Skip to main content
SpringerLink
Log in

Morphological and molecular characterization of somatic hybrid plants between Lycopersicon esculentum and Solanum nigrum

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Mesophyll protoplasts of tomato (Lycopersicon esculentum Mill. var. cerasiforme) and of an atrazine-resistant biotype of black nightshade, (Solanum nigrum L.), were fused by using polyethylene glycol/dimethyl sulfoxide (PEG/DMSO) solution and three somatic hybrid plants, each derived from a separate callus, were recovered. A twostep selection system was used: (1) protoplast culture medium (modified 8E) in which only tomato protoplasts formed calluses; and (2) regeneration medium (MS2Z) on which only S. nigrum calluses produced shoots. These selective steps were augmented by early isozyme analysis of putative hybrid shoots still in vitro. Phosphoglucoisomerase (PGI) and glutamate oxaloacetate transaminase (GOT) mapped to five loci on four chromosomes in tomato confirmed the hybrid nature of the nuclei of regenerated shoots. The somatic hybrid plants had simple leaves, and intermediate flower and bud morphology, but anthesis was reduced to 5% due to premature bud abscission and the pollen grains were non-viable. Southern DNA blot hybridization using a pea 45 S ribosomal RNA gene probe reconfirmed the hybrid nature of the nuclear genome of the three plants. A 32P-labeled probe of Oenothera chloroplast DNA (cpDNA) hybridized to cpDNA restricted with EcoRI or EcoRV indicated the presence of the tomato cpDNA pattern in all three hybrids. Likewise, the plants were all found to be atrazine sensitive. Analysis with two mitochondrial (mt)DNA-specific probes, maize cytochrome oxidase subunit II and PmtSylSa8 from Nicotiana sylvestris, showed that, in addition to typical mitochondrial rearrangements, specific bands of both parents were present or missing in each somatic hybrid plant.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Austin S, Baer MA, Helgeson JP (1985) Transfer of resistance to potato leaf roll virus from Solanum brevidens into Solanum tuberosum by somatic fusion. Plant Sci 39:75–82

    Google Scholar 

  • Aviv D, Arzee-Gonen P, Bleichman S, Galun E (1984) Novel alloplasmic Nicotiana plants by ‘donor-recipient’ protoplast fusion: cybrids having N. tabacum or N. sylvestris nuclear genomes and either or both plastomes and condriomes from alien species. Mol Gen Genet 196:244–253

    Google Scholar 

  • Barsby TL, Shepard JF, Kemble RJ, Wong R (1984a) Somatic hybridization in the genus Solanum tuberosum and S. brevidens. Plant Cell Rep 3:165–167

    Google Scholar 

  • Barsby TL, Yarrow SA, Shepard JF (1984b) Heterokaryon identification through simultaneous fluorescence of tetramethylrhodamine isothiocyanate and fluorescein isothiocyanate labelled protoplasts. Stain Technol 59:217–220

    Google Scholar 

  • Belliard G, Vedel F, Pelletier G (1979) Mitochondrial recombination in cytoplasmic hybrids of Nicotiana tabacum by protoplast fusion. Nature 281:401–403

    Google Scholar 

  • Binding H, Jain SM, Finger J, Mordhorst G, Nehls R, Gressel J (1982) Somatic hybridization of triazine resistant biotype of Solanum nigrum with S. tuberosum. 1. Clonal variations in morphology and atrazine sensitivity. Theor Appl Genet 63:273–277

    Google Scholar 

  • Binding H, Cohen N, Gressel J (1984) Somaclonal variation in potato (x) S. nigrum protoplast fusants. Abstract. International Symposium of Genetic Manipulation in Crops, Beijing, China, p. 99

  • 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

    Google Scholar 

  • Clark E, Schnabelrauch L, Hanson MR, Sink KC (1986) Differential fate of plastid and mitochondrial genomes in Petunia somatic hybrids. Theor Appl Genet 72:748–755

    Google Scholar 

  • Davey MR, Kumar A (1983) Higher plant protoplasts — retrospect and prospect. Int Rev Cytol [Suppl] 16:219–263

    Google Scholar 

  • Fluhr R, Aviv D, Edelman M, Galun E (1983) Cybrids containing mixed and sorted out chloroplasts following interspecific somatic fusion in Nicotiana. Theor Appl Genet 65:289–294

