Dear Bimla Kaira

Pls. find the attached files

https://research.cip.cgiar.org/typo3/web/fileadmin/icmtoolbox/ICM_Toolbox/Files/Manual_draft1.pdf

http://agrifoodscience.com/index.php/TURJAF/article/viewFile/325/219

Hoping this will be helpful            

Regards

Prof. Houda Kawas

I have several suggestions that you could try:

1) Plate serial cultures on Rye Agar with either PARP or RVN (antibiotics), each time taking the smallest possible piece of inoculum from the leading edge of the growing mycelium.

2) Use a very dilute suspension of zoospores in sterile water to inoculate Rye Agar (again, PARP or RVN) with very small droplets. If one of the droplets is diluted to the point of only a single spore, you may achieve isolation.

3) Inoculate surface sterilized leaflets with single droplets of inoculum, incubate in sterile humid conditions until symptoms appear, and then collect mycelium from lesions outside the inoculation site.

4) Gently filter a zoospore suspension using filters of decreasing pore size. The fungal mycelium appears larger than P. infestans zoospores, so this may work.

5) Use the zoospores' motility to separate them from the (potentially) less mobile fungi. Gently centrifuge the inoculum, give the zoospores a little time to move around and then carefully decant only the upper portion of the suspension. Repeat, if necessary.

I think I may like #5 the best, if the fungal spores are non-motile.

Best of luck with your research!

 Dear Bimla Kaira,

I thing your culture  contaminated with  Fusarium sp you must purified the fungus Phytophthora within 2-3 days because its  grow faster  than  Fusarium sp. So if you delayed the two fungus will grow together and it will be so difficult to get pure culture of  Phytophthora sp.

I normally isolated Phytophthora infestans by plating sporangia or small pieces of infected tissue onto antibiotic rye agar including the anti-fungal antibiotic pimaricin (natamycin) to suppress fungal contamination as well as an antibacterial antibiotic (rifamycin).  The media are described in the CIP Manual referenced above.

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Hi Dr.Bimla

Good Day

I agree with Dr.Neran , phytophthora contamination with Fusarium

You use bait for Phytophthora because Fusarium is growth quickly

Therefore, you can use the bait lupin seeds or cocoa pods

Thank you

Sincerely