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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: Anglerfish]
    #27557350 -

:whathesaid:


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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: atomicshark] * 2
    #27557378 -

rhizoRider said:
Don’t op pics lol like subsectoids? Or a variant close to them. ?



The spores are too narrow for those.

atomicshark said:
Was anyone able to to genetic analysis?



I would be happy to sequence them.


Quote:
Nevertheless, do you have any other hints for identification which i can conduct?



I suggest using Piximetre to measure 30 spores, 10 cheilocystidia and 10 pleurocystidia, if present.  For the cystidia do a crush mount.  Measure spores at 1000x and cystidia at 400x.

When looking at the gill cystidia remove a piece of gill with a forceps and set it down on the glass side, paying attention to which side is the edge.    Use a brand new razor blade to cut just the edge off, giving you a piece about .1 mm x .5 mm, and mount that and do a crush mount.  Cut the remaining piece of the gill in half and throw away the part that was closest to the gill edge, as this could have a mix of cheilocystidia and pleurocystidia.    Mount the part of the gill that was furthest from the gill edge so you only have pleurocystidia, basidia and lamellar trama elements in that view.  For the crush mounts press down firmly enough to break the cells apart but not firmly enough to break the spores.

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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: Alan Rockefeller]
    #27557393 -

Alan Rockefeller said:
When looking at the gill cystidia remove a piece of gill with a forceps and set it down on the glass side, paying attention to which side is the edge.    Use a brand new razor blade to cut just the edge off, giving you a piece about .1 mm x .5 mm, and mount that and do a crush mount.  Cut the remaining piece of the gill in half and throw away the part that was closest to the gill edge, as this could have a mix of cheilocystidia and pleurocystidia. Mount the part of the gill that was furthest from the gill edge so you only have pleurocystidia, basidia and lamellar trama elements in that view.  For the crush mounts press down firmly enough to break the cells apart but not firmly enough to break the spores.


:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

And for caulocystidia and pileipellis ? How to create good perspective ?


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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: RenegadeMycologist]
    #27557518 -

I modified a python script which was able measuring the dimensions of spores automated.


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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: RenegadeMycologist]
    #27557524 -

That is 7 spores and dimensions are in pixels. If you think we're gonna calculate that one by one to get physical dimensions, you're wrong. Please update your script to show physical dimensions and measure at least 30 spores.
Meanwhile I will provide you with reference material.


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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: RenegadeMycologist]
    #27557526 -

According to the:

Borovička J. (2008): The wood-rotting bluing Psilocybe species in Central Europe – an identification key. – Czech Mycol. 60(2): 173–192.

"Spores in P. arcana are of the same size as in P. serbica, (9.8–)10.5-12.5(–13.5) × (6.2–)6.5–7.1(–7.4) μm [Q = 1.5–1.7–1.85]. Basidia 4-spored, with relatively short sterigmata (3.5–5 μm)."

Borovička also mentions other identifing microscopic features for arcana:

"Pleurocystidia absent. Cheilocystidia lageniform, mostly with a relatively short neck (up to 10 μm)."

Scent of arcana in the same paper is described as: "radish-like".
And habitat: "...mostly in broadleaved forests (Quercus, Fraxinus, Acer), but also under conifers (e.g., Pinus)."

Original (detailed) macro/micro description of pelliculosa can be seen here:
http://www.cybertruffle.org.uk/cyberliber/59350/0050/002/0280.htm


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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: RenegadeMycologist]
    #27557550 -

RenegadeMycologist said:
That is 7 spores and dimensions are in pixels. If you think we're gonna calculate that one by one to get physical dimensions, you're wrong. Please update your script to show physical dimensions and measure at least 30 spores.
Meanwhile I will provide you with reference material.


Yes, I'll count over 30 spores next time I'm in the lab. And the script is still a little buggy and needs improvement.

Edited by atomicshark (11/25/21 01:40 PM)

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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: atomicshark]
    #27557594 -

Here you could find a slightly improved version of script.
Feel free to implement additions.
https://pastebin.com/nGPa6cbL


Edited by atomicshark (11/25/21 02:27 PM)

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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: RenegadeMycologist]
    #27557663 -

RenegadeMycologist said:
And for caulocystidia and pileipellis ? How to create good perspective ?


Caulocystidia and pileipellis aren't commonly used to differentiate Psilocybe species, but will be useful in other genera or for making a full species description of Psilocybe.

For caulocystidia use a brand new razor blade to take an extremely thin scalp section about 2 mm below the gill attachment, mount in 5% KOH, press down firmly on the coverslip.

For pileipellis use a well dried cap and brand new razor blade and make lots (ten to twenty) of extremely thin cross sections, cut on a slight angle so one side is extremely thin and the other side is a bit thick.  Select a few of the thinnest ones, cut off and throw away the thick side leaving only the extremely thin part.

Most Psilocybe species have a gelatinized pileipellis (ixocutis) so don't expect to see anything dramatic like you would in some genera.

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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: Alan Rockefeller]
    #27557687 -

Alan Rockefeller said:
RenegadeMycologist said:
And for caulocystidia and pileipellis ? How to create good perspective ?


Caulocystidia and pileipellis aren't commonly used to differentiate Psilocybe species, but will be useful in other genera or for making a full species description of Psilocybe.

For caulocystidia use a brand new razor blade to take an extremely thin scalp section about 2 mm below the gill attachment, mount in 5% KOH, press down firmly on the coverslip.