    Google Scholar 

  • Fox TD, Leaver CJ (1981) The Zea mays mitochondrial gene coding cytochrome oxidase subunit II has an intervening sequence and does not contain TGA codons. Cell 26:315–323

    Google Scholar 

  • Frearson EM, Power JB, Cocking EC (1973) The isolation, culture and regeneration of petunia leaf protoplasts. Dev Biol 33:130–137

    Google Scholar 

  • Galun E, Arzee-Gonen P, Fluhr R, Edelman M, Aviv D (1982) Cytoplasmic hybridization in Nicotiana: Mitochondria DNA analysis in prognies resulting from fusion between protoplasts having different organelle constitution. Mol Gen Genet 186:50–56

    Google Scholar 

  • Gleddie S, Keller WA, Setterfield G (1986) Production and characterization of somatic hybrids between Solanum melongena and S. sisymbriifolium Lam. Theor Appl Genet 71:613–621

    Google Scholar 

  • Gounaris I, Michalowski CB, Bohnert HJ, Price CA (1986) Restriction and gene maps of plastid DNA from Capsicum annuum. Curr Genet 11:7–16

    Google Scholar 

  • Gressel J (1985) The molecular anatomy of resistance to photosystem II herbicides. In: Miffin BJ (ed) Oxford surveys of plant molecular and cell biology, vol. 2. Oxford University Press, Oxford, pp 319–328

    Google Scholar 

  • Guri A, Izhar S (1984) Improved efficiency of plant regeneration from protoplasts of eggplant (Solanum melongena L.). Plant Cell Rep 3:247–249

    Google Scholar 

  • Handley LW, Nickels RL, Cameron MW, Moore PP, Sink KC (1986) Somatic hybrid plants between Lycopersicon esculentum and Solanum lycopersicoides. Theor Appl Genet 71:691–697

    Google Scholar 

  • Hanson MR, Boeshore ML, McClean PE, O'Connell MA, Nivison HT (1986) The isolation of mitochondria and mitochondrial DNA. Methods Enzymol 118:437–453

    Google Scholar 

  • Hosticka LP, Hanson MR (1984) Induction of plastid mutation in tomato by nitrosomethyl urea. J Hered 75:242–246

    Google Scholar 

  • Huang A, Liu I, Youle R (1976) Organelle specific isozyme of aspartate α-ketoglutarate transaminase in spinach leaves. Plant Physiol 58:110–113

    Google Scholar 

  • Izhar S, Schlichter M, Swartzberg D (1983) Sorting out in somatic hybrids of Petunia and the prevalence of the heteroplasmon through several meiotic cycles. Mol Gen Genet 190:468–474

    Google Scholar 

  • Kao KN, Michayluk MR (1981) Embryoid formation in alfalfa cell suspension cultures from different plants. In Vitro 17:645–648

    Google Scholar 

  • Kemble RJ, Shepard JF (1984) Cytoplasmic DNA variation in a potato protoclonal population. Theor Appl Genet 69:211–216

    Google Scholar 

  • Kemble RJ, Flavell RB, Brettell RIS (1982) Mitochondrial DNA analysis of fertile and sterile maize plants derived from tissue culture with the Texas male sterile cytoplasm. Theor Appl Genet 62:213–217

    Google Scholar 

  • Kinsara A, Patnaik SN, Cocking EC, Power JB (1986) Somatic hybrid plants of Lycopersicon esculentum Mill. and Lycopersicon peruvianum Mill. J Plant Physiol 125:225–234

    Google Scholar 

  • Lonsdale DN, Hodge TP, Stoehr PH (1986) Analysis of the genome structure of plant mitochondria. Methods Enzymol 118:453–470

    Google Scholar 

  • Melchers G (1982) The first decennium of somatic hybridization. Proceedings 5th Intern Congress Plant Tissue and Cell Culture. Tokyo, Japan, pp 13–18

    Google Scholar 

  • Melchers G, Sacristán MD, Holder AA (1978) Somatic hybrid plants of potato and tomato regenerated from fused protoplasts. Carlsberg Res Commun 43:203–218

    Google Scholar 

  • Menczel L, Wolfe K (1984) High frequency of fusion induced in freely suspended protoplast mixture of polyethylene glycol and dimethylsulfoxide at high pH. Plant Cell Rep 3:196–198

