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Chemically defined medium for cultivation of several epiphytic and phytopathogenic spiroplasmas

I M Lee1, R E Davis

  • 1Department of Botany, University of Maryland, College Park, Maryland 20742 and Plant Virology Laboratory, Agricultural Research Service, United States Department of Agriculture, Beltsville, Maryland 20705.

Applied and Environmental Microbiology
|December 1, 1983
PubMed

Insights

A new chemically defined medium, LD82, supports the in vitro cultivation of multiple spiroplasma species. This medium facilitates good growth and colony formation for both epiphytic and phytopathogenic spiroplasmas.

Area of Science:

  • Microbiology
  • Cell Biology
  • Biotechnology

Background:

  • Spiroplasmas are helical, wall-less bacteria with diverse ecological roles.
  • In vitro cultivation of spiroplasmas is crucial for studying their biology and pathogenicity.
  • Existing culture media often lack precise chemical definition, hindering reproducible research.

Purpose of the Study:

  • To formulate and evaluate a chemically defined medium (LD82) for spiroplasma in vitro cultivation.
  • To assess the growth-supporting capabilities of LD82 for various spiroplasma strains.
  • To investigate the preliminary nutritional requirements of spiroplasmas in the defined medium.

Main Methods:

  • Formulation of a novel chemically defined medium, designated LD82.
  • Cultivation of six spiroplasma strains (four epiphytic/insect-pathogenic, two phytopathogenic) in LD82.
  • Quantification of spiroplasma titers (CFU/ml) and assessment of colony morphology on LD82 agar.
  • Preliminary nutritional studies, including the effect of phospholipids.

Main Results:

  • Medium LD82 supported good growth for all six tested spiroplasma strains, reaching titers of 2.0 x 10^6 to 6.0 x 10^7 CFU/ml.
  • All spiroplasma strains readily formed diffuse colonies on LD82 agar, though colony morphology varied.
  • Phospholipids showed a slight growth-enhancing effect on epiphytic/insect-pathogenic strains and were essential for phytopathogenic Spiroplasma citri.

Conclusions:

  • The chemically defined medium LD82 is effective for the in vitro cultivation of diverse spiroplasma species.
  • LD82 provides a reliable platform for spiroplasma research, enabling consistent growth and colony formation.
  • Phospholipids play a significant role in the nutritional requirements of certain spiroplasma groups, particularly Spiroplasma citri.

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