Archived 04.03.2018 Be 2.3 – Pseudomonas syringae pv syringae
| Version | 1.0 |
|---|---|
| Date | 12/2012 [Archived 04.03.2018] |
| Pathogen | Pseudomonas syringae pv. syringae |
| Host | common bean (Phaseolus vulgaris) |
| Common Name | bacterial brown spot |
| Method | Be 2.3 Semi-selective Media (Gszczynska and Serfontein 1998) |
| Method Class | Temporary Standard (B) |
| Sample | 1 kg of seeds |
Procedure:
- Soak two samples of 1 kg seed per seed lot in SR buffer (2 ml per gram of seed) in sterile containers for 20 h at 5°C.
- Stir each suspension thoroughly with a sterile glass rod and then remove a 20 ml sample.
- Make three serial tenfold dilutions in SR and plate 0.1 ml of each dilution (three duplicates of each).
- Incubate plates at 28°C for 4–5 days and then count the number of pathogens and saprophytes.
Media recipe:
- Solution 1: proteose peptone no. 3 (Difco) (10 g), CaCl2 (0.25 g), tyrosine (1 g), agar (15 g), distilled water (500 ml);
- Solution 2: skim milk (10 g), distilled water (500 ml);
- Solution 3: Tween 80 (10 ml). Autoclave solutions separately and mix when still hot. Cool to 50°C and add the following antibiotics under sterile conditions: cephalexin 80 mg/l, cycloheximide 200 mg/l and vancomycin 10 mg/l.
References:
Goszczynska, T. and Serfontein, J. J. 1998. Milk-Tween agar, a semiselective medium for isolation and differentiation of Pseudomonas syringae pv. syringae, Pseudomonas syringae pv. phaseolicola and Xanthomonas axonopodis pv. phaseoli. Journal of Microbiological Methods. 32(1): 65-72.