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Workman
1999 Spore War Veteran


Registered: 03/01/01
Posts: 3,593
Loc: Oregon, USA
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Mexicana Sclerotia Invitro
#474885 - 11/30/01 02:02 PM (22 years, 15 days ago) |
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As some of you know I have been working on an in-the-jar sclerotia forming strain of Psilocybe mexicana. It has taken awhile but so far the preliminary results are promising.
The strain fruits sparsely but it does fruit and produce some spores.

Below is a picture of two test jars at 3 months of age after inoculation. The sclerotia was still growing well at this time but I got tired of waiting.

Below is a closeup of one of the larger sclerotia pressing into the glass. This is a mature one with little to no recent growth evident. Actively growing sclerotia is white on the outer growing edge. This sclerotia may start growing again if left alone. Some actively growing spots in the jars can be seen in the 2nd image from the top.
 This is the left hand jar pictured above harvested. The waste seed is on the left, the crudely cleaned sclerotia is on the right. I would clean this a bit better before ingesting. There are still a fair amount of clinging grass seeds on them. You can see that about 1/2 the total jar volume is sclerotia. I didn't expect that much judging by what I could see from outside the jar.

I put my hand in the shot to show the relative size of one of the larger sclerotia. There were large amounts of pea sized sclerotia that were a bit tedious to harvest from the seed. The pile weighed in at 132 grams fresh weight from 1 cup of grass seed. Sclerotia is supposed to be about 30% dry matter so it should be nearly 40 grams dried. I am going to plant the larger sclerotia in a casing to generate more spores.
Edited by Workman (08/24/05 09:41 AM)
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Workman
1999 Spore War Veteran


Registered: 03/01/01
Posts: 3,593
Loc: Oregon, USA
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#474960 - 11/30/01 03:00 PM (22 years, 15 days ago) |
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I forgot to mention that the jars were incubated at room temperature (68-72F) in my kitchen on a top shelf with each jar completely covered in foil. The foil keeps the jars dark which is supposed to promote sclerotia formation. I peeked all the time so some light exposure doesn't seem to be harmful or inhibiting. I looked through "The Mushroom Cultivator" by Stamets and Chilton regarding sclerotia yeild from Psilocybe mexicana and compared it to my yeild. Note that this is a single jar test and may not be typical. The Mushroom Cultivator states: 50-70 grams per cup of seed at 2 months Spore Works test jar 132 grams per cup of seed at 3 months. I used 134 grams dry of grass seed (about 1 cup) and the cleaned and dried sclerotia from the test jars should dry to around 35 grams. That means over 25 percent of the dry seed mass was turned into dry sclerotia. I find that nearly unbelievable!
-------------------- Research funded by the patrons of The Spore Works Exotic Spore Supply My Instagram Reinvesting 25% of Sales Towards Basic Research and Species Identification 
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Azure
old hand
Registered: 12/31/98
Posts: 469
Loc: California, USA
Last seen: 21 years, 7 months
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#474984 - 11/30/01 03:23 PM (22 years, 15 days ago) |
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From what I've seen with mexicana sclerotia from your prints is that a good 30-40% of it is grass seed after it's cleaned thoroughly. With any further cleaning, you'd lose some of the goodies. None of pieces seem to be pure compressed mycelium, so that could cause an error in your weight measurements. I also understand that Stamets' sclerotia was probably a good 30-40% grass seed as well... Also, are you talking about dry weight or wet?
Edited by Azure (11/30/01 03:26 PM)
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Workman
1999 Spore War Veteran


Registered: 03/01/01
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Loc: Oregon, USA
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Re: Mexicana Sclerotia Invitro [Re: Azure]
#475001 - 11/30/01 03:34 PM (22 years, 15 days ago) |
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Of course these results are preliminary. I'll work on cleaning the sclerotia and reweighing, then dry and weigh again. They appear to be fairly solid. It will be interesting to see how this turns out. I should note that my other jar "appears" to have more sclerotia in it and I will be harvesting it soon. Also, this strain of P. mexicana is not generated or isolated from the B strain but is from a different collection. Currently, the Spore Works has only offered the B strain which fruits well and forms sclerotia in cased trays but tends to be inferior for invitro sclerotia formation.
-------------------- Research funded by the patrons of The Spore Works Exotic Spore Supply My Instagram Reinvesting 25% of Sales Towards Basic Research and Species Identification 
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LuciferX
ape

