| Honey Treated Spore Tek
Sorry if this has been discussed before, but many years ago a FOAF discovered an interesting means of decontaminating heavily contaminated Ps. cubensis spores. My FOAF had received a dried cap (collected in southern Mexico, near the site of Wasson's original 'discovery'). Initial attempts at obtaining pure culture (on various agar media, including various antibiotic admixtures) were unsuccessful. During a search of professional journals however, my FOAF came across an article (originally published in Czech, as I recall) that described a process for decontaminating various wild collected mushroom spores by suspending them in honey. In the article, the decontaminating effect was attributed to the natural antibiotic quality of honey (which accounts for its resistance to developing mold or bacterial infections). After a little playing around with this technique, my FOAF was successful finally in isolating several (dikaryotic) cultures on regular MEA from these spores, yielding strains that fruited, vigorously, on rye cake media.
Basic Tek:
1) Mix a small quantity of spores (size of a pencil dot or two is enough) in a small quantity of fresh honey (my FOAF used fireweed honey, but other varieties seemed to work as well) in a small, sterile culture tube (15x125mm works well). Using a small rounded-head rod (a glass rod or even the handle of a streaking loop works well) crush and mix the spore material into the viscous honey. Make sure the spore clumps are completely broken up and well distributed throughout the honey.
2) Leave for 30 - 60 minutes (actually 15 minutes seemed to work as well, but the extra time provides extra insurance).
3) Place a small quanty (1-3 loopfuls) in a small quanttity (~5ml) of sterile water in another culture tube. Mix well to dissolve the honey/spore suspension. Allow to rest for 15 - 30 minutes, agitating periodically (swirl the tube) to separate and hydrate the spores.
4) Streak (using standard micro cross-streaking technique) on MEA or other nutrient agar plates.
* If the colonies from step 3 are still too dense (not isolated), perform serial dilutions (tranfer a few loops from step 3 into another 5 ml tube of sterile water) and re-streak using the diluted spore suspension. Repeat dilutions as required to obtain isolated colonies.
It's been a number of years since my FOAF tried this tek - and only on one Ps. cubensis specimen - so I can't vouch for its general efficacy (or for my accurate recollection of all the details), so YMMV..
-TheDart
| 
03-16-07, 09:37
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