    Google Scholar 

  • Murashige T, Skoog F (1962) Revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Google Scholar 

  • Nagy F, Lazar G, Menczel L, Maliga P (1983) A heteroplasmic state induced by protoplast fusion is a necessary condition for detecting rearrangement in Nicotiana mitochondrial DNA. Theor Appl Genet 66:203–207

    Google Scholar 

  • Negrutiu I, Jacob M, Caboche M (1984) Advances in somatic cell genetics of higher plants — the protoplast approach in basic studies on mutagenesis and isolation of biochemical mutants. Theor Appl Genet 67:289–304

    Google Scholar 

  • Niedz RP, Rutter SM, Handley LW, Sink KC (1985) Plant regeneration from leaf protoplasts of six tomato cultivars. Plant Sci 39:199–204

    Google Scholar 

  • O'Connell MA, Hanson MR (1986) Regeneration of somatic hybrid plants formed between Lycopersicon esculentum and Solanum rickii. Theor Appl Genet 72:59–65

    Google Scholar 

  • Palmer JD, Shields CR (1984) Tripartite structure of the Brassica campestris mitochondrial genome. Nature 307:437–440

    Google Scholar 

  • Rick CM (1983) Tomato (Lycopersicon): In: Tanksley SD, Orton TJ (eds) Isozymes in plant genetic and breeding, part B. Elsevier, Amsterdam, pp 147–165

    Google Scholar 

  • Rothenberg M, Boeshore ML, Hanson MR, Izhar S (1985) Intergenomic recombination of mitochondrial genomes in a somatic hybrid plant. Curr Genet 9:615–618

    Google Scholar 

  • Scandalois JG (1969) Genetic control of multiple molecular forms of enzymes in plants: A review. Biochem Genet 3:37–39

    Google Scholar 

  • Scandalios JG, Sorenson JC, Ott LA (1975) Genetic control and intracellular localization of glutamate oxaloacetic transaminase in maize. Biochem Genet 13:759–769

    Google Scholar 

  • Shepard JF (1980) Abscisic acid-enriched shoot initiation in protoplast-derived calli of potato. Plant Sci Lett 10:335–340

    Google Scholar 

  • Shepard JF, Bidney D, Barsby TL, Kemble RJ (1983) Genetic transfer in plants through interspecific protoplast fusion. Science 219:683–688

    Google Scholar 

  • Tabaeizadeh Z, Pring DR, Vasil IK (1987) Analysis of mitochondrial DNA from somatic hybrid cell lines of Saccharum officinarum (Sugarcane) and Pennisetum americanum (Pearl millet). Plant Mol Biol 8:509–513

    Google Scholar 

  • Tanksley SD, Rick CM (1980) Isozymic gene linkage map of the tomato: applications in genetics and breeding. Theor Appl Genet 57:161–170

    Google Scholar 

  • Uchimiya H, Ohgawara T, Kato H, Akiyama T, Harada H (1983) Detection of two different nuclear genomes in parasexual hybrids by ribosomal RNA gene analysis. Theor Appl Genet 64:117–188

    Google Scholar 

  • Vallejos CE (1983) Enzyme activity staining. In: Tanksley SD, Orton TJ (eds) Isozymes in plant genetics and breeding, part A. Elsevier Amsterdam, pp 496–516

    Google Scholar 

  • Vallejos CE, Tanksley SD (1983) Segregation of isozyme markers and cold tolerance in an interspecific backross of tomato. Theor Appl Genet 66:241–247

    Google Scholar 

  • Vedel F, Chetrit P, Mathieu C, Pelletier G, Primard C (1986) Several different mitochondrial DNA regions are involved in intergenomic recombinations in Brassica napus cybrid plants. Curr Genet 11:17–24

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Horticulture, Michigan State University, 48824, East Lansing, MI, USA

    A. Guri, A. Levi & K. C. Sink

Authors
  1. A. Guri
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Levi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. C. Sink
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by G. Melchers

Michigan Agricultural Experiment Station Journal Article No. 12433

Rights and permissions

Reprints and permissions

About this article

Cite this article

Guri, A., Levi, A. & Sink, K.C. Morphological and molecular characterization of somatic hybrid plants between Lycopersicon esculentum and Solanum nigrum . Mol Gen Genet 212, 191–198 (1988). https://doi.org/10.1007/BF00334683

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00334683

Key words

Search

Navigation