Tween medium
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Contents |
Purpose
This medium was developed for isolations of Xanthomonas campestris pv. vesicatoria from leaves. Variations have been developed for strains less tolerant of boric acid, and for isolations of X. campestris pv. vesicatoria from soils.
Ingredients
| Ingredient | 1 L | 500 ml |
|---|---|---|
| Part 1 | ||
| Distilled water | 1000 ml | 500 ml |
| Peptone | 10 g | 5 g |
| KBr | 10 g | 5 g |
| CaCl2 | 0.25 g | 0.125 g |
| Boric acid | 0.3 g | 0.15 g |
| Part 2 | ||
| Agar | 15 g | 7.5 g |
| Part 3 | ||
| Cycloheximide | 0.05 g | 0.025 g |
| Cephalexin | 0.0.65 g | 0.0325 g |
| 5-Fluorouracil | 0.012 g | 0.006 g |
| Tobramycin | 0.0004 g | 0.0002 g |
Instructions
- Adjust pH of ingredients from part 1 to 7.4 with NaOH, then add ingredient from part 2.
- Autoclave the medium in a flask with a stir bar for 15 min.
- Also autoclave 10 ml of Tween 80 in a separate container.
- Add the Tween to the hot medium immediately after removing the medium from the autoclave.
- Shaking the flask after adding the Tween produces bubbles, so a magnetic stir plate is recommended.
- The medium will appear cloudy after the tween is added but will clear as it cools. Cool to about 55 °C before adding antibiotics.
- Aseptically add aqueous solutions of the antibiotics in part 3.
Notes
References
- McGuire, R. G., Jones, J. B., and Sasser, M. 1986. Tween media for semi-selective isolation of Xanthomonas campestris pv. vesicatoria from soil and plant material. Plant Disease 70: 887-891.
Contributed by
From the Culture Media for Plant Pathogenic Fungi and Bacteria, University of Massachusetts: Contributed by Robert L. Wick
