Be 4.2 Curtobacterium flaccumfaciens pv. flaccumfaciens 2018-04-10T14:25:42+00:00

National Seed Health System

Be 4.2 Curtobacterium flaccumfaciens pv. flaccumfaciens

VERSION: 1.0
DATE: 12/2012
PATHOGEN: Curtobacterium flaccumfaciens pv. flaccumfaciens (syn: Corynebacterium flaccumfaciens pv flaccumfaciens)
HOST: common bean (Phaseolus vulgaris)
COMMON NAME: bacterial wilt
METHOD: Be 4.2 Selective Media Assay (Maringoni et al., 2006)
METHOD CLASS: TEMPORARY STANDARD (B)
SAMPLE: 200g seeds

PROCEDURE:

1. Soak 200g samples of bean seeds, in 600 mL sterilized distilled water for 24 h at 5°C.

2. Stir the liquid and then use for seeding by streaking with a wire loop onto the surface of the semi-selective culture medium.

3. Incubate at 28 – 30°C for 96 to 120 hours and compare the colony characteristics to pure standard isolate (they used Feij-2634).

CFFSM Recipe:

Peptone 5g 
meat extract 3g
sucrose 5g 
agar 15g 
skim milk powder* 5g 
Congo red* 0.05g 
chlorothalonil* 0.01g
thiophanate methyl* 0.01g 
nalidixic acid* 0.01g 
nitrofurantoin* 0.01g
oxacillin* 0.001g 
sodium azide* 0.001g
distilled water 1L

*added after autoclaving the basal medium

REFERENCES:

Maringoni, A. C. and Camara, R. C. 2006. Curtobacterium flaccumfaciens pv. flaccumfaciens detection in bean seeds using a semi-selective culture medium. Brazilian Journal of Microbiology. 37: 451-455.