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Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
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Needs investigation, Cubensis: oxidative enzymes and defenses. 1
#26915915 - 09/03/20 04:14 PM (3 years, 4 months ago) |
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Here is something that needs legal investigation, and someone with access to a LAB. I managed to get C. Burma to decay and grow on plant phenols, absent of common sugars. The result for me, after a few transfers on T-Gel, was the C. Burma was capable of decaying and converting plant phenols as it's carbon source.
Normally Cubensis, and other fungi, require a glucose source, from sugars, starch, cellulose and a few other materials.
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T-Gel, is just simply black tea and water, steeped as normal, and the bag is removed (99% is removed).
https://frida.fooddata.dk/food/115?lang=en || https://nutritiondata.self.com/facts/beverages/3967/2
As you can see from the links, tea, ready-to-drink contains no notable sugar, starch or cellulose.
Phenolic content in tea: https://en.wikipedia.org/wiki/Phenolic_content_in_tea
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First I germinated C. Burma on a wood peg with trace sucrose, placed into WL-Tek (paper + soluble fertilizer).

The first transfer to T-Gel, grew, but weak-ish, and tried to pin after a few weeks.

The second transfer was more vigorous, and produced visible white-rot (bleaching).

When left for a long period, and then exposed to oxygen, it lightly bruised.

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http://www.davidmoore.org.uk/Assets/Mostly_Mycology/Lucy_Goodeve-Docker_bioremediation_website/whiterotfungi.htm
"The lignin-degradative system of Phanerochaete chrysosporium appears after cessation of primary growth (that is, it is an aspect of the secondary metabolism of the organism) and can be induced by nitrogen starvation."
Primary and secondary carbon sources (reversed after plant production): https://imgur.com/a/8jngxe8
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I would like clarification of the enzymes C. Burma secreted on T-Gel. And if increasing oxidative decay, increases antioxidant production. Psilocybin is apparently a primitive antioxidant, therefor increasing oxidative stress should increase production.
https://en.wikipedia.org/wiki/Psilocybin (see chemistry and biosynthesis) https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3488923/
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In addition, the sample produced new cells, and potentially sugars from phenolic decay (it appears).
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Munchauzen


Registered: 06/22/11
Posts: 14,342
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Re: Needs investigation, Cubensis: oxidative enzymes and defenses. [Re: Ferather]
#26915945 - 09/03/20 04:37 PM (3 years, 4 months ago) |
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Why call it t-gel if its just tea?
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Roger Clemency
Smile


Registered: 03/23/20
Posts: 2,005
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Re: Needs investigation, Cubensis: oxidative enzymes and defenses. [Re: Munchauzen]
#26915964 - 09/03/20 04:50 PM (3 years, 4 months ago) |
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I feel like I’ve seen this before...and someone said ‘Is that different from starved agar?’ Idk what starved agar is or what’s up here but I couldn’t find this posted elsewhere. Am I still real?
-------------------- Sour grapes, sweet revenge Heaven starts right where hell ends
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A.k.a
Stranger



Registered: 10/27/19
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Loc: Gaming the system
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Re: Needs investigation, Cubensis: oxidative enzymes and defenses. [Re: Roger Clemency]
#26916039 - 09/03/20 05:31 PM (3 years, 4 months ago) |
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lol yeah that was me.
It’s just agar no nutes.
I think this needs to go to advanced myco to have any hope of discussion for you op.
Edited by A.k.a (09/03/20 05:32 PM)
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Munchauzen


Registered: 06/22/11
Posts: 14,342
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Re: Needs investigation, Cubensis: oxidative enzymes and defenses. [Re: A.k.a]
#26916191 - 09/03/20 06:41 PM (3 years, 4 months ago) |
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oh its tea and agar... lol. ok I was confused, maybe a bit baked 
I tried to google it and all I got was shampoo
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Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
Last seen: 1 year, 2 months
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Re: Needs investigation, Cubensis: oxidative enzymes and defenses. [Re: Munchauzen]
#26917246 - 09/04/20 11:03 AM (3 years, 4 months ago) |
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@ Munchauzen, T-Gel, yes as you worked out, just tea and agar (gelling agent). You can call it Tea agar.
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@ A.k.a, its not strictly starved, there more than enough nutrients including nitrogen. https://frida.fooddata.dk/food/537?lang=en
As you can see from the provided link, dry tea 100g, is richer than most grain (3.1% nitrogen, almost 20% proteins). It has roughly as much carbohydrates, but its nearly all cellulose (fiber) not starch or sugars.
In fact tea waste can be used to enrich wood-lover grows, and get more yield.
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You can compare tea, ready to drink (see original post) and 3g ME to 100g water.
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Here is C.Burma on a 'used' tea bag, germinated on a wooden toothpick peg.

