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 Joshua
Holoman
Registered: 10/27/98
Posts: 5,398
Loc: The Matrix
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 First time w/ agar. 
#483775 - 12/08/01 03:08 AM (22 years, 9 months ago) |
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I have for the first time attempted to start mycelium from agar-agar. I thought of a few questions while innoculating.
I used agar-agar. Does agar-agar have nutrients sufficient for mycelial growth or is it just a gelling agent?
When innoculating I innoculated 20 sites w/i the dish. I tried to apply spores at uniform locations. Is this the proper method? If not, what is the best method?
I will now store my dishes in a dark warm location.
Any additional advise for a first-timer at useing agar would be appreciated.
Thank you,
Joshua
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 MAIA
World-BridgerKartikeya (DftS)
Registered: 04/27/01
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 Re: First time w/ agar. [Re: Joshua]
#483807 - 12/08/01 05:08 AM (22 years, 9 months ago) |
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"I used agar-agar. Does agar-agar have nutrients sufficient for mycelial growth or is it just a gelling agent?"
It's just a gelling agent to creat a solid medium, it's has a poor nutricional value, that's why you have to add Maltose or Dextrose, that is if you wish to follow a tested preparation (MEA or DEA).
Peace,
MAIA
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Spiritual being, living a human experience ... The Shroomery Mandala
 
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40oz
Registered: 01/18/01
Posts: 30,119
Loc: Sandy Eggo. Ca.
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Re: First time w/ agar. [Re: Joshua]
#484252 - 12/08/01 04:45 PM (22 years, 9 months ago) |
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...me thinks you fuct up...
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Joshua
Holoman
Registered: 10/27/98
Posts: 5,398
Loc: The Matrix
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Re: First time w/ agar. [Re: Joshua]
#484299 - 12/08/01 05:36 PM (22 years, 9 months ago) |
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Alright I messed up.
How about the rest of my post, innoculation procedure and general advice? (advice other than don't mess up again;)
Joshua
--------------------
The Shroomery Bookstore
Great books for inquiring minds!
"Life After Death is Saprophytic!"
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felixhigh
Scientist
Registered: 06/24/01
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Loc: Ly
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Re: First time w/ agar. [Re: Joshua]
#484502 - 12/08/01 09:31 PM (22 years, 9 months ago) |
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make several media dishes and leave them in the fridge so that you'll always have a 'virgin' dish as you need. as soon as new growth is seen you should transplant to other dish(es). against contaminants you can use gentamicyn and/or H2O2...
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40oz
Registered: 01/18/01
Posts: 30,119
Loc: Sandy Eggo. Ca.
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Re: First time w/ agar. [Re: Joshua]
#484666 - 12/09/01 12:10 AM (22 years, 9 months ago) |
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....its only a mistake if it happens twice...;)
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mycophyl
journeyman
Registered: 09/09/01
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Re: First time w/ agar. [Re: Joshua]
#484677 - 12/09/01 12:26 AM (22 years, 9 months ago) |
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here are a cpl of links to agar info that will help you get started http://www.shroomery.org/findorgrowthem/culturemediums.html http://www.shroomery.org/findorgrowthem/fgagar.htm good luck
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Joshua
Holoman
Registered: 10/27/98
Posts: 5,398
Loc: The Matrix
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 Re: First time w/ agar. [Re: Joshua]
#484786 - 12/09/01 03:20 AM (22 years, 9 months ago) |
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Any ideas on how to save my work?
I was thinking maybe pouring just above melting temp MEA over my plates.
Joshua
--------------------
The Shroomery Bookstore
Great books for inquiring minds!
"Life After Death is Saprophytic!"
Edited by Joshua (12/09/01 03:24 AM)
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altarego
member
Registered: 10/25/01
Posts: 130
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Re: First time w/ agar. [Re: Joshua]
#484831 - 12/09/01 06:30 AM (22 years, 9 months ago) |
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Joshua -
Just because agar lacks nutrients does not mean the spores won't germinate (H2O is all they need.) In fact, a nutrient free media is good for strain storage, since it slows growth without stopping it.
I would let them germinate and then transfer to MEA or PDA plates soon afterward.
- AE
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Trail_Blazer
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Re: First time w/ agar. [Re: altarego]
#484833 - 12/09/01 06:37 AM (22 years, 9 months ago) |
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They must need then just water, otherwise you would have hundreds of spore syringes out there right now germinating and creating tiny mycelium netoworks. I believe spores need both water and SOME form of nutrient to germinate. Be that nutrient, grain, birdseed, or even honey (think it uses the dextrose or glucose, i'm no chemistry/biology major here).
Correct me if i'm wrong, but i'm 99.9% sure spores won't germinate with water alone.
Peace
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altarego
member
Registered: 10/25/01
Posts: 130
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Re: First time w/ agar. [Re: Trail_Blazer]
#485147 - 12/09/01 02:51 PM (22 years, 9 months ago) |
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From my mycology textbook, what I can gather is that "germination" is a loose term, as even spores are in a very low (but not zero) rate of metabolism.
Here's a quote (from Deacon, _Modern Mycology_, 1997, p. 159): " . . . all spores germinate in essentially the same way. The cell becomes hydrated, there is a marked increase in respiratory activity, followed by a progressive increase in the rates of protein and nucleaic acid sythesis. An outgrowth (the germ-tube) is then formed, and it [. . . ] develops into a hypha . . ."
Now I take this to mean that up to the germ tube stage, no external nutrients are needed, meaning also that all those spores floating around in your syringes HAVE germinated, but are still "dormant" needing nutrients to grow any further.
The question is whether agar alone might have sufficient nutrients for this, and that I do not do. Agar is a protein mixture, so the fungus might be able to utilize it, although probably rather poorly.
As I said to Joshua in a PM, he might want to transfer the innoculation points on the agar plates to MEA or the equivalent.
Or start over.
Best,
AE
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altarego
member
Registered: 10/25/01
Posts: 130
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Re: First time w/ agar. [Re: Trail_Blazer]
#485149 - 12/09/01 02:54 PM (22 years, 9 months ago) |
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Joshua
Holoman
Registered: 10/27/98
Posts: 5,398
Loc: The Matrix
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Re: First time w/ agar. [Re: altarego]
#485360 - 12/09/01 07:39 PM (22 years, 9 months ago) |
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Thanks AE.
I don't think that I will transfer th innoculation points as I have 20 per dish. It would take too much time and I am not in short supply of spores. I will experiment with my mess up and post what my results are. If anyone knows the melting temp of agar I would appreciate it. Also I need to know the max temp spores can be subject too with out a marked decrease in viability. I will look for this info on my own, however it would save time if someone already knows the answer.
Joshua
--------------------
The Shroomery Bookstore
Great books for inquiring minds!
"Life After Death is Saprophytic!"
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