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Top : Microbiology : Hydrolytic Activity of Bacterial Supernatants for Fungal Suppression
Hydrolytic Activity of Bacterial Supernatants for Fungal Suppression | |
| Author: Shweta Bajoria, Rakhi Agrawal, Dr. R.P.Pareek, Bir | |
| Date Added: Mon Feb 02 2009 | |
| Date Modified: Mon Feb 02 2009 | |
| 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." | |
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 | |