No starch and trace sugars, got bleaching (white-rot).
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Quote:
A.k.a said: I think this needs to go to advanced myco to have any hope of discussion for you op.
Possibly, but some of the data provided will be enough for some people, and more people visit this section.
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Here is a sample fruit body of Cubensis, grown on WL-Tek, using brown rice spawn, with max FAE (maximum atmospheric oxygen). Same as what is normally done, grain spawn + coir, but instead of coir, WL-Tek paper pellets, no lid when fruiting.

The last two images are a single fruit, with the stem and a section cut.
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Recipes: WL-Tek (no tea), T-Gel
Edited by Ferather (09/04/20 01:18 PM)
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ttching8475
Spawn

Registered: 03/01/18
Posts: 739
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Re: Needs investigation, Cubensis: oxidative enzymes and defenses. [Re: Ferather]
#26919376 - 09/05/20 01:40 PM (3 years, 4 months ago) |
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Here is p cubensis RW and a competitor mold on a black tea and potato agar. Recipe was 1 black tea bag steeped in 200 ml water. 125 ml of the resulting tea was used with 5 ml potato flakes and 2.5 g agar powder. No added sugars and half the amount of potato from pasty’s pda recipe.

The transferred culture was on standard pda. The additional acidity of the t-agar seems to have ‘melted’ the pda a bit and the expanding myc is nearly translucent, but clearly very organized and expressive of the culture. Here it is on pda a few transfers back:
-------------------- LAGM 2.022
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Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
Last seen: 1 year, 2 months
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Re: Needs investigation, Cubensis: oxidative enzymes and defenses. [Re: ttching8475]
#26919489 - 09/05/20 02:35 PM (3 years, 4 months ago) |
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The mold says you need to be more aseptic, its come from air, or as you added the wedge. Mold will germinate both on the potato dextrose, and tea phenols (you can get mold growing in-on cups of old tea) and the tea bag.
I can see the phenotype has changed, the original is highly tomentose, I don't think your RW likes potato dextrose, or you have a competitor. Switch to malt extract.
Mycolorado's (TC) alternative T-Gel recipe: 100g > Water | 2g > Agar | 2g > Black Tea | 1g > Malt Extract.
Transfer until growth looks like mine or Mycolorado's.
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ttching8475
Spawn

Registered: 03/01/18
Posts: 739
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Re: Needs investigation, Cubensis: oxidative enzymes and defenses. [Re: ttching8475]
#26919620 - 09/05/20 03:38 PM (3 years, 4 months ago) |
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This is a second sample, which is clean so far but has more condensation in the no pour. My current work area is subject to temperature swings and airborne competitors, a challenging agar environment, but a good opportunity to fail and dial in the best practices.
In fairness to the comparison, this is an old culture and the pda picture is somewhat earlier in it’s life. I successfully fruited that generation. With that said, organization has been very consistent across transfers and that’s the best picture I have of it. The culture is VERY tomentose on pda and grain. So was the parent culture, an F9.
-------------------- LAGM 2.022
Edited by ttching8475 (09/05/20 03:41 PM)
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Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
Last seen: 1 year, 2 months
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Re: Needs investigation, Cubensis: oxidative enzymes and defenses. [Re: ttching8475]
#26920960 - 09/06/20 10:55 AM (3 years, 4 months ago) |
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Quote:
ttching8475 said: ...The culture is VERY tomentose on pda and grain. So was the parent culture, an F9.

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So some thoughts, When I tried germinating C.Burma on rye, I got tomentose growth immediately. But when I germinated as above (original post), I got healthy growth. There's essentially 3 main phenotypes, tomentose (woolly, matted), normal (populated, linear), and rhizomorphic (aggregated, clumped).
The growth from the peg in WL-Tek, is normal and highly populated (substrate disappearing). The growth on T-Gel, first transfer, is normal but some malnutrition (not adapted). The growth on T-Gel, second transfer, is normal and highly populated (no gaps).
Transfer of a wedge to brown rice, started as normal but some malnutrition (first time on starch), than after 2-3 days, normal and highly populated.

Note: I did not use malt extract in my T-Gel tests, only tea.
Edited by Ferather (09/06/20 11:32 AM)
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