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OfflineBluePapayaSky
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Registered: 06/27/16
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Agaricus Bisporus on petri dish
    #23386328 - 06/27/16 01:31 AM (7 years, 8 months ago)

This is my first try to obtain mycelium

i'm trying to clone agaricus bisporus (from the market) into Petri Dish

25/6 - i made PDYA plates and transferred 3 pieces of mashroom tissue to the center of 3 plates

27/6 Today -
one plate shows no growth but 2 white dots of bacteria
one plate shows little odd shaped growth of transparent mycelium from the center
the last plate shows very nice growth of transparent mycelium (hard to see on the light colored PDYA)

My question is - why transparent? is it ok?

26/6 - i made PDYAH (horse manure) plates (this time dark brown) and transferred pieces of mashroom tissue to 5 plates this time 3 pieces in each plate


i'll try to update
sorry for the lack of images my camera is very bad

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Invisibleweetsie
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Re: Agaricus Bisporus on petri dish [Re: BluePapayaSky]
    #23386656 - 06/27/16 05:49 AM (7 years, 8 months ago)

When you only have a tiny bit of mycelium, as you would expect 2 days after cloning it appears transparent. In a couple more days as it thickens up it should look more white and opaque.


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OfflineSpinScratch
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Re: Agaricus Bisporus on petri dish [Re: BluePapayaSky]
    #23386674 - 06/27/16 06:03 AM (7 years, 8 months ago)

I just tried this recently. I made 3 plates of a store bought clone portobello. 2 plates had no growth. 1 plate grew about 1-2 cm of thin, wispy mycelium and then stalled out and turned brownish. Then 2 weeks later it came back to life and grew another half cm and stalled out again and turned brown.... gave up and ordered and LC syringe last night.

What substrate do you plan to grow it on?

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OfflineBluePapayaSky
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Re: Agaricus Bisporus on petri dish [Re: SpinScratch]
    #23387635 - 06/27/16 12:44 PM (7 years, 8 months ago)

weetsie thats greate news

SpinScratch i did mine with white buttons
i hope it wouldn't get to that for me
my mission is to start a cloture with store bought mushroom
one crop is all i need for spores to my heart's contempt (:

27/6 Today - the PDYAH plates shows some nice growth
but those white dots :mad2: (oddly in the color of the mycelium)
one of them got to my good PDYA plate
and to one of the PDYAH
i flamed my knife after every transfer
do you think its because all my plates seats in the same box without parafilm seals?

Edited by BluePapayaSky (06/27/16 12:58 PM)

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Invisibleweetsie
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Re: Agaricus Bisporus on petri dish [Re: BluePapayaSky]
    #23388311 - 06/27/16 04:49 PM (7 years, 8 months ago)

If you don't have parafilm you can use plastic wrap or micropore tape but definately wrap your plates with something.

Once you get a clean culture from a clone id hold onto it and use it for future grows. Starting from spores is unlikely to ever produce results that will match professional strains made for the industry.


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OfflineBluePapayaSky
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Re: Agaricus Bisporus on petri dish [Re: weetsie]
    #23389950 - 06/28/16 04:13 AM (7 years, 8 months ago)

weetsie i covered the plates with plastic wrap and it feels much better
but should i poke some holes in it?

by the way i transferred a piece of mycelium from that good PDYA plate to a new PDYAH let's see how it goes

someone know's what it means when the mycelium turns a bit brownish ?

Edited by BluePapayaSky (06/28/16 04:17 AM)

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Invisibleweetsie
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Re: Agaricus Bisporus on petri dish [Re: BluePapayaSky]
    #23390018 - 06/28/16 05:20 AM (7 years, 8 months ago)

I'm not speaking from experience as Ive never used plastic wrap but I've read it allows for sufficient air exchange so no need for holes.

When I cloned a portabello the first thing that happened was a brown cloud appeared under the the tissue followed by white mycelium along the surface of the agar. When I broke up the agar the brown stuff appeared to have mycelium running through it which makes me think that's just what bisporus does. Transfer it to grain and see what happens.


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OfflineBluePapayaSky
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Re: Agaricus Bisporus on petri dish [Re: weetsie]
    #23390207 - 06/28/16 07:24 AM (7 years, 8 months ago)

thanks weetsie
when ill have a sufficient enough plate i thought id make liquid culture first and only then grain it
will it work well to just cut pieces from the plate and transfer directly to grain?

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Invisibleweetsie
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Re: Agaricus Bisporus on petri dish [Re: BluePapayaSky]
    #23390926 - 06/28/16 12:07 PM (7 years, 7 months ago)

Agar to grain is as close to the "proper" way of doing things as you will get.
Liquid culture works and might be a little quicker initially but I don't think it's worth the extra chance of picking up a contam.
Once you have your initial colonised jar you can expand it with with grain to grain very quickly.


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OfflineSpinScratch
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Re: Agaricus Bisporus on petri dish [Re: weetsie]
    #23391344 - 06/28/16 03:10 PM (7 years, 7 months ago)

Agar to grain is a much safer bet. I just ordered a portobello LC, but I'll be putting it on agar before I do anything with it.

And yes, cut a small piece of agar from the dish and quickly and efficiently put it inside the grain jar.

Edited by SpinScratch (06/28/16 03:11 PM)

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