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An efficient particle bombardment system for the genetic transformation of asparagus (Asparagus officinalis L.)

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Abstract

The microprojectile bombardment method was used to transfer DNA into embryogenic callus of asparagus (Asparagus officcinalis L.) and to produce stably transformed asparagus plants. Embryogenic callus, derived from UC 157 and UC72 asparagus cultivars, was bombarded with tungsten particles coated with plasmid DNA that contained genes encoding hygromycin phosphotransferase, phosphinothricin acetyl transferase and β-glucuronidase. Putatively transformed calli were identified from the bombarded tissue after 4 months selection on 25 mg/L hygromycin B plus 4 mg/L phosphinothricin (PPT). By selecting embryogenic callus on hygromycin plus PPT the overall transformation and selection efficiencies were substantially improved over selection with hygromycin or PPT alone, where no transgenic clones were recovered. The transgenic nature of the selected material was demonstrated by GUS histochemical assays and Southern blot hybridization analysis. Transgenic asparagus plants were found to withstand the prescribed levels of the PPT-based herbicide BASTATM for weed control.

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Abbreviations

GUS:

β -glucuronidase

HPT:

hygromycin phosphotransferase

bar :

phosphinothricin acetyl transferase gene

PPT:

phosphophinothricin

NAA:

naphthalene acetic acid

2iP:

2-isopentenyl adenine

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Authors and Affiliations

  1. Departamento de Ingeniería Genética de Plantas, Centro De Investigacion y de Estudios Avanzados del IPN, Unidad Irapuato, Apartado Postal 629, 36500 Irapuato, Guanajuato, Mexico

    Jose Luis Cabrera-Ponce, Liliana López, Nacyra Assad-Garcia, Consuelo Medina-Arevalo, Ana Maria Bailey & Luis Herrera-Estrella

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  1. Jose Luis Cabrera-Ponce
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  2. Liliana López
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  3. Nacyra Assad-Garcia
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  4. Consuelo Medina-Arevalo
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  5. Ana Maria Bailey
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  6. Luis Herrera-Estrella
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Communicated by I. K. Vasil

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Cabrera-Ponce, J.L., López, L., Assad-Garcia, N. et al. An efficient particle bombardment system for the genetic transformation of asparagus (Asparagus officinalis L.). Plant Cell Reports 16, 255–260 (1997). https://doi.org/10.1007/BF01088276

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  • DOI: https://doi.org/10.1007/BF01088276

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