Hi,
I am conducting an experiment for reisolate Verticillium dahliae from cacao. I am thinking to grind my sample in phosphate buffer and PVP (15 g/l). I am adding PVP because my samples are quite rich in polyphenols that could impact the reisolation success. I want to use a buffer at a molarity of 1M or 2 M because they are also quite rich in polysaccharides and so they are viscous. Does this could impact the reisolation success?
Is it not possible to isolate just by crushing the sample in distilled water? Why do it needs buffer?
If the fungus is endophytically growing in the plants in presence of all those polyphenols, then how can polyphenols create problem is isolation of the fungus?
I'm going to sound 120 years old, but the way we old-timers isolate Verticillium is to plate a piece of tissue containing vascular elements so the xylem is perpendicular to the surface of the plate. The Verticillium will grow out of the xylem and it is possible to use a dissecting needle to touch the emerging conidiophores and collect spores for isolation.
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