I am trying to quantify Phytopthora propagules from soil sample on PARPH media, but I am having difficulty keeping other fungi from growing on the plate. Can someone suggest as to how I can stop any other fungi growing on the plate? If anyone can share picture or info of the cultural characteristics of Phytopthora on culture media
You can stop growing fungi by using some fungicides like thiram or benzamidazol. These fungicides did not affect phytopthora. Another way is to use selective media for phytopthora
I had very good experience using PARPNH, but it worked when I isolated Ph. from wood samples and I didn't experience fungi or bacteria contaminations.
You can try to use an apple bait: make a hole into an apple, fill it with your soil samples and cover the apple with a plastic film. After several days, transfer a portion of colonized apple into a petri dish containing PDA, CMA, V8A or whatever medium good for Ph.
With this method, I had excellent results.
Good luck.
You have right Andrea, I had used this method befor for isolation of Phytopthora megasperma from soil
Do you think apple bait method can be used to quantify. As I am also trying to count propagules in different sample of soil. Basically I am allowing soil of known volume to soak for 48 to 72 hr and than dilute about 10cm3 of soil in WA and using 1ml of this slurry to plate.
How can apple bait method be applied to quantify?
I have used leaf bait method to isolate and it was very effective too to isolate Phytopthora, but it cannot be used to quantify.
I am thankful to you guys to take time to respond to my questions, it was real help.
I think the apple bait technique can help you just to isolate phytophthora but for use it for quantification i am not sure. The good and accurate way for quantifying phytophthora it could be real time pcr by developing a specific quantification based on target gene or protein!!!
Quantifying using real time pcr is another component of my project. But I have to successfully count propagules on culture as well.
Please see these links can help you to do for what are you looking???
http://www.apsnet.org/publications/phytopathology/backissues/Documents/1995Articles/Phyto85n10_1057.pdf
http://onlinelibrary.wiley.com/doi/10.1046/j.1365-3059.2000.00478.x/pdf
http://www.eurogentec.com/egt/files/ippolitoejpp2004.pdf
Good luck!!
- Badges
- Science topic
- Similar topics
- Biological Science
- Biota
- Eukaryota
- Fungi