We have recently had trouble with sub-culturing some fungal isolates that were obtained from soil and different avocado tissues (leaves, roots). These fungi have been subcultured on PDA + chloramphenicol for about two years without any issue, but do not grow anymore when subcultured. Those fungi belong to the genera Alternaria, Pestalotiopsis, and Trichoderma. We are incubating them under the same conditions as before, at 20-21°C, and with the same culture medium, so I don´t really know what went wrong. Any recommendations?
Add host plant leaves and stem extract in media. Try and see results.
Add host plant leaves and stem extract in media. Try and see results.
Hi,
You can use leaves extract by water after freeze drying of extract
It could be that in those 2 years the pure cultures of the fungi I suppose were not properly stored; that is they were not fridge-stored at about 4-6 degrees Centigrade. In that case, the isolates may have run out of nutrients from the medium (PDA). You need to try resusitate them by subculturing them on potato avocado leaf or stem broth (PAB). If they do not grow then it may be that are dead or seriously moribund. Another thing is what dose or concentration of chloramohenicol did you add to the PDA? If the dose is high, being antibiotic, chances are it could antagonize the fungal isolate over time. Or may result in nutrient adsorption and unavailability to the isolates. Hence, both the hyphae and spores of the organisms will shrivel and die especially in the face of high antibiotic concentration.
It could be that in those 2 years the pure cultures of those fungi I suppose were not properly stored; that is they were not fridge-stored at about 4-6 degrees Centigrade. In that case, the isolates may have run out of nutrients from the medium (PDA). You need to try to resusitate them by subculturing them on potato avocado leaf or stem broth (PAB). If they do not grow then it may be that are dead or seriously moribund.
Another thing is what dose or concentration of chloramohenicol did you add to the PDA? If the dose is high, being antibiotic, chances are it could antagonize the fungal isolate over time. Or may result in nutrient adsorption and unavailability to the isolates. Hence, both the hyphae and spores of the organisms will shrivel and die in the face of high antibiotic concentration.
Dear Frédérique Reverchon .
You can use an extract of plant parts such as leaves , stems and roots by adding them with the media and then see the fungus growth.
Thanks a lot to you all for your insightful answers. We wil try to add plant extracts to the medium and see how it goes. Thanks again!
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