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Complementary methodologies to identify specific agrobacterium strains.

H Bouzar1, L W Moore

  • 1Department of Botany and Plant Pathology, Oregon State University, Corvallis, Oregon 97331.

Applied and Environmental Microbiology
|November 1, 1987
PubMed
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Serological techniques and DNA restriction enzyme analysis effectively differentiate Agrobacterium strains. These complementary methods accurately identified specific strains, even after field inoculation.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Plant Pathology

Background:

  • Accurate identification of Agrobacterium strains is crucial for agricultural applications and research.
  • Existing methods may lack the specificity or robustness required for reliable strain differentiation.

Purpose of the Study:

  • To develop and validate complementary serological and DNA-based methods for differentiating Agrobacterium strains.
  • To assess the reliability of these methods for identifying specific strains, including those used in field applications.

Main Methods:

  • Serological analysis using immunodiffusion and immunofluorescence with polyclonal antisera against Agrobacterium ribosomal preparations.
  • DNA analysis involving restriction enzyme (EcoRI) digestion of total DNA to generate cleavage patterns.

Related Experiment Videos

  • Testing of 45 wild-type and plasmid-cured Agrobacterium strains.

    • Main Results:

    • Immunodiffusion yielded single, strain-specific precipitin lines, while immunofluorescence showed some cross-reactions with reduced fluorescence intensity.
    • Restriction enzyme digestion of DNA revealed 34 distinct cleavage patterns, correlating with chromosomal background and serological reactions.
    • Plasmid presence or absence did not affect serological reactions or DNA profiles.

    Conclusions:

    • Combined serological and DNA restriction profiling provides a robust and complementary approach for Agrobacterium strain identification.
    • These validated methods successfully identified Agrobacterium radiobacter K84 strains recovered from field-inoculated trees.