National Seed Health System

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ARCHIVED 02.21.2025     Be 4.2 – Curtobacterium flaccumfaciens pv. flaccumfaciens

VERSION: 1.0
DATE: 12/2012  [ARCHIVED 02.21.2025]
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.