Abstract: As the fungal growth suppression by biocontrol agents (BCA) in solid media using dual plate assay has some issues regarding nutrient limitation. A protocol for fungal suppression in liquid medium containing bacterial supernatant in different ratios is being designed in the name of "Hydrolytic Activity of bacterial supernatants for fungal suppression."
Date Added: 2009-02-02
Date Modified: 2009-02-02
Procedure -
Grow bacterial cells in 100 ml Waksman broth at 110 rpm at 28°C till the O.D reaches 0.590 at 650 nm.
-
Centrifuge the suspension and discard the pellet.
- Take the supernatant in a separately autoclaved flask with Waksman broth in different ratios (1:1, 1:2, and 1:3).
- Inoculate the dual liquid medium with a fungal (plant pathogen) block of 5mm diameter.
- Keep the flask at 110rpm at 28°C for 4 days.
- Keep the positive control having fungal culture in Waksman broth.
- Observe the medium for fungal mycelium comparing with control.
- Instead nutrients were supplied to the fungus in the medium; bacterial supernatant will inhibit the fungal growth, if having some antifungal metabolites.
Reagents |
Waksman broth
| | Constituents | (g/lit) | |
Protease peptone
|
5.0
| |
Glucose
|
10.0
| |
Beef Extract
|
3.0
| |
NaCl
|
5.0
| |
pH 6.8
|
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