Summary
Selection and screening methods were devised which resulted in the identification of a number of somatic hybrid callus clones following fusion of Lycopersicon esculentum protoplasts and L. pennellii suspension culture protoplasts. Visual selection for callus morphology combined with a high fusion frequency and irradiation of one parental protoplast type (137Cs source, 1.5 Krads) resulted in selection of a callus clone population containing a high proportion of somatic hybrids. Analysis of a dimeric isozyme for the presence of a heterodimeric form was found to be satisfactory for distinguishing parental-type calli, somatic hybrid calli, and mixed calli derived from both types of unfused parental cells. No somatic hybrid calli produced shoots, although the sexual hybrid between L. esculentum and L. pennellii regenerated well under the culture conditions employed. This result suggests that the non-regenerable growth habit of the L. pennellii suspension culture was dominant in the somatic hybrid. The culture conditions described here are suitable for obtaining regenerated plants from L. esculentum mesophyll protoplasts. L. esculentum protoplast calli from fusion cultures gave rise to shoots with L. esculentum phenotype at higher frequency than calli from control unfused L. esculentum mesophyll protoplast cultures. The use of probes for species-specific organelle DNA fragments allowed identification of organelle DNA restriction fragments in digests of total DNA from small samples of individual callus clones. The callus clones analyzed either carried predominantly one parental plastid DNA type or mixtures of both types. Use of a mitochondrial DNA (mtDNA) probe which distinguishes two parental mtDNA fragments revealed that the L. pennellii-specific fragment was present in all clones examined, but the L. esculentum fragment was absent or in low proportion.
Similar content being viewed by others
References
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 218: 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
Denhardt D (1976) A membrane filter technique for the detection of complementary DNA. Biochem Biophys Res Commun 23: 641–646
Evans DA, Bravo JE (1983) Protoplast isolation and culture. In: Evans DA, Sharp WR, Ammirato PV, Yamada Y (eds) Handbook of plant cell culture, vol 1. MacMillan, New York, pp 124–176
Galun E, Arzee-Gonen P, Fluhr R, Edelman U, 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
Galun E, Aviv D (1983) Cytoplasmic hybridization: genetics and breeding applications. In: Evans DA, Sharp WR, Ammirato PV, Yamada Y (eds) Handbook of plant cell culture, vol 1. MacMillan, New York, pp 358–392
Gleba Yu Yu, Evans DA (1983) Genetic analysis of somatic hybrid plants. In: Evans DA, Sharp WR, Ammirato PV, Yamada Y (eds) Handbook of plant cell culture, vol 1. MacMillan, New York, pp 322–357
Glimelius K, Bonnett HT (1981) Somatic hybridization in Nicotiana: restoration of photoautotrophy to an albino mutant with defective plastids. Planta 153: 497–503
Gupta PP, Gupta M, Scheider D (1982) Correction of nitrate reductase defect in auxotrophic plant cells through protoplast-mediated intergeneric gene transfers. Mol Gen Genet 188: 378–383
Hanson MR (1982) Cell and tissue culture of Lycopersicon in plant tissue culture. In: Proc 5th Int Cong Plant Tissue Culture. Mt Fuji, Japan, pp 193–194
Hanson MR (1983) Stability variation and recombination in plant mitochondrial genomes via cell culture and somatic hybridization. Oxford Surv Plant Mol Cell Biol 1: 33–52
Harms CT (1983) Somatic hybridization by plant protoplast fusion. Experientia (Suppl) 46: 69–84
Hosticka LP (1983) Master's Thesis, University of Virginia
Koblitz H, Koblitz D (1982) Experiments on tissue culture in the genus Lycopersicon Miller: mesophyll protoplast regeneration to plants in Lycopersicon esculentum cv. ‘Nadja’. Plant Cell Rep 1: 143–146
Maliga P, Lazar G, Joo F, Nagy AH, Menczel L (1977) Restoration of morphogenic potential in Nicotiana by somatic hybridization. Mol Gen Genet 157: 291–296
McClean PE, Hanson MR (submitted) Mitochondrial DNA sequence divergence among Lycopersicon and related Solanum species. Genetics
Medgyesy P, Menczel L, Maliga P (1980) The use of cytoplasmic streptomycin resistance: chloroplast transfer from Nicotiana tabacum into Nicotiana sylvestis, and isolation of their somatic hybrids. Mol Gen Genet 179: 693–698
Melchers G, Sacristan MD, Holder AA (1978) Somatic hybrid plants of potato and tomato regenerated from fused protoplasts. Carlsberg Res Commun 43: 203–218
Menczel L, Nagy F, Kiss Z, Maliga P (1981) Streptomycin resistant and sensitive somatic hybrids of Nicotiana tabacum and Nicotiana knightiana: correlation of resistance to N. tabacum plastids. Theor Appl Genet 59: 191–195
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
Morgan A, Cocking EC (1982) Plant regeneration from protoplasts of Lycopersicon esculentum Mill. Z Pflanzenphysiol 106: 97–104
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucl Acids Res 8: 4321–4325
Nagy F, Török I, Maliga P (1981) Extensive rearrangements in the mitochondrial DNA in somatic hybrids of Nicotiana tabacum and Nicotiana knightiana. Mol Gen Genet 183: 437–439
Nitsch JP (1965) Deux especes photoperiodiques de jours courts: Plumbago indica L. et Plumbago zeylanica. Bull Soc Bot, France 112: 517–522
O'Connell MA, Hanson MR (1984) Somaclonal variation in Lycopersicon regenerates. Plant Physiol 75: 98
Palmer JD, Zamir D (1982) Chloroplast DNA evolution and phylogenetic relationships in Lycopersicon. Proc Natl Acad Sci USA 79: 5006–5010
Rigby P, Dieckmann M, Rhodes C, Berg P (1977) Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113: 237–251
Shepard JF (1981) The isolation culture and regeneration of plant protoplasts. Cold Spring Harbor Course on Plant Molecular Biology
Shepard JF, Bidney D, Barsby T, Kemble R (1983) Genetic transfer in plants through interspecific protoplast fusion. Science 219: 683–688
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517
Tanksley SD (1979) An efficient and economical design for starch gel electrophoresis. Rep Tomato Genet Coop 29: 37–38
Tanksley SD (1980) Pgi-1, a single gene in tomato responsible for a variable number of isozymes. Can J Genet Cytol 22: 271–278
Uchimaya H, Murashige T (1974) Evolution of parameters in the isolation of viable protoplasts from cultured tobacco cells. Plant Physiol 54: 936–944
Author information
Authors and Affiliations
Additional information
Communicated by P. Maliga
Rights and permissions
About this article
Cite this article
O'Connell, M.A., Hanson, M.R. Somatic hybridization between Lycopersicon esculentum and Lycopersicon pennellii . Theoret. Appl. Genetics 70, 1–12 (1985). https://doi.org/10.1007/BF00264476
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00264476