Hi. I'm trying to check the resistance of my tomato plant transformants against fungus like Botrytis cinerea. However, I barely work with fungus before. I have read several methods from published papers, but the protocol is not in detail. It would be very generous if anyone could help me with fungal inoculum preparation and the artificial infection of the fungus onto the host plants.
There are several methods that you can use to confirm the plant resistance.
In my case, I followed the protocols in the attached files (Widiastuti et al, 2011).
Please take a look at the paper, especially about "Growth and maintenance of fungi and plants" and the inoculation test (spray and drop mycelium methods).
There are some additional points as follows;
- You have to ensure that your B.cinerea is isolated from "single spore", generally known as single spore isolation. If you buy the isolates, you can ignore this point.
- In this paper, she used two methods of inoculation test, you can choose one of them. I prefer to apply the spray methods to do the inoculation test and use plants without cutting the leaves (spray directly onto the plants). The preparation of spore suspension is similar to that explained above.
- After inoculation, put the plants in the big box, cover it with a clear plastic bag (to maintain high humidity), and leave it for 24 h at 23C-25C. After 24 h, open the cover (plastic) and leave it until 7 days post-inoculation. Don't forget to give water to the plant every day (bottom watering plants).
- Seven days post-inoculation, we can check the symptoms on the first leaves of the plants (GMO and NT) using the scoring or image analysis methods.
Good luck!
+Induction+of+disease+resistance+against+Botrytis+cinerea+by+heat+shock+treatment+in+melon+%28Cucumis+melo+L.%29.pdf
Thank you all! Nawab Khan Otar Shainidze Kouadri Mohamed El Amine Agung Dian Kharisma
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