ESTCP Project #1011, Rhizosphere
Final Report
Appendix B. Plate Counts
Media for Plate Counts
Bacteria: 0.1x Tryptic Soy Agar (TSA) Medium
3.0 g Tryptic soy broth (Difco #0370-17-3)
15.0 g granulated agar
(or TSA (Difco #0369-17-6) combined TSB and agar in one product)
1.0 L distilled water
0.1 g cycloheximide (Sigma # C7698) in 1.0 mL methanol (added to medium after
autoclaving)
Autoclave @ 121C for 15 min. After cooling to approximately 45C, add
cycloheximide.
Reference: Zuberer, D.A. 1994. Recovery and enumeration of viable bacteria. p. 119-
144. In R.W. Weaver (ed.). Methods of soil analysis. Part 2.
Microbiological and biochemical properties. SSSA Book Ser. 5. SSSA,
Madison, WI.
Fungi: Martin's Medium
10.0 g glucose (dextrose)
5.0 g peptone
0.50 g KH2PO4
0.50 g K2HPO4
0.5 g MgSO47H2O
33 mg (3.3 mL) Rose Bengal*
15.0 g granulated agar
1.0 L distilled water
30.0 mg streptomycin sulfate (Sigma #S6501) (added after autoclaving)
Autoclave @ 121C for 15 min. After cooling to approximately 45C, add
streptomycin sulfate1.
*Rose Bengal, Dissolve 1.0 g Rose Bengal in 100.0 mL deionized water.
Reference: Parkinson, D. 1994. Filamentous fungi. p. 329-350. In R.W. Weaver (ed.).
Methods of soil analysis. Part 2. Microbiological and biochemical
properties. SSSA Book Ser. 5. SSSA, Madison, WI.
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