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Importance of myo-inositol, calcium, and ammonium for the viability and division of tomato (Lycopersicon esculentum) protoplasts

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Abstract

Plants from four cultivars of Lycopersicon esculentum were grown under different conditions, in controlled environment chambers. Low light intensity, long photoperiod (16 h), 25° C/17°C temperature alternance (day/night) were found to be the most convenient conditions for obtaining viable protoplasts. The use of myo-inositol as an osmoticum in the digestion medium and the adjustment of the pH to 6.5, instead of the usual 5.8, for this medium increased the yield of viable protoplasts and enhanced their stability. Under these conditions neither pretreatment (dark and cold treatments), nor preplasmolysis of leaf tissues, were required before protoplast isolation. The concentrations of ammonium nitrate, calcium chloride, myo-inositol, and sucrose were found to be critical for the success of protoplast culture. A medium containing 5 mM ammonium nitrate, 40 mM calcium chloride, 10 mg l-1 adenine sulfate, 0.5% myo-inositol and 6% sucrose gave sustained protoplast divisions. Under these conditions, plating efficiency ranged from 5% for the cultivar Lukulus to 15% for the cultivar Golden Sunrise.

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Abbreviations

BA:

benzylaminopurine

CaCl2 :

calcium chloride, 2,4,-D-2,4-dichlorophenoxyacetic acid

EDTA:

ethylene diamine tetraacetic acid

KCl:

potassium chloride

MES-2-N:

morpholino ethane sulfonic acid

MgCl2 :

magnesium chloride

NH4NO3 :

ammonium nitrate

NAA:

naphthalene acetic acid, p-protoplasts

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Authors and Affiliations

  1. Laboratoire de Biologie Cellulaire, I.N.R.A., F-78026, Versailles-Cedex, France

    Catherine Bellini, Marie-Christine Chupeau, Monica Gervais, Gérard Vastra & Yves Chupeau

  2. Société Roussel Uclaf, F-93230, Romainville, France

    Catherine Bellini, Marie-Christine Chupeau, Monica Gervais, Gérard Vastra & Yves Chupeau

Authors
  1. Catherine Bellini
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  2. Marie-Christine Chupeau
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  3. Monica Gervais
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  4. Gérard Vastra
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  5. Yves Chupeau
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Bellini, C., Chupeau, MC., Gervais, M. et al. Importance of myo-inositol, calcium, and ammonium for the viability and division of tomato (Lycopersicon esculentum) protoplasts. Plant Cell Tiss Organ Cult 23, 27–37 (1990). https://doi.org/10.1007/BF00116086

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  • DOI: https://doi.org/10.1007/BF00116086

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