Be 2.3 Pseudomonas syringae pv syringae 2017-05-08T14:10:07+00:00

National Seed Health System

[printfriendly]

Be 2.3      Pseudomonas syringae pv syringae

VERSION: 1.0
DATE: 12/2012
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:

1. 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.

2. Stir each suspension thoroughly with a sterile glass rod and then remove a 20 ml sample.

3. Make three serial tenfold dilutions in SR and plate 0.1 ml of each dilution (three duplicates of each).

4. 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.