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Flow Cytometric Analysis and Sorting of Somatic Hybrid and Transformed Protoplasts

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Book cover Plant Protoplasts and Genetic Engineering II

Part of the book series: Biotechnology in Agriculture and Forestry ((AGRICULTURE,volume 9))

Abstract

Considerable interest has developed in the potential of techniques of flow cytometry and cell sorting for providing insight into the processes of plant growth and development (Brown 1984; Galbraith 1984a, b; Petit et al. 1986; Glimelius 1987). The initial technical advance that culminated in the application of flow cytometry to plant cell systems was the development of procedures for the isolation of protoplasts from a wide variety of plant tissues. This is because a fundamental requirement for the operation of flow cytometers is the availability of suspensions of cells having isometric shapes. As detailed below, flow cytometry involves the analysis of the fluorescence and light-scattering properties of single cells during their passage within a narrow, precisely defined liquid stream through the focus of an intense light source. With the exception of pollen and microspores, protoplasts remain the sole example, within higher plant genera, of single-cell suspensions that are suitable for flow cytometric analysis.

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References

  • Afonso CL, Harkins KR, Thomas-Compton M, Krejci AE, Galbraith DW (1985) Selection of somatic hybrid plants in Nicotiana through fluorescence-activated sorting of protoplasts. Biotechnology 3: 811–816

    Article  Google Scholar 

  • Alexander RG, Cocking EC, Jackson PJ, Jett JH (1985) The characterization and isolation of plant heterokaryons by flow cytometry. Protoplasma 128: 52–58

    Article  Google Scholar 

  • Ayres NM (1987) Inheritance of organelles in somatic cell hybrids and cybrids of Nicotiana. PhD Thesis, Univ Nebraska, Lincoln

    Google Scholar 

  • Bates GW, Hasenkampf CA (1985) Culture of plant somatic hybrids following electrical fusion. Theor Appl Genet 70: 227–233

    Article  Google Scholar 

  • Bergounioux C, Perennes C, Tabaeizadeh Z, Prevot P, Gadal P (1983) Characterization and separation of Petunia hybrida and Lycopersicon esculentum protoplasts based on natural fluorescence or FITC added to label antibodies in immunoreaction with the protoplast membrane. Experientia Suppl 45: 262–263

    Google Scholar 

  • Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12: 8711–8721

    Article  PubMed  CAS  Google Scholar 

  • Bonnett HT, Glimelius K (1983) Somatic hybridization in Nicotiana: behavior of organelles after fusion of protoplasts from male-fertile and male-sterile cultivars. Theor Appl Genet 65: 213–217

    Article  Google Scholar 

  • Brown S (1984) Analysis and sorting of plant material by flow cytometry. Physiol Veg 22: 341–349

    Google Scholar 

  • Brown S, Renaudin J-P, Prevot C, Guern J (1984) Flow cytometry and sorting of plant protoplasts: technical problems and physiological results from a study of pH and alkaloids in Catharanthus roseus. Physiol Veg 22: 542–554

    Google Scholar 

  • Brown S, Jullien M, Coutos-Thevenot P, Muller P, Renaudin J-P (1987) Present developments of flow cytometry in plant biology. Biol Cell 58: 173–178

    Google Scholar 

  • Conia J, Bergounioux C, Perennes C, Muller P, Brown S, Gadal P (1987) Flow cytometric analysis and sorting of plant chromosomes from Petunia hybrida protoplasts. Cytometry 8: 500–508

    Article  PubMed  CAS  Google Scholar 

  • Davey MR, Kumar A (1983) Higher plant protoplasts retrospect and prospect. Int Rev Cytol Suppl 16: 219–299

    CAS  Google Scholar 

  • DeLaat AMM, Blaas J (1984) Flow cytometric characterization and sorting of plant chromosomes. Theor Appl Genet 67: 463–467

    Google Scholar 

  • DeLaat AMM, Shell JHN (1986) The integrity of metaphase chromosomes of Haplopappus gracilis ( Nutt.) Gray isolated by flow cytometry. Plant Sci 47: 145–151

    Article  CAS  Google Scholar 

  • Esau K (1965) Plant anatomy, 2nd edn. John Wiley & Sons, New York

    Google Scholar 

  • Evans DA, Bravo JE, Gleba YY (1983) Somatic hybridization: fusion methods, recovery of hybrids and genetic analysis. Int Rev Cytol Suppl 16: 143–159

    Google Scholar 

  • Fitter MS, Norman PM, Hahn MG, Wingate VPM, Lamb CJ (1987) Identification of somatic hybrids in plant protoplast fusions with monoclonal antibodies to plasma-membrane antigens. Planta 170: 49–54

    Article  Google Scholar 

  • Frye LD, Edidin M (1970) The rapid intermixing of cell surface antigens after formation of mouse-human heterokaryons. J Cell Sci 7: 319–335

    PubMed  CAS  Google Scholar 

  • Galbraith DW (1981) Identification and sorting of plant heterokaryons. US Dep Commerce Patent 4: 300–310

    Google Scholar 

  • Galbraith DW ( 1984a) Selection of somatic hybrid cells by fluorescence-activated cell sorting. In: Vasil IK (ed) Cell culture and somatic cell genetics of plants. Academic Press, New York London Orlando, pp 433–447

    Google Scholar 

  • Galbraith DW (1984b) Flow cytometric analysis of the cell cycle. In: Vasil IK (ed) Cell culture and somatic cell genetics of plants. Academic Press, New York London Orlando, pp 765–777

