In our experiments, We need lots of zoospores to infect potato plants and see their activity. How is the best way for getting zoospore?
Hi Seyed: Please find these useful links
Article An efficient method for zoospore production, infection and r...
http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.626.4797&rep=rep1&type=pdf
http://arizona.openrepository.com/arizona/bitstream/10150/347975/1/AZU_TD_BOX176_E9791_1976_310.pdf
Hello, Thank you Oadi for the links. Actually, I am looking for a handy protocol for P. infestanse?
I have read several papers for Zoospore production and could not find an applicable for P. infestanse.
Hi Seyed
The way we do it in my lab is
> first, to produce and collect sporangia from leaves ( detached leaves work OK): just pick leaflets from 3-4 week old potato leaves, deposit each leaflet on the lid of an inverted Petri dish into which you had poured water agar (the idea is that the dish works as a moist chamber, without the leaflet touching the agar to avoit it to stick : so pour the agar into the dish, let it cool, then invert the dish so the agar is on top and the lid on the bottom, and then palce the leaflet on the lid before closing the dish again), put a drop of sporangia suspension , wathever the concentration, on the lower side of leaflets, and incubate at toom temperature for 4-7 days depending on temperature and isolate. Once you get sporulating lesions, you can collect the sporangia produced by picking the leaflet from the leaf, putting it into a screw acp tube with 5-10 ml sterile water, then, shake gently before removing the leaflet and keeping the spore suspension.
> then to realease the zoospores, cool; the suspension to about 4°C for 2-4 h. This elicits zooposre relase from sporangia. The ideal way would be to do that under agitation so the zoospores don't settle and encyst.
You can use the cooling procedure also with sporangia prtoduced from axenic cultures on nutrient agar ( rye agar, pea agar and V8 agar work usually best), although you might loose some pathogenicity compared to sporangia produced on leaves.
Hello Didier,
Thank you very much for your complete explanations....:)
Hope to see you again.
Mahyar
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