Registered: 11/18/98
Posts: 731
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#475136 - 11/30/01 05:43 PM (22 years, 15 days ago) |
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neat
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Lana
Head Banana


Registered: 10/27/99
Posts: 3,109
Loc: www.MycoSupply.com
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#475139 - 11/30/01 05:47 PM (22 years, 15 days ago) |
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WOW, good job workman:) Haven't seen you in awhile. But its nice to see that you're staying busy. Don't be a stranger! Me
-------------------- Myco Supply - Distributors of Mycological Products http://www.MycoSupply.com The Premiere Source for Mushroom Growing Supplies. Visit us online or call us toll free
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40oz


Registered: 01/18/01
Posts: 30,119
Loc: Sandy Eggo. Ca.
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#475161 - 11/30/01 06:08 PM (22 years, 15 days ago) |
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Rock On Workman! whats the dosage on those babies?
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World Spirit
PNW



Registered: 07/27/01
Posts: 9,817
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#475258 - 11/30/01 07:45 PM (22 years, 15 days ago) |
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Deleted by admin
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B.I.O.
old hand

Registered: 05/26/99
Posts: 458
Last seen: 9 months, 13 days
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Re: Mexicana Sclerotia Invitro [Re: World Spirit]
#475365 - 11/30/01 09:13 PM (22 years, 15 days ago) |
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worky...thaz great man....wow....your jars look like mine that i use from now on....i use glassmilkbottles with a screwelid (polypropylene) ...all is autoclaveable and you can use filterdisks too...my all american can take something like 18 bottles at 1 time...the second one 12 or so.... BTW...i have to get in contact with some zapotec growers...i have some problems....pins wont develop further...also in the jars small scleros develop...im gonna post that soon BiO.
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World Spirit
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Registered: 07/27/01
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Re: Mexicana Sclerotia Invitro [Re: World Spirit]
#475374 - 11/30/01 09:18 PM (22 years, 15 days ago) |
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Deleted by admin
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Workman
1999 Spore War Veteran


Registered: 03/01/01
Posts: 3,593
Loc: Oregon, USA
Last seen: 22 hours, 23 minutes
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Re: Mexicana Sclerotia Invitro [Re: World Spirit]
#475486 - 11/30/01 10:54 PM (22 years, 15 days ago) |
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The lids are the off the shelf plastic lids you can get where you buy canning jars (i.e. Walmart). The say something like "not for food processing" but they can take the heat of pressure cooking and work great. You can drill or melt a hole in them for a filter disk or use a wad of polyfil in the hole as a cheap filter.
There is no birdseed in the jars. The jars are 2:1 grass seed to water.
I did try adding some dung to seed jars with poor results. I may try again as the problem was probably more with my fruiting method than the dung addition.
Taste is subjective but I have nibbled on a bit of fresh tampanensis sclerotia to check for flavor. It is very similar to fresh walnuts in texture and taste with a minor bitterness and fungal overtones. I have heard that it tastes worse the more you eat. I am not sure what the dry material tastes like but it is very dense and stony and would require a lot of chewing.
Ok, a little update on the crop. I carefully cleaned off all of the clinging grass seed and reweighed the sclerotia. They look much nicer cleaned off.
New total wet weight = 119 grams Estimated dry weight at 30% recovery = 35.7 grams
I also sliced one open to check for hollowness and encased seed. No seed present with only minor voids. They are basically solid.

I will write a more complete method and description and post it at the Spore Works some time in the next week or so.
-------------------- Research funded by the patrons of The Spore Works Exotic Spore Supply My Instagram Reinvesting 25% of Sales Towards Basic Research and Species Identification 
Edited by Workman (08/24/05 09:48 AM)
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Anonymous
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#475515 - 11/30/01 11:25 PM (22 years, 15 days ago) |
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Excellente workman, been waiting to see results of the other specimens you had to test. A question though. This was a culture started from a gill fragment, not from spore? Do you think this is a "culture only strain"? I sure hope you get some spores from any fruits you can milk from the sclerotia!!!!!!!! Bitter is an understatement at 3 + grams dry per dose!!!! Keeping my fingers crossed for SPORES!!!!!!!!!!!!!
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Workman
1999 Spore War Veteran


