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Barnabey
Learner



Registered: 06/19/13
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Spore dilution experience !
#19495841 - 01/30/14 10:43 AM (10 years, 11 months ago) |
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Hi all !
I'll try to be short but I doubt I will be.
I've made a spore dilution from a syringe of mex. jalisco that looked vey very dark. The solution in sterelized distilled water is 1/50 of the original syringe.
I'm very new in all of this and just thought I would experiment while my jars take their time.
My goal is of course to isolate promising features :o
Now i'm not sure where to start, I have Paul Stamet's book and read the chapter on genetics but I'm still confused about different things :
Is the idea by diluting plates to get mycelium that didn't mate yet with another germinated spore and make it meet a good contestant ?
I will read any post you advice me to read that talks about where to go from here.
I understand that with much fewer spores in my plate it will be easier to isolate what I want, I'm just not sure of what I want beside rhyzomorphic looking mycelium and that is even a controversed quality ! :p
I understand having good looking mycelium is absolutely not a garanty of getting good yield or yield at all. So I'm assuming that no matter how nice my agar work is, this has to be fruited before confirmed that I did a good job. I am interested in understanding more about primordia formation on agar if that is an approved way to confirm that the strain will fruit without having to transfer them to grain and fruit them.
Anyway, any info about interesting things to do on agar from spores and spore dilution is welcome ! :p
<= Yes I am not sure of where I'm going with that one
-------------------- My cute contaminants
Edited by Barnabey (01/30/14 10:47 AM)
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hushmush

Registered: 12/30/13
Posts: 165
Last seen: 3 years, 8 months
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Re: Spore dilution experience ! [Re: Barnabey]
#19495920 - 01/30/14 11:03 AM (10 years, 11 months ago) |
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This is a great link: Stro's Cleaning and Isolation on Agar http://www.shroomery.org/forums/showflat.php/Number/18430998
Then just search for more stuff on agar
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Barnabey
Learner



Registered: 06/19/13
Posts: 249
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Last seen: 4 years, 7 months
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Re: Spore dilution experience ! [Re: hushmush]
#19501553 - 01/31/14 02:21 PM (10 years, 11 months ago) |
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thanks ! this help ! but does not cove all my questions there :p
Like, is there an easy way to differenciate the hypha from germinated spores that didn't mate yet from dikaryotic myceliyum ? the way I imagine it it my head when doing a spore dilution would be to select the hypha before it mates and make it mate manually in another dish ?
And my biggest concern after all that nice mycelium selection is what's the fastest way to make sure this is a good fruiting strain ? For example, if I was using shiitake for this, is there a way to get an idea of the fruiting ability without having to incoculate logs and wait years for colonization ? casing or fruiting on bag is the only way ? I recall reading about stimulating pin formation even on agar but I might be hallucinating.
I'm looking for information mostly about spore work on agar and not cloning tek :p
I'm sorry this is messy :p I just have so many interrogations that I do not know where to start, and I'm sure I'm too much of a newbie to be attempting this, but :O When you have a glove box and plenty of petri dishes ready to use you go try out stuff i guess
-------------------- My cute contaminants
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bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Spore dilution experience ! [Re: Barnabey]
#19501573 - 01/31/14 02:24 PM (10 years, 11 months ago) |
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the only thing your eyes will see on the dishes is dikaryotic mycelium, you're going to need a microscope to see the hyphae(primary monokaryotic mycelium) turn into dikaryotic, it happens rather soon after the spores germinate.
you didn't need to dilute your syringe, You can just put a single drop on the agar.
A lot of these questions you can easily find the answer for in the top right of your screen there's a little white box. Or watch the lets grow mushrooms video section on agar, or take a look at the thread the last guy posted.
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Barnabey
Learner



Registered: 06/19/13
Posts: 249
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Last seen: 4 years, 7 months
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Re: Spore dilution experience ! [Re: bodhisatta]
#19501611 - 01/31/14 02:32 PM (10 years, 11 months ago) |
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Thanks for the advices And everyone seems to tell me that I shouldn't have diluted the spores (well I used half the syringe none diluted for other plates and a few jars).
I'm trying to understand more about genetics / sectoring / etc.. Pau Stamet in the mushroom cultivator has a part at the end (page 338 339 340) mentionning spore dilution technique as a "technique that can be used to physically separate spores so that individual dikaryotic cultures can be isolated in one step :o <= so this is just a way to lower the amount of transfers but a pure concentrated solution of spores on agar won't take eternity to isolate a colony ?
-------------------- My cute contaminants
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Barnabey
Learner



Registered: 06/19/13
Posts: 249
Loc:
Last seen: 4 years, 7 months
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Re: Spore dilution experience ! [Re: Barnabey]
#19501627 - 01/31/14 02:35 PM (10 years, 11 months ago) |
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(stamets talks about making a 1:10 solution and a 1:100 solution saying a 1:10 solution should have ABOUT 20-50 colonies per plate and 2 to 5 for the 1:100 solution :o So yeah I made a 1:50 solution on agar to see what happens and not sure where to go from there :p
-------------------- My cute contaminants
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bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Spore dilution experience ! [Re: Barnabey]
#19501654 - 01/31/14 02:41 PM (10 years, 11 months ago) |
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The mushroom cultivator is a great book, but it's also published in many versions, chances are the one you're reading is like 20-30 years old now.
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ghiajake
Myco-Viking


Registered: 01/10/13
Posts: 3,957
Loc: Indiana
Last seen: 2 days, 4 hours
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Re: Spore dilution experience ! [Re: Barnabey]
#19501717 - 01/31/14 02:53 PM (10 years, 11 months ago) |
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I believe growing out your regular mutlispore germinations, fruiting them, then selecting the best specimens of the first fruits to form is the best way to isolate the genetics. So many people go through the whole agar isolation thing without ever knowing what characteristics they are isolating, besides "fastest rhizomorphic growth". RR and others have stated several times that mycelium will switch back and forth from the wispy and rhizo growths due to amount of available nutrition. The fast rhizos are lacking nutrients and are branching off to find nutrients. The wispy growth has plenty of nutrients and doesn't need to go searching for it. Cloning the best and fastest fruits will give you the best isolation in just one step, instead of continually selecting genetics you have no idea how they'll fruit. Hope that helps some.
Edited by ghiajake (01/31/14 02:54 PM)
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bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Spore dilution experience ! [Re: ghiajake]
#19501731 - 01/31/14 02:57 PM (10 years, 11 months ago) |
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I think maybe you should start with a PF tek grow or something first just to see what you're working with before jumping right into agar. I do applaud you for doing so but it seems like you have a lot of research to do. Not to be a downer but I'm really just trying to help see to it that you stay in this hobby and don't get deterred by failure.
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hushmush

Registered: 12/30/13
Posts: 165
Last seen: 3 years, 8 months
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Re: Spore dilution experience ! [Re: bodhisatta]
#19506331 - 02/01/14 02:02 PM (10 years, 11 months ago) |
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I think to start on agar is good to make sure you have clean contaim free mycelium to start transfers with. You don't need an isolation. You can have a plate with 20 cultures in it and transfer with that. When you start isolating you will have to fruit each to see its traits.
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