Abstract
Mesophyll protoplasts of Lycopersicon esculentum Mill. var. cerasiforme (Dunal) Alef, mutant yellow green 6, Rick and protoplasts of a liquid callus culture of the dihaploid strain HH258 of Solanum tuberosum L. were prepared and many fusion products were visible after the protoplasts were incubated together first in the presence of polyethylene glycol and then with a high Ca2+ ion concentration. The protoplasts were transferred to a rich medium and the resultant calli were cultured. Some calli regenerated normal green shoots which were transferred to soil or grafted onto a tomato stock. The subunit polypeptide pattern of ribulose 1,5-bisphosphate carboxylase prepared from leaf material of four regenerated plants was analyzed by isoelectric focusing. The ribulose bisphosphate carboxylase enzyme oligomer in the four plants contained the small subunit products resulting from the expression of both tomato and potato nuclear genes proving these plants to be somatic hybrids between tomato and potato. In three of the four plants the large subunit polypeptides and hence the functional chloroplast DNA were from tomato whereas in the fourth the large subunit and therefore the chloroplast DNA was derived from potato. The plant material was insufficient to establish the chromosome numbers precisely, however counts close to 50 which is near to the expected 48 were obtained for three of the hybrids whereas in the fourth a number close to 72 was observed. In the absence of a selection system against the potato parent, the analysis of ribulose bisphosphate carboxylase provides a convenient marker to demonstrate the hybrid nature of the plants.
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Behnke, M.: Regeneration in Gewebekulturen einiger dihaploiderSolanum tuberosum-Klone. Z. f. Pflanzenzüchtg. 75, 262–265 (1975)
Chan, P.-H. &S. G. Wildman: Chloroplast DNA codes for the primary structure of the large subunit of Fraction 1 protein. Biochim. Biophys. Acta 277, 677–680 (1972)
Chen, K., J. C. Gray &S. G. Wildman: Fraction 1 protein and the origin of polyploid wheats. Science 190, 1304–1306 (1975)
Coen, D. M., J. R. Bedbrook, L. Bogorad &A. Rich: Maize chloroplast DNA fragment encoding the large subunit of ribulose bisphosphate carboxylase. Proc. Nat. Acad. Sci. (USA) 74, 5487–5491 (1977)
Dobberstein, B., G. Blobel &N.-H. Chua: In vitro synthesis and processing of a putative precursor for the small subunit of ribulose-1,5-bisphosphate carboxylase ofChlamydomonas reinhardtii. Proc. Nat. Acad. Sci. (USA) 74, 1082–1085 (1977)
Dudits, D., I. Rasko, G. Hadlaczky &A. Lima-De-Faria: Fusion of human cells with carrot protoplasts induced by polyethylene glycol. Heriditas 82, 121–124 (1976)
Dudits, D., G. Hadlaczky, E. Levi, O. Fejér &G. Lazar: Somatic hybridization ofDaucus carota andD. cepillifolius by protoplast fusion. Theor. Appl. Genet. 51, 127–132 (1977)
Gatenby, A. A. &E. C. Cocking: Polypeptide composition of Fraction 1 protein subunits in the genusPetunia. Plant Sci. Letts. 10, 97–101 (1977)
Gatenby, A. A. &E. C. Cocking: Fraction 1 protein and the origin of the European potato. Plant Sci. Letts. 12, 177–181 (1978)
Gelvin, S., P. Heizmann &S. H. Howell: Identification and cloning of the chloroplast gene coding for the large subunit of ribulose 1,5-bisphosphate carboxylase fromChlamydomonas reinhardtii. Proc. Nat. Acad. Sci. (USA) 74, 3193–3197 (1977)
Gray, J. C., S. D. Kung, S. G. Wildman &S. J. Sheen: Origin ofNicotiana tabacum L. detected by polypeptide composition of Fraction 1 protein. Nature 252, 226–227 (1974)
Gray, J. C., S. D. Kung &S. G. Wildman: Polypeptide chains of the large and small subunits of Fraction 1 protein from tobacco. Archiv. Biochem. Biophys. 185, 272–281 (1978)
Highfield, P. E. &R. J. Ellis: Synthesis and transport of the small subunit of chloroplast ribulose bisphosphate carboxylase. Nature 271, 420–424 (1978)
Holder, A. A.: Ribulose 1,5-diphosphate carboxylase fromOenothera: Purification and a peptide mapping procedure for the subunits. Carlsberg Res. Commun. 41, 321–334 (1976)
Hollingshead, L.: A lethal factor inCrepis effective only in an interspecific hybrid. Genetics 15, 114–140 (1930)
Hollingshead, L.: Cytological investigations of hybrids and hybrid derivatives ofCrepis capillaris andCrepis tectorum. Univ. Calif. Publ. Agri. Sci. 6, 55–94 (1930)
Kahl, G. &M. Müller: Phenol induced inactivation of phosphoglucomutase in extracts from different organs of the potato plant. Biochem. Physiol. Pflanzen 169, 281–288 (1976)
Kao, K. N.: Chromosomal behavior in somatic hybrids of soybean—Nicotiana glauca. Molec. gen. Genet. 150, 225–230 (1977)
Kao, K. N. &M. R. Michayluk: A method for high frequency intergeneric fusion of plant protoplasts. Planta (Berlin) 115, 355–367 (1974)
Kao, K. N. &M. R. Michayluk: nutritional requirements for growth ofVicia hajastana cells and protoplasts at very low density in liquid media. Planta (Berlin) 126, 105–110 (1975)
Kawashima, N. &S. G. Wildman: Studies on Fraction 1 protein. IV. Mode of inheritance of the primary structure in relation to whether chloroplast or nuclear DNA contains the code for a chloroplast protein. Biochim. Biophys. Acta 262, 42–49 (1972)
Keller, W. A. &G. Melchers: The effect of high pH and calcium on tobacco leaf protoplast fusion. Z. Naturforsch. 28c, 737–741 (1973)
Kung, S. D.: Tobacco Fraction 1 protein: a unique genetic marker. Science 191, 429–434 (1976)
Kung, S. D., J. C. Gray &S. G. Wildman: Polypeptide composition of Fraction 1 protein from parasexual hybrid plants in the genusNicotiana. Science 187, 353–355 (1975)
Maliga, P., Z. R. Kiss, A. H. Nagy &G. Lazar: Genetic instability in somatic hybrids ofNicotiana tabacum andNicotiana knightiana. Molec. gen. Genet. 163, 145–152 (1978)
Melchers, G.: Genetik und Evolution. Bericht eines Botanikers. Z. Vererbungslehre 76, 229–259 (1939)
Melchers, G.: Microbial techniques in somatic hybridisation by fusion of protoplasts. In: Internat. Cell Biol. 1976–77, B. R. Brinkley and K. R. Porter, eds. Papers presented at the I. Internat. Congress on Cell Biology, Boston Mass. 1976. The Rockefeller University Press 1977, pp. 207–215.
