To search for new species that can be cultivated we make some general recom-mendations that the senior author P. Callac has realized from decades of research in mushroom growing:
1) If several sporocarps are found from the same site make isolates from at least two.
2) If you have a good specimen make at least two isolates, one from the cap and one from the stipe; all isolates must have different identification code numbers even if they come from the same piece of cap of the same sporocarp.
3) Any isolate growing very slowly or developing sectors with different aspects or growth rates can be discarded immediately.
4) Although the optimal conditions for each species is unknown, where possible strain isolation must be made on a culture medium similar the one that will be used to maintain the strain over many years.
5) Avoid taking too much precaution against potential contamination; for example in most cases it is not necessary to use antibiotics in the culture medium to isolate a strain and it is not necessary to use very thin slices of mushroom tissue. Thus the strain may be less fastidious in it sterile requirements when producing fruiting bodies.
6) Obtaining a good isolate is more important than obtaining a spore print, since the isolate will allow you to get a spore print from a future fruit body, while the spore print will never allow you to get the parental strain; however a quickly made spore print from the wild specimen (= the parent) can save time when performing hybridization.
7) For any isolate that successfully passes the fruiting test and which is maintained on non-synthetic medium (such as compost extract medium for Agaricus), make in parallel a mycelium growth rate test on a synthetic or semi synthetic reproducible medium (for example 14 days-test on malt agar medium). The growth rate is characteristic of the strain that will be further improved.
8) Each retained strain should be conserved through at least two independent clonal lineages.