. Planta 149, 112–117
Gleba, Y.Y., Momot, V.P., Cherep, N.N., Skarzynskaya, M.V. (1982) Intertribal hybrid cell lines of Atropa belladonna (X) Nicotiana chinensis obtained by cloning individual protoplast fusion products. Theor. Appl. Genet. 62, 75–79
Glimelius, K., Eriksson, T., Grafe, R., Muller, A.J. (1978) Somatic hybridization of nitrate reductase-deficient mutants of Nicotiana tabacum by protoplast fusion. Physiol. Plant. 44, 273–277
Gupta, P.P., Gupta, M., Schieder, O. (1982) Correction of nitrate reductase defect in auxotrophic plant cells through protoplast-mediated intergeneric gene transfers. Mol. Gen. Genet. 188, 378–383
Harberd, D.J. (1976) Cytotaxonomic studies of Brassica and related genera. In: The biology and chemistry of the Cruciferae, pp. 47–68, Vaughan, J.G., Macleod, A.J., Jones, B.M.G., eds. Academic Press, London
Harms, C.T., Potrykus, I., Widholm, J.M. (1981) Complementation and dominant expression of amino acid analogue resistance markers in somatic hybrid clones from Daucus carota after protoplast fusion. Z. Pflanzenphysiol. 101, 377–390
Hoffmann, F., Adachi, T. (1981) “Arabidobrassica”: Chromosomal recombination and morphogenesis in asymmetric intergeneric hybrid cells. Planta 153, 586–593
Horn, M.E., Kameya, T., Brotherton, J.E., Widholm, J.M. (1983) The use of amino acid analog resistance and plant regeneration ability to select somatic hybrids between Nicotiana tabacum and N. glutinosa. Mol. Gen. Genet. 192, 235–240
Kameya, T., Horn, M.E., Widholm, J.M. (1981) Hybrid shoot formation from fused Daucus carota and D. capillifolius protoplasts. Z. Pflanzenphysiol. 104, 459–466
King, J., Khanna, V. (1980) A nitrate reductase-less variant isolated from suspension cultures of Datura innoxia (Mill.). Plant Physiol. 66, 632–636
Krumbiegel, G., Schieder, O. (1979) Selection of somatic hybrids after fusion of protoplasts from Datura innoxia Mill. and Atropa belladonna L. Planta 145, 371–375
Krumbiegel, G., Schieder, O. (1981) Comparison of somatic and sexual incompatibility between Datura innoxia and Atropa belladonna. Planta 153, 466–470
LoSchiavo, F., Giovinazzo, G., Terzi, M. (1983) 8-Azaguanine resistant carrot cell mutants and their use as universal hybridizers. Mol. Gen. Genet. 192, 326–329
Melchers, G., Sacristán, M.D., Holder, A.A. (1978) Somatic hybrid plants of potato and tomato regenerated from fused protoplasts. Carlsberg Res. Commun. 43, 203–218
Müller, A.J., Grafe, R. (1978) Isolation and characterization of cell lines of Nicotiana tabacum lacking nitrate reductase. Mol. Gen. Genet. 161, 67–76
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tabacco tissue cultures. Physiol. Plant. 15, 473–497
Nagao, T. (1982) Somatic hybridization by fusion of protoplasts. III. Somatic hybrids of sexually incompatible combinations Nicotiana tabacum + Nicotiana repanda and Nicotiana tabacum + Salpiglossis sinuata. Jpn. J. Crop Sci. 51, 35–42
Okazaki, K., Hinata, K. (1984) Analysis of S-alleles and S-glycoproteins in F1-hybrid varieties of Japanese radish (Raphanus sativus L.). Jpn. J. Breed. 34, 237–245
Pental, D., Hamill, J.D., Cocking, E.C. (1984) Somatic hybridization using a double mutant of Nicotiana tabacum. Heredity 53, 79–83
Tanabe, T. (1983) Western blotting. Cell Technol. 12, 1061–1068
Toriyama, K., Hinata, K. (1985) Cell suspension and protoplast culture in rice. Plant Sci. 41, 179–183
Tsunoda, S., Hinata, K., Gómez-Campo, C., eds. (1980) Brassica crops and wild allies. Japan Scientific Societies Press, Tokyo
White, D.W.R., Vasil, I.K. (1979) Use of amino acid analogue-resistant cell lines for selection of Nicotiana sylvestris somatic cell hybrids. Theor. Appl. Genet. 55, 107–112
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Toriyama, K., Kameya, T. & Hinata, K. Selection of a universal hybridizer in Sinapis turgida Del. and regeneration of plantlets from somatic hybrids with Brassica species. Planta 170, 308–313 (1987). https://doi.org/10.1007/BF00395021
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00395021