Melchers, G.: Potatoes for combined somatic and sexual breeding methods; plants from protoplasts and fusion of protoplasts of potato and tomato. In: Production of natural compounds by cell culture methods. A. W. Alfermann and E. Reinhard, eds. Proc. of an Internat. Symp. on plant cell culture. Ges. f. Strahlen- und Umweltforschung, München, pp. 306–311 (1978)
Melchers, G. &G. Labib: Somatic hybridisation of plants by fusion of protoplasts. I. Selection of light resistant hybrids of “haploid” light sensitive varieties of tobacco. Molec. gen. Genet. 135, 277–294 (1974)
Melchers, G. &M. D. Sacristán: Somatic hybridisation of plants by fusion of protoplasts. II. The chromosome numbers of somatic hybrid plants of 4 different fusion experiments. La Culture des Tissues et de Cellules des Végétaux, Travaux dédiés à la mémoire de Georges Morel, R. Gautheret, ed., Masson, Paris, pp. 169–177 (1977)
Pogliaga, H. H.: Hibrido intergenerico “Nicotiana × Petunia”. Rev. Argent. Agron. 19, 171–178 (1952)
Power, J. B., E. M. Frearson, C. Hayward, D. George, P. K. Evans, S. F. Berry &E. C. Cocking: Somatic hybridisation ofPetunia hybrida andP. parodii Nature 263, 500–502 (1976)
Roy, H., R. Patterson &A. T. Jagendorf: Identification of the small subunit of ribulose bisphosphate carboxylase as a product of wheat leaf cytoplasmic ribosomes. Archiv. Biochem. Biophys. 172, 64–73 (1976)
Sacristán, M. D. &G. Melchers: The caryological analysis of plants regenerated from tumorous and other callus cultures of tobacco. Molec. gen. Genet. 105, 317–333 (1969)
Sakano, K., S. D. Kung &S. G. Wildman: Identification of several chloroplast DNA genes which code for the large subunit of Nicotiana Fraction 1 proteins. Molec. gen. Genet. 130, 91–97 (1974)
Schieder, O.: Fusionen zwischen Protoplasten vonSpaerocarpos donnellii Aust.-Mutanten. Biochem. Physiol. Pflanzen 165, 433–435 (1974)
Schieder, O.: Somatic hybrids ofDatura innoxia Mill.+Datura discolor Bernh. and ofDatura innoxia Mill.+Datura stramonium L. var. tatula L. I. Selection and Characterisation. Molec. gen. Genet. 162, 113–120 (1978)
Smith, H. H., K. N. Kao &N. C. Combatti: Interspecific hybridization by protoplast fusion inNicotiana. J. Hered. 67, 123–128 (1976)
Steer, M. W. &D. Kernoghan: Nuclear and cytoplasmic genome relationships in the genusAvena: Analysis by isoelectric focusing of ribulose bisphosphate carboxylase subunits. Biochem. Genet. 15, 273–286 (1977)
Takebe, I., G. Labib &G. Melchers: Regeneration of whole plants from isolated mesophyll protoplasts of tobacco. Naturwissenschaften 58, 318–320 (1971)
Vesterberg, O.: Isoelectric focusing of proteins in thin layers of polyacrylamide gel. Sci. Tools 20, 22–29 (1973)
Vesterberg, O., L. Hansen &A. Siösten: Staining of proteins after isoelectric focusing in gels by new procedures. Biochim. Biophys. Acta 491, 160–166 (1977)
Wallin, A., K. Glimelius & T. Eriksson: Effects of polyethyleneglycol on aggregation and fusion ofDaucus carota protoplasts. In: Third International Congress of Plant Tissue and Cell Culture, Abstr. No. 207 (1974)
Wetter, L. R.: Isoenzyme patterns in soybean-Nicotiana somatic hybrid cell lines. Molec. gen. Genet. 150, 231–236 (1977)
Zenkteler, M. &G. Melchers: In vitro hybridization by sexual methods and by fusion of somatic protoplasts. Theor. Appl. Genet. 52, 81–90 (1978)
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Melchers, G., Sacristán, M.D. & Holder, A.A. Somatic hybrid plants of potato and tomato regenerated from fused protoplasts. Carlsberg Res. Commun. 43, 203–218 (1978). https://doi.org/10.1007/BF02906548
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DOI: https://doi.org/10.1007/BF02906548