Review the conditions you are using for the maintenance of your Monilinia fungus might be warranted.

When nutrient rich media is used and the photoperiod and temperatures are not right the sporulation can be affected.

I find I get better results which I grow on more natural media and conditions. If you had used a common media like potato dextrose agar I would try nectarine dextrose agar with a low dextrose content.

Make sure your conditions have a 12 hour light cycle which helps many fungi to be stimulated to sporulate.. Try storing your isolates under conditions such as sterile distilled water and in soil and test how they regenerate.

I looked at some Dutch research and they find robust sporulation at 10C on pear fruit but reduced sporulation at higher temperatures.

Mohammad,

First, confirm by PCR test or by DNA sequencing that you still have Monilina strain. Contamination often occures with such fungi. Do everething suggested by Paul, and try low doses of UV light at low temperatures.

Alex

Hello, better media for sporulation of

Monilinia spp. is apple juice agar.

Use 12 hour light and 12 hour dark for simulation of sporulation, near UV light promotes sporulation of this fungus.

Dear Dr. Paul Reed Hepperly ,

Thanks for detail response.

But how i can prepare a nectarine dextrose agar media? Is it enough to replace nectarine with potatoes in PDA media preparation?

Could you please send me the any file or link of this Dutch research?

dear Alex Ignatov

Thanks for the response and advice

Our fungus is already precisely identified and I'm sure it's the monilia fructigena.

dear Karen A. Darbinyan

Thanks for response.

But in the previous answers Dr. Paul Reed Hepperly suggested the nectarine dextrose agar media and I do not know which one to choose.

Is there a scientific source for describing the specific media of this fungus?

Van Leeuwen, G., Hobb, I, and M. Jeger. 2002. Factors affecting mummification. Mol. Plant Path. Open Access December 20, 2002. Prepare you nectarine dextrose agar using diced nectarines that are cooked and strained. Use half the dextrose of PDA and agar to your solification need. Hope the information is useful. In the Dutch study the low temperature favoring sporulation was apparent this is outside the normal media growth temperatures. UV light can be very important as Alex noted.

Dear Dr. Paul Reed Hepperly ,

Thank you for response quickly and providing such wonderful explanations

Best Regards

You can inoculate autoclaved pieces of potato tubers (without skin). Take shapes like for pommes frites (French fries). The sporulation on them is usually abundant.

You may try inoculation on the original host and isolate again. Sometimes fungi lost sporulation when you subculture them for many times on agar media.

For good sporulation, media, pH, diffusing light, host healthy extract play vital role. Scarcity of nutrients in media also stimulates sporulation.

You could try with PDA amended with tomato and photoperiod. But if you are sure that the isolate is M. fructigena better inoculate apple/peach...fruit to recover fresh conidia