For pileipellis use a well dried cap and brand new razor blade and make lots (ten to twenty) of extremely thin cross sections, cut on a slight angle so one side is extremely thin and the other side is a bit thick.  Select a few of the thinnest ones, cut off and throw away the thick side leaving only the extremely thin part.

Most Psilocybe species have a gelatinized pileipellis (ixocutis) so don't expect to see anything dramatic like you would in some genera.


:vibin:

Thanks


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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: RenegadeMycologist]
    #27558440 -

Here are the results of a Python script that measured 44 spores:

Spore Sizes:
a µm x b µm
7.001 11.977
6.863 11.538
6.495 11.443
6.705 11.145
6.707 11.664
6.847 12.748
6.57 11.941
6.707 11.226
6.904 12.943
6.302 11.23
6.727 12.704
6.415 10.828
6.325 11.313
6.846 13.04
6.419 11.211
6.809 12.998
6.155 10.304
5.999 7.596
6.621 10.699
6.597 11.992
6.231 10.997
6.31 10.342
6.342 9.112
6.884 11.878
6.693 11.715
6.495 11.649
7.254 11.584
6.804 10.309
6.707 10.206
6.726 11.333
6.691 12.495
6.618 13.023
6.316 12.126
6.835 12.352
6.756 11.252
6.416 11.624
5.803 7.563
6.496 12.678
6.473 11.919
6.148 6.311
6.804 11.443
5.905 10.932
7.013 12.739
6.651 12.16

### RESULTS ###
num spores:  44
min a:  5.803 µm
max a:  7.254 µm
min b:  6.311 µm
max b:  13.04 µm
avg a:  6.576931818181818 µm
avg b:  11.324590909090908 µm
std deviation a 0.30030305887456676 µm
std deviation b 1.419263322325767 µm
ratio avg_a / avg_b:  0.5807655102933681

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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: atomicshark]
    #27558450 -

Here is a revised run in which severely deformed spores were removed:

7.001 11.977
6.863 11.538
6.495 11.443
6.705 11.145
6.707 11.664
6.847 12.748
6.57 11.941
6.707 11.226
6.904 12.943
6.302 11.23
6.727 12.704
6.415 10.828
6.325 11.313
6.846 13.04
6.419 11.211
6.809 12.998
6.155 10.304
6.621 10.699
6.597 11.992
6.231 10.997
6.31 10.342
6.342 9.112
6.884 11.878
6.693 11.715
6.495 11.649
7.254 11.584
6.804 10.309
6.707 10.206
6.726 11.333
6.691 12.495
6.618 13.023
6.316 12.126
6.835 12.352
6.756 11.252
6.416 11.624
6.496 12.678
6.473 11.919
6.804 11.443
5.905 10.932
7.013 12.739
6.651 12.16

############################## RESULTS ##############################
num spores:  41
min a:  5.905 µm
max a:  7.254 µm
min b:  9.112 µm
max b:  13.04 µm
avg a:  6.620365853658538 µm
avg b:  11.629560975609756 µm
std a:  0.2600984575396693 µm
std b:  0.8783054929606793 µm
var a:  0.06765120761451517 µm
var b:  0.7714205389649018 µm
ratio avg_a / avg_b:  0.5692704881588552

Edited by atomicshark (11/26/21 04:02 PM)

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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: atomicshark]
    #27558506 -

Spore size tells nothing after all. I still think the distinction could be made on the basis of habitat in favor of pelliculosa.

Send it to Alan for sequencing, you might have first ever recorded find of this species for your country.


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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: RenegadeMycologist]
    #27559487 -

Today i tried to make some other pics of gills. Nevertheless, I've to improve microscopic sample preparing techniques.


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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: atomicshark]
    #27559582 -

atomicshark said:
Today i tried to make some other pics of gills. Nevertheless, I've to improve microscopic sample preparing techniques.




What part of the gill are we looking at here ?
You need to label your micrographs like in the first post, so we know what are we looking at.

Gill here obviously exibits abundant cystidia. You need to check for pleurocystidia now since spore size didn't yield any results. Hopefully there aren't any and this is settled. Consider aquaduct section if you have fresh specimens for best view of pleurocystidia.


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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: RenegadeMycologist]
    #27559599 -

I cut a slice out of the mushroom cap that looked like a comb. Then I covered it with water and compressed it in with another slide.

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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: atomicshark]
    #27559601 -

But are these last shots of the gill face or the gill edge?


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★★★★★

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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: Anglerfish]
    #27559606 -

Anglerfish said:
But are these last shots of the gill face or the gill edge?


Was was made from edges, but could also contain some faces through compressing.

Here is an another image which captures with a 10x objective a cap cross-section. I'm not sure from which position this was made, because the sample was to much deformed by compressing.
It was dyed with Safranin T.


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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: atomicshark]
    #27560288 -

There is another possibly interesting aspect: I was not able to get a spore print that was visible to the naked eye from around 50 specimens. I tried it with increased and reduced humidity.

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Re: Wood loving Psilocybe Mushroom - P. pelliculosa? [Re: atomicshark]
    #27560298 -

Well, the gills in your pictures appear to be quite pale, could indicate low spore production.
These also appear to be quite similar to P. semilanceata, which can be hard to print as well.
My experience with the latter species is that the larger and more open caps leave better prints.
The smaller ones with incurved margin not so much.


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