Registered: 03/01/01
Posts: 3,593
Loc: Oregon, USA
Last seen: 22 hours, 23 minutes
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Re: Mexicana Sclerotia Invitro [Re: ]
#475534 - 11/30/01 11:49 PM (22 years, 15 days ago) |
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It is originally from a gill fragment. The specimens were dry and the tissue was unviable. But the gills are loaded with dormant spores clinging to the gill tissue. So essentially what was cultured is not a clone of the gill tissue but a multispore culture, just like what you would get from a print. I have accumulated some spores from this culture.. It takes forever to fruit, produces a great deal of sclerotia in the casing, and then fruits sparsely. I plan on planting some of the freshly harvested sclerotia to see if that increases fruit production. This A strain produces a more conic cap as compared to the more plane cap of strain B. The original dried specimen of strain A has a distinct nipple that isn't present on the cultivated specimens generated from it.
-------------------- Research funded by the patrons of The Spore Works Exotic Spore Supply My Instagram Reinvesting 25% of Sales Towards Basic Research and Species Identification 
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Anonymous
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#475536 - 11/30/01 11:55 PM (22 years, 15 days ago) |
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From spores!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! Can't wait. Thank you for the timely response, and Let us know when they are available for purchase. Thanks Sorry for the cross post question at the Forest Floor!!!
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40oz


Registered: 01/18/01
Posts: 30,119
Loc: Sandy Eggo. Ca.
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#475591 - 12/01/01 01:08 AM (22 years, 15 days ago) |
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Bitch'n work, Workman! cant wait for your write up!
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Anno
Experimenter



Registered: 06/17/99
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#476035 - 12/01/01 12:05 PM (22 years, 15 days ago) |
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Workman, that?s a fantastic biological efficiency you achieved here !! You said you used 1 cup (236ml) of rye grass seed, which weighs app 75 g. From this 119 g sclerotia were produced and assuming that they contain 30% dry matter , you have 35,7 g dry matter. The definition of "biological efficiency" says that "achieving a 10% conversion of dry substrate mass into dry mushrooms" is 100% biological efficiency. (This definition was developed by the white button mushroom industry, since they achieve around 100% biological efficiency). So you have 35.7g/75g/0.10*100 = 476% biological efficiency, which is really a great result !! According to Stamets, "up to 50% conversion of wet substrate mass into harvestable mushrooms is possible". In comparison to that P. tampanensis that Una uses for cultivation of sclerotia yields around 50g wet per 110g of dry substrate, meaning 0.30*50g/110g/0.10*100 = 136%, which is still very good but pales in comparison with this mexicana. I?m very interested in the results of the other jar.
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Workman
1999 Spore War Veteran


Registered: 03/01/01
Posts: 3,593
Loc: Oregon, USA
Last seen: 22 hours, 23 minutes
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Re: Mexicana Sclerotia Invitro [Re: Anno]
#476049 - 12/01/01 12:22 PM (22 years, 15 days ago) |
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Not quite, I used 134 grams of dried seed. It compresses a bit and I packed it pretty tight and I didn't use exactly a one cup measurement so it was probably a bit over 250ml in volume. So lets put that number into your calculations. 134 grams dry seed 119 grams cleaned sclerotia (wet) Assumed 35.7 grams dry matter. 35.7/134/0.10*100 = 266.4% biological efficiency
-------------------- Research funded by the patrons of The Spore Works Exotic Spore Supply My Instagram Reinvesting 25% of Sales Towards Basic Research and Species Identification 
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Workman
1999 Spore War Veteran


Registered: 03/01/01
Posts: 3,593
Loc: Oregon, USA
Last seen: 22 hours, 23 minutes
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#476234 - 12/01/01 04:22 PM (22 years, 14 days ago) |
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Final numbers:
134 grams dry grass seed 2:1 ratio seed to water
Jar #1 119 wet grams dried to 40.7 grams = 34.2% dry matter Jar #2 108 wet grams
I would say a dry ounce (28 grams) per jar is a conservative estimate after 3 months of incubation.
The latest images are posted at:
Mexicana Gallery
There is no dry weight for jar #2 because I planted the fresh sclerotia in casing soil in order to encourage fruiting.
-------------------- Research funded by the patrons of The Spore Works Exotic Spore Supply My Instagram Reinvesting 25% of Sales Towards Basic Research and Species Identification 
Edited by Workman (08/24/05 09:55 AM)
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World Spirit
PNW



Registered: 07/27/01
Posts: 9,817
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#476386 - 12/01/01 07:17 PM (22 years, 14 days ago) |
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Deleted by admin
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psilocyber
old hand

Registered: 06/08/99
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Re: Mexicana Sclerotia Invitro [Re: Workman]
#487517 - 12/11/01 02:45 PM (22 years, 4 days ago) |
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Post deleted by administrator.
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