- Can any body suggest methods to revive old Phytophthora cultures(more than 8 years old) stored in distilled water. The cultures were grown in carrot agar and the mycelial plugs are stored in vials containing distilled water.
Hi, You can culture it on PDA media directly from the stored culture, take 1 ml from the liquid stored culture and put it on PDA media, incubate at 25C for 5-7 day.
Hello, tomato agar and soybean agar were found to be comparable to or in some cases better than the popular V8 vegetable juice agar in supporting linear growth of Phytophthora. You can use PDA also as mentioned above.
I store my Phytophthora and Pythium cultures on hempseed in sterile water, and I've had them survive for at least 5 years. However, my cultures are of species that have oospores or chlamydospores. I think strictly mycelial cultures will not survive very long this way. Recently I was given a collection of Phytophthora on PDA slants that had been stored for several years and were not looking terribly good; I prepared vials of autoclaved hempseed in sterile water aa well as V-8 plates and was surprised to discover that if I added plugs to the hempseed vials I got a better recovery than on V-8. The only trick to hempseed is a) they need to be legal where you are, (if they aren't use whole rye-berries ) b) you should use sandpaper or a file to crack open the hempseed before adding it to the vial so that, upon autoclaving, the seed bursts and the endosperm is available. It will generally split open with autoclaving, but not always. I use three seeds per vial on the assumption that at least one will burst open.
A question for which I have no answer is what to do with Phytophthoras with oospores that don't readily germinate. They might be alive in your cultures but not geminating. My lab has snails that would eat the carrot agar and excrete the oospores, which might possibly then germinate on selective media. I hesitate to advise you to get snails.
Hi, Honestly, keeping alive mycelial culture of Phytophthora is difficult (or impossible) for several years. But maybe you can try your chance by putting mycelial plugs in the ryeB broth or on ryeB agar media. On PDA medium, they usually grow very slow in comparison with ryeB/A agar media. Good luck.
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