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Satori1
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P. semilanceata on agar attempt
#27072945 - 12/05/20 05:17 AM (3 years, 2 months ago) |
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A few P. semilanceata specimens were found in a Nordic country in late September, were air dried for a few days, kept at room temperature for a few weeks, then kept in freezer for a few more weeks. Then some dried gill tissue was sprinkled on top of malt extract +antibiotic (Aminoglycoside) plates and kept around room temperature for 5 days. There was skepticism that this would work, given the less-than-ideal conditions, but the paper cited here gave some hope that it was worth a shot.
Interestingly, some gill tissue was soaked in water for ~6 hours and then plated as described in The Mushroom Cultivator, but those plates have almost no growth. The photos below are from simply sprinkling gill tissue that was dried/frozen and cut onto agar. Contamination was expected at this stage on the particular plate in the images below (about seven-ten small worms that were wiggling around in the gill tissue of this specimen after the first day of drying had to be removed).
Others here and here have noted that P. semilanceata mycelium is very cottony, lacking rhizomorphic characteristics. The close-up images below seem to be a combination of both cottony and rhizomorphic growth. But maybe it is another species else entirely?
In any case, three agar pieces (see plate annotation image for sections that were removed) were cut and transferred to new plates of the same type today.
Any opinions about whether the cottony sections below could be P. semilanceata mycelium? Especially interested in hearing from those who have cultivated this species on agar before.
Guess there is no easy way to give a positive id. Was thinking of doing poor man's PCR, since P. semilanceata-specific primers have been published (article is in Polish, but one doesn't need Polish language skills to read the sequences )
*All images from the same plate. Brown specs are gill fragments*



This one looks the most promising, since this cottony/ropey growth is emanating from one of the gill fragments:


Very confused as to how the plates that received pre-soaked gill tissue display no growth, and the one that had gill tissue simply sprinkled on top is displaying various kinds of growth (both came from the same P. semilanceata specimen) Most interested, though, in identifying the white growth above.
**Edited for clarity
Edited by Satori1 (12/05/20 05:30 AM)
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wxorx
elsewhere


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Re: P. semilanceata on agar attempt [Re: Satori1]
#27073081 - 12/05/20 08:01 AM (3 years, 2 months ago) |
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I attempted agar cloning semilanceata too, and when something whitish appeared on agar I thought it is contaminated with some kind of white mold, but at the end it came out to be mycelium, here it how it looked on agar:

and grain:
-------------------- void **
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smalltalk_canceled
Babnik


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Re: P. semilanceata on agar attempt [Re: wxorx]
#27073130 - 12/05/20 08:38 AM (3 years, 2 months ago) |
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Quote:
smalltalk_canceled said: (original transfer pics, look at all this crazy mold shit that was in a 1/6 of a semilanceata spore print)

been working for a while now with isolating and cleaning up a wild sample of psilocybe semilanceata

-------------------- Willpower is the one true virtue
  
Edited by smalltalk_canceled (12/05/20 08:39 AM)
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Satori1
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Re: P. semilanceata on agar attempt [Re: wxorx]
#27076868 - 12/07/20 10:16 AM (3 years, 2 months ago) |
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Thanks for the pictures. Just out of curiosity, how did you determine it was mycelium? Because mushrooms grew out of it?
Also, what kind of grain is that?
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smalltalk_canceled
Babnik


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Re: P. semilanceata on agar attempt [Re: Satori1]
#27077055 - 12/07/20 12:13 PM (3 years, 2 months ago) |
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Corn or coriander.
I ended up assuming based on grow times and probably will never know for sure before I get a fruit. Which is considered impossible for new growers.
But I still have myc colonizing, it's just Hella slow
Hehe that's why I'm here, to see you wrestle the beast
-------------------- Willpower is the one true virtue
  
Edited by smalltalk_canceled (12/07/20 12:22 PM)
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Satori1
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After three agar transfers, two different isolates have been obtained from the original MS germination. They are called "A" and "B" and their identities are unknown. All incubation has been at about 18-22C. The following pictures are after three agar transfers with antibiotic-containing LME plates and photographed plates have been incubated for 8-9 days:
Characteristics:
Culture A: Very cottony/aerial/thickish white growth. After each transfer, it consistently grows for a few days from the transferred wedge then stalls after colonizing the middle 25-30% of the plate. It doesn't really expand much radially after that - but does seem to get thicker where it is already growing. Doesn't have any particular smell.



Culture B: Growth is very low on agar surface, and very very fast. Within 48 hours of transferring an agar wedge, 75% of the plate is colonized. The pungent odor of this culture is sweet and heavy, reminding me quite a bit of the smell of rotting fruit colonized by mold. As seen from the photos, this culture seems to establish an area of thicker growth immediately surrounding the transferred wedge, then displays thinner growth on the rest of the agar surface.



As a reminder, it's not clear what is being cultured here. The hope is that it is P. semilanceata.
The speed of growth and smell of culture B points to it being some kind of white mold. Any opinions?
Culture A appears to be more promising, but why doesn't it have the distinctive mycelial smell of, for example P. cubensis or P. ostreatus? Could it be that more mycelia is needed to produce that smell, as would be the case on grain? Haven't cultured either of those strains on agar before, so not sure what to expect in the way of odors ...
Any opinions about what is being cultured above, based on the pictures and descriptions would be greatly appreciated. Especially from those who've cultivated P. semilanceata on agar before What does it smell like at the stage where it's growing on agar?
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