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Joshua
Holoman


Registered: 10/27/98
Posts: 5,398
Loc: The Matrix
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Agar...what next? *pics*
#541960 - 02/06/02 12:19 AM (23 years, 18 days ago) |
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To start off with: for some of you this is a general cultivating question. However, I feel that agar culturing is an advance technique, that is why I am posting in this thread.
I have recently innoculated several agar plates w/ a multispore innoculation. I have fluffy mycelium as shown in the pics.
I am inquiring on my next course of action. I see no rhizomorphic mycelium on my plates. Should I transfer wedges to grain? or should I continue with a possible isolation? This is my first successful attempt at agar culturing and I am in need of advice.
In the future what would be my best course of action for culturing strong mycelium.
I could fruit from BRF and clone from there.
I could repeat my current process.
What other methods work well and will not waste my time?
Joshua
-------------------- The Shroomery Bookstore
Great books for inquiring minds!
"Life After Death is Saprophytic!"
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Anno
Experimenter



Registered: 06/17/99
Posts: 24,168
Loc: my room
Last seen: 9 days, 16 hours
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Re: Agar...what next? *pics* [Re: Joshua]
#542006 - 02/06/02 12:58 AM (23 years, 18 days ago) |
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Transfer a few small wedges from each dish to new plates. Then the growth should sector.
I allways go the "frut them once from multispore then clone a good looking fruit" path.
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altarego
member
Registered: 10/25/01
Posts: 130
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Re: Agar...what next? *pics* [Re: Joshua]
#542302 - 02/06/02 09:10 AM (23 years, 18 days ago) |
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Early plates are often too dense with strains to see secotring. Transfer very small wedges and the differences should become obvious.
I agree with Anno that cloning makes it much easier to get a pure strain, though it is fun to see strains emerge from the chaos of a plate.
See my new post for a followup question on this one.
- AE
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Suntzu
Geek


Registered: 10/14/99
Posts: 1,396
Last seen: 1 month, 20 days
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Re: Agar...what next? *pics* [Re: Joshua]
#542332 - 02/06/02 10:11 AM (23 years, 18 days ago) |
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Hey Joshua, welcome to 'advanced' I've read your other stuff and I like your spirit.
As usual, Anno is right.
Here's how I think about it. . .in that mottled mat of growth you have many many individual genetic combinations. Some non-mated/un-mate-able growth, some mated hyphae that lack clamp connections [but still might look like decent growth]. Those boundary zones are a good indication of multiple crossings.
Visualize hundreds of these potentials on a single multispore agar inoculation. If you put a chunk to grain or a chunk to agar, either way you've begun the process of weeding out the slower growing ones and if grain, the ones not capable of competing on the 'new' food source. . .whittling down the number to the ?couple/few? combos that are represented in the flush. With this species though, you'll usually get just fine results--no reason why you can't subculture to grain and agar! The whole 'rhizomorphic' thing is based on the observation that such growth is usually more aggressive and pins well; and if pure, will tend to pin more uniformly if cased/initiated right.
I've had senesced mycelium that was still rhizomorphic, though. Also, might be the generation, but the ecuador print I've used always gives fuzzy mycelium. . .even after repeated transfers, though it fruits admirably. Along the same lines, clones from fruits may not be as rhizomorphic as you'd think. . .at least on first transfer.
The way to do it, IMO, is to keep track of your cultures. When you get a nice flush, clone one or two. The more flushes you see, the more you'll be able to choose from. If possible, make slants or water stocks [easy] for LT storage; this gives you a library to go back to after observing a given flush. Just remember that unless it's pure, the mycelium growth will go through the selection process again upon the next expansion. Which could be a good thing, depending.
It's nice to have multispore growth in LT storage, though; you can basically go years without germinating spores.
One of the nicest strains I ever had was taken off a PF cake. I'm pretty sure it was pure; Ever notice how sometimes on a cake you'll get patches of very rhizo growth? Pair of tweezers, a lighter. . .
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Anno
Experimenter



Registered: 06/17/99
Posts: 24,168
Loc: my room
Last seen: 9 days, 16 hours
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Re: Agar...what next? *pics* [Re: Suntzu]
#542489 - 02/06/02 12:32 PM (23 years, 18 days ago) |
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>Pair of tweezers, a lighter. . .
Hehe, that?s exactly how I selected the culture for these Mazatapec.
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JuSMe13
newbie
Registered: 08/29/01
Posts: 25
Loc: california
Last seen: 22 years, 11 months
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Re: Agar...what next? *pics* [Re: Anno]
#542582 - 02/06/02 02:01 PM (23 years, 18 days ago) |
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Wow, I would love a print of those Babies!
very impressive
-------------------- Yo Quiero FSR !
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Seuss
Error: divide byzero


Registered: 04/27/01
Posts: 23,480
Loc: Caribbean
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Re: Agar...what next? *pics* [Re: Suntzu]
#542659 - 02/06/02 03:10 PM (23 years, 18 days ago) |
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Nice info, thanks! What are the benefits of slants vs petri dishes for long term storage?
-------------------- Just another spore in the wind.
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Suntzu
Geek


Registered: 10/14/99
Posts: 1,396
Last seen: 1 month, 20 days
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Re: Agar...what next? *pics* [Re: Seuss]
#543720 - 02/07/02 12:58 PM (23 years, 17 days ago) |
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Don't have a lot of time now, but here's a water culture pic. . .the growth is only a couple days;
I will never go back to slants after using water cultures, they are simpler to make and according to the references will last longer. If you use slant-type tubes, storage space is the same for both. I think the key is getting as little agar as possible in the suspension; less opportunity for contams and the fungi don't seem to mind. I also find it easier to remove a few drops of the water suspension rather than try to pick out some growth off of a slant [or dig through mineral oil. . .ugh].
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Anno
Experimenter



Registered: 06/17/99
Posts: 24,168
Loc: my room
Last seen: 9 days, 16 hours
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Re: Agar...what next? *pics* [Re: Suntzu]
#543861 - 02/07/02 03:16 PM (23 years, 17 days ago) |
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>according to the references will last longer
The reference:
http://www.mycoculture.org/ftpdir/culture-storage-technique
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