Laboratory Maintenance of Streptomyces Species

Micah D. Shepherd1, Madan K. Kharel1, Mary A. Bosserman1, Jürgen Rohr1

1 College of Pharmacy, University of Kentucky, Lexington, Kentucky
Publication Name:  Current Protocols in Microbiology
Unit Number:  Unit 10E.1
DOI:  10.1002/9780471729259.mc10e01s18
Online Posting Date:  August, 2010
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Abstract

This unit includes general protocols for the laboratory maintenance of Streptomyces species, including growth in liquid media, growth on solid agar, and short‐ and long‐term storage. Considerations for the handling of Streptomyces species and the morphology of the bacteria are also reviewed. Curr. Protoc. Microbiol. 18:10E.1.1‐10E.1.8. © 2010 by John Wiley & Sons, Inc.

Keywords: Streptomyces; media; strain storage; spore; mycelium

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Growth of Streptomyces in Liquid Media
  • Basic Protocol 2: Growth of Streptomyces on Solid Media
  • Basic Protocol 3: Storage of Streptomyces on Solid Media
  • Basic Protocol 4: Storage of Streptomyces mycelia as Glycerol Stocks
  • Basic Protocol 5: Storage of Streptomyces Spores as Glycerol Stocks
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Growth of Streptomyces in Liquid Media

  Materials
  • Sterile liquid medium [e.g., Tryptic soy‐broth (TSB), R2YE, and YEME media; see reciperecipes]
  • Agar plate of a Streptomyces species
  • 250‐ml Erlenmeyer baffled flasks
  • Cotton gauze
  • Aluminum foil
  • Autoclave
  • Biological safety cabinet
  • Sterile toothpicks
  • Sterile disposable inoculation loops
  • 28°C shaking incubator

Basic Protocol 2: Growth of Streptomyces on Solid Media

  Materials
  • Fully grown liquid Streptomyces culture ( protocol 1)
  • Agar medium (see reciperecipes)
  • Petri dishes
  • Sterile plastic spreader
  • Biological safety cabinet
  • 28°C incubator
  • Sterile toothpicks
  • Microscope
  • Parafilm

Basic Protocol 3: Storage of Streptomyces on Solid Media

  Materials
  • Fresh agar plate of the Streptomyces strain ( protocol 2)
  • Refrigerator (4°C)
  • Additional reagents and equipment for preparing fresh mycelia on solid medium ( protocol 2)

Basic Protocol 4: Storage of Streptomyces mycelia as Glycerol Stocks

  Materials
  • Fresh agar plate or liquid culture of the Streptomyces strain ( protocol 1 or 2, respectively)
  • Sterile water
  • 50% glycerol in water (sterilized by autoclaving)
  • Sterile 2‐ml screw‐top vials or 1.5‐ml microcentrifuge tubes (for mycelia storage)
  • 1‐ml pipet tips
  • Sterile loop or sterile toothpick
  • −80°C freezer

Basic Protocol 5: Storage of Streptomyces Spores as Glycerol Stocks

  Materials
  • Fresh sporulating agar plate (or slant) of the strain ( protocol 2)
  • Sterile water
  • 50% glycerol in water (sterilized by autoclaving)
  • Heavy‐duty aluminum foil
  • Sterile spreader or sterile loop
  • Sterile pipet
  • Sterile 50‐ml screw‐top vials (for spore collection and preparation)
  • Vortex mixer
  • Sterile cotton filter (puncture the bottom of a 50‐ml conical screw‐top vial and gently pack roughly 2 cm of cotton to the bottom of the vial; after packing the cotton, re‐attach the screw top and wrap the entire vial with aluminum foil and autoclave)
  • Centrifuge
  • Sterile 2‐ml screw‐top vials or 1.5‐ml microcentrifuge tubes (for spore storage)
  • −80°C freezer
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Figures

Videos

Literature Cited

Literature Cited
   Kieser, T., Bibb, M.J., Buttner, M.J., Chater, K.F., and Hopwood, D.A. 2000. Practical Streptomyces Genetics. The John Innes Foundation, Norwich, England.
   Miyadoh, S. 1993. Research on antibiotic screening in Japan over the last decade: A producing microorganisms approach. Actinomycetologica 9:100‐106.
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