Somatic hybridization in the gramineae: Pennisetum americanum (L.) K. Schum. (Pearl millet) +Panicum maximum Jacq. (Guinea grass) View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1986-06

AUTHORS

Peggy Ozias-Akins, Robert J. Ferl, Indra K. Vasil

ABSTRACT

Protoplasts from Pennisetum americanum resistant to S-2-amino-ethyl-l-cysteine (AEC) were fused with protoplasts of Panicum maximum utilizing polyethylene glycol-dimethylsulfoxide after inactivation of the Pennisetum protoplasts with 1 mM iodoacetic acid. The iodoacetate treatment prevented division of Pennisetum protoplasts; therefore, only Panicum protoplasts and heterokaryons potentially could give rise to colonies. A second level of selection was imposed by plating 3–4-week-old colonies on AEC medium. Putative somatic hybrid calli were analyzed for alcohol dehydrogenase, 6-phosphogluconate dehydrogenase, aminopeptidase, and shikimate dehydrogenase isozymes. Three somatic hybrid cell lines (lines 2, 3, and 67) were identified which showed two bands of alcohol dehydrogenase activity representing homodimers of P. maximum and P. americanum as well as a novel intermediate band of activity where Panicum-Pennisetum heterodimers would be expected. Aminopeptidase and shikimate dehydrogenase were useful for identifying presumptive hybrid calli but the isozyme patterns were additive-evidence which would not preclude the selection of chimeric callus. A more complex isozyme pattern which varied among the somatic hybrids was observed for 6-phosphogluconate dehydrogenase. In the hybrid calli, the presence of DNA sequences homologous to both P. maximum and P. americanum sequences was confirmed by hybridization of a maize ribosomal DNA probe to XbaI and EcoRI restriction fragments. Growth of hybrid lines on various concentrations of AEC was either similar to the AEC-resistant parent (hybrid line 2) or intermediate between the resistant and sensitive parents (hybrid lines 3, 67). More... »

PAGES

365-370

References to SciGraph publications

  • 1974-12. A method for high-frequency intergeneric fusion of plant protoplasts in PLANTA
  • 1974-01. Plant protoplast fusion and growth of intergeneric hybrid cells in PLANTA
  • 1984-04. Species-specific repetitive DNA used to identify interspecific somatic hybrids in PLANT CELL REPORTS
  • 1975-01. Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media in PLANTA
  • 1983-01. Detection of two different nuclear genomes in parasexual hybrids by ribosomal RNA gene analysis in THEORETICAL AND APPLIED GENETICS
  • 1980-10. The use of cytoplasmic streptomycin resistance: Chloroplast transfer from Nicotiana tabacum into Nicotiana sylvestris, and Isolation of their somatic hybrids in MOLECULAR GENETICS AND GENOMICS
  • 1977-01. Isoenzyme patterns in soybean-Nicotiana somatic hybrid cell lines in MOLECULAR GENETICS AND GENOMICS
  • 1983-10. The use of amino acid analog resistance and plant regeneration ability to select somatic hybrids between Nicotiana tabacum and N. glutinosa in MOLECULAR GENETICS AND GENOMICS
  • 1980-05. Isolation and culture of cereal protoplasts in THEORETICAL AND APPLIED GENETICS
  • 1979-05. Use of amino acid analogue-resistant cell lines for selection of Nicotiana sylvestris somatic cell hybrids in THEORETICAL AND APPLIED GENETICS
  • 1981-07. Chloroplast transfer in Nicotiana based on metabolic complementation between irradiated and iodoacetate treated protoplasts in PLANTA
  • 1984-10. High frequency of fusion induced in freely suspended protoplast mixtures by polyethylene glycol and dimethylsulfoxide at high pH in PLANT CELL REPORTS
  • 1983-09. A plant DNA minipreparation: Version II in PLANT MOLECULAR BIOLOGY REPORTER
  • 1985-04. Somatic hybridization between Lycopersicon esculentum and Lycopersicon pennellii in THEORETICAL AND APPLIED GENETICS
  • Journal

    TITLE

    Molecular Genetics and Genomics

    ISSUE

    3

    VOLUME

    203

    Author Affiliations

    Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/bf00422058

    DOI

    http://dx.doi.org/10.1007/bf00422058

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1032574803


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    38 schema:description Protoplasts from Pennisetum americanum resistant to S-2-amino-ethyl-l-cysteine (AEC) were fused with protoplasts of Panicum maximum utilizing polyethylene glycol-dimethylsulfoxide after inactivation of the Pennisetum protoplasts with 1 mM iodoacetic acid. The iodoacetate treatment prevented division of Pennisetum protoplasts; therefore, only Panicum protoplasts and heterokaryons potentially could give rise to colonies. A second level of selection was imposed by plating 3–4-week-old colonies on AEC medium. Putative somatic hybrid calli were analyzed for alcohol dehydrogenase, 6-phosphogluconate dehydrogenase, aminopeptidase, and shikimate dehydrogenase isozymes. Three somatic hybrid cell lines (lines 2, 3, and 67) were identified which showed two bands of alcohol dehydrogenase activity representing homodimers of P. maximum and P. americanum as well as a novel intermediate band of activity where Panicum-Pennisetum heterodimers would be expected. Aminopeptidase and shikimate dehydrogenase were useful for identifying presumptive hybrid calli but the isozyme patterns were additive-evidence which would not preclude the selection of chimeric callus. A more complex isozyme pattern which varied among the somatic hybrids was observed for 6-phosphogluconate dehydrogenase. In the hybrid calli, the presence of DNA sequences homologous to both P. maximum and P. americanum sequences was confirmed by hybridization of a maize ribosomal DNA probe to XbaI and EcoRI restriction fragments. Growth of hybrid lines on various concentrations of AEC was either similar to the AEC-resistant parent (hybrid line 2) or intermediate between the resistant and sensitive parents (hybrid lines 3, 67).
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