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Culturing shoot-tip clumps of pearl millet [Pennisetum glaucum (L.) R. Br.] and optimal microprojectile bombardment parameters for transient expression

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Abstract

Microprojectile bombardment and transient expression of the reporter gene, β-glucuronidase (GUS) in a novel target tissue, multiple-shoot-tip clumps of pearl millet [Pennisetum glaucum (L.) R. Br.] is reported here. The multiple-shoot-tip clumps were developed in vitro from shoot-apices of seedlings. Using this method, the apical meristems along with the germline cells were easily exposed for bombardment without loss of viability. Further growth of the multiple-shoot-tip clumps was not substantially affected by microprojectile bombardment. Transient expression of β-glucuronidase gene was detected in the form of blue transformed cell sectors in the bombarded tissue by an in situ enzyme assay. The blue sectors were used as convenient criteria to study several factors affecting gene transfer efficiency. Optimal conditions for efficient transient expression of the GUS gene have been defined to aid future strategies of genetic engineering in pearl millet with agronomically important genes.

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Authors and Affiliations

  1. Biotechnology Laboratory, Department of Botany, Osmania University, Hyderabad, 500007, India

    Prathibha Devi

  2. Genetic Engineering Laboratory, Department of Crop and Soil Sciences, Michigan State University, East Lansing, MI, 48824, U.S.A

    Mariam Sticklen

Authors
  1. Prathibha Devi
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  2. Mariam Sticklen
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Devi, P., Sticklen, M. Culturing shoot-tip clumps of pearl millet [Pennisetum glaucum (L.) R. Br.] and optimal microprojectile bombardment parameters for transient expression. Euphytica 125, 45–50 (2002). https://doi.org/10.1023/A:1015733715791

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  • DOI: https://doi.org/10.1023/A:1015733715791

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