I am currently doing my Honors project, have Fusarium fungi culture growing on PDA plates. I need to make the mycelium growth from the PDA plates into a liquid suspension, so that l can inoculate bell pepper flower later. If anybody has a protocol or know how l should proceed please let me know. Thank you all.
Fatna Diriye
Hi Fatna,
I have worked on Fusarium oxysprom on tomate, and to inoculate my plants I prepare a suspension of 106 conidia/ml in minimal liquid medium (MLM).
You can find in attachment my article.
If you have further questions, don't hesitate.
Dr.HAMINI-KADAR N.
- transfer your fusarium from PDA into PDB (potato dextrose broth) and then incubate in the incubator with saking 120 rpm for 7-14 days. after that, filter your culture suspension using sterile filter with pore diameter 0.45 um (mikrometer) to separate the spore with mycelium. your spore is on the suspension and you can count how much your spore concentration and apply to the plant
Hi Fatna!
I worked in a similar topic, inoculating grapes with Botrytis. Here I show you the protocol I followed:
1. With an sterile loop, scrape some of the mycelium from the PDA plate. It is better to do so in the outer part of the culture because the culture is younger in that part and actively growing.
2. Place the mycelium in a tube with 5 ml of sterile deionized water with 0.1% Tween 80 or 85. Stirre using a vortex to homoemize the fungal suspension.
3. Place in a ultrasound bath for about 5 min to dettach spores from mycelium.
4. Using a cell counter measure the spores concentration and adjust your necessary concentration if required. I was using 10^4 espores/ml for inoculating.
You can use the spores suspension for inoculating your samples.
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