    Google Scholar 

  • Galbraith DW, Galbraith JEC (1976) A method for identification of fusion of plant protoplasts derived from tissue cultures. Z Pflanzenphysiol 93: 149–158

    Google Scholar 

  • Galbraith DW, Harkins KR (1982) Cell sorting as a means for isolating somatic hybrids. In: Fujiwara A (ed) Plant tissue culture 1982. Maruzen, Tokyo, pp 617–618

    Google Scholar 

  • Galbraith DW, Mauch TJ (1980) Identification of fusion of plant protoplasts. II. Conditions for the reproducible fluorescence labelling of protoplasts derived from mesophyll tissue. Z Pflanzenphysiol 98: 129–140

    Google Scholar 

  • Galbraith DW, Afonso CL, Harkins KR (1984) Flow sorting and culture of protoplasts: Conditions for high-frequency recovery, growth and morphogenesis from sorted protoplasts of suspension cultures of Nicotiana. Plant Cell Rep 3: 151–155

    Article  Google Scholar 

  • Galbraith DW, Harkins KR, Jefferson RA (1988) Flow cytometric characterization of the chlorophyll contents and size distributions of plant protoplasts. Cytometry 9: 75–83

    Article  PubMed  CAS  Google Scholar 

  • Galbraith DW, Harkins KR, Maddox JM, Ayres NM, Sharma DP, Firoozabady E (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 220: 1049–1051

    Article  PubMed  CAS  Google Scholar 

  • Gleba Y, Kolesnik NN, Meshkene IV, Cherep NN, Parokonny AS (1984) Transmission genetics of the somatic hybridization process in Nicotiana. I. Hybrids and cybrids among the regenerates from cloned protoplast fusion products. Theor Appl Genet 69: 121–128

    Google Scholar 

  • Gleba Y, Komarnitsky IK, Kolesnik NN, Meshkene I, Martyn GI (1985) Transmission genetics of the somatic hybridization process in Nicotiana. II. Plastome heterozygotes. Mol Gen Genet 198: 476–481

    Article  CAS  Google Scholar 

  • Glimelius K (1987) Enrichment of somatic hybrids with flow cytometry. Int Assoc Plant Tissue Cult Newslett 52: 2–10

    Google Scholar 

  • Glimelius K, Djupsjobacka M, Fellner-Feldegg H (1986) Selection and enrichment of plant protoplasts heterokaryons of Brassicaceae by flow sorting. Plant Sci 45: 133–141

    Article  CAS  Google Scholar 

  • Harkins KR, Galbraith DW (1984) Flow sorting and culture of plant protoplasts. Physiol Plant 60: 43–52

    Article  Google Scholar 

  • Harkins KR, Galbraith DW (1987) Factors governing the flow cytometric analysis and sorting of large biological particles. Cytometry 8: 60–70

    Article  PubMed  CAS  Google Scholar 

  • Haugland R (1985) Handbook of fluorescent probes and research chemicals. Molecular Probes, Junction City, Oregon

    Google Scholar 

  • Kanchanapoom K, Brightman AO, Grimes HD, Boss WF (1985) A novel method for monitoring protoplast fusion. Protoplasma 124: 65–70

    Article  Google Scholar 

  • Melamed MR, Mullaney PF, Mendelsohn ML (1979) Flow cytometry and sorting. John Wiley & Sons, New York

    Google Scholar 

  • Meyer DJ, Afonso CL, Harkins KR, Galbraith DW (1988) Characterization of plant plasma membrane antigens. In: Leaver CJ, Sze H (eds) Plant membranes: structures, function, biogenesis. UCLA Symp Molecular and cellular biology, New Ser, vol 63. Liss, New York, pp 123–140

    Google Scholar 

  • Muirhead KA, Horan PK, Poste G (1985) Flow cytometry: present and future. Biotechnology 3: 337–356

    Article  CAS  Google Scholar 

  • Pauls KP, Chuong PV (1987) Flow cytometric identification of Brassica napus protoplast fusion products. Can J Bot 65: 834–838

    Article  Google Scholar 

  • Petit PX, Conia SC, Brown SC, Bergounioux C (1986) Cytométrie en flux et biotechnologies végétales. Un avenir prometteur. Biofutur 51: 128–139

    Google Scholar 

  • Redenbaugh K, Ruzin S, Bartholomew J, Bassham JA (1982) Characterization and separation of plant protoplasts via flow cytometry and cell sorting. Z Pflanzenphysiol 107: 65–80

    Google Scholar 

  • Rotman BR, Papermaster RO (1966) Membrane properties of living cells as studied by enzymatic hydrolysis of fluorogenic esters. Proc Natl Acad Sci USA 55: 134–141

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Plant Sciences, University of Arizona, Tucson, Arizona, 85721, USA

    D. W. Galbraith

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  1. D. W. Galbraith
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Editors and Affiliations

  1. A-137 New Friends Colony, 110065, New Delhi, India

    Y. P. S. Bajaj

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© 1989 Springer-Verlag Berlin Heidelberg

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Galbraith, D.W. (1989). Flow Cytometric Analysis and Sorting of Somatic Hybrid and Transformed Protoplasts. In: Bajaj, Y.P.S. (eds) Plant Protoplasts and Genetic Engineering II. Biotechnology in Agriculture and Forestry, vol 9. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-74454-9_18

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  • DOI: https://doi.org/10.1007/978-3-642-74454-9_18

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-74456-3

  • Online ISBN: 978-3-642-74454-9

  • eBook Packages: Springer Book Archive

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