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Reasons for slow colonization( on grain?)


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#1 Severian

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Posted 18 January 2023 - 03:50 PM

So I'm a bit perlexed here- and sorry no photos immediately- but, I've been really in the agar game over the last few months, and have succeeded in cloning from an outdoor mushroom, and (almost) achieving an isolate- but the culture on agar is very bright and super rhizomorphic

However, I went agar to grain on the 11th, and still the mycellium is juuust beginning to climb into the area immediately around the wedge-

At first I didn't shake up the jar, so the wedge was face down on all of the grain, so maybe that amount of air was affecting it somehow?

The hydration shouldn't be an issue, I used Yoshi's no prep tek- with rye- 190 g rye to 200 g water ,PC for 90 mins at 15 psi.

I was expecting much faster growth given the healthy look of the mycellium.

None of the jars show any visible sign of contamination.

Any ideas? Or maybe this is just a slow culture?

#2 DocOct

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Posted 19 January 2023 - 03:57 AM

Sometimes when a mushroom isnt used to a substrate it will be really slow to colonize at first.

What kind of mushroom is it?

Did you have any problems growing it out on agar?
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#3 Severian

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Posted 19 January 2023 - 10:13 AM

No problems on agar, though maybe a tad slow- though I've also been playing with agar mixes , including a no carb black tea mix- possible these specific jars were one of the black tea plates ( coloration was little different from my normal mix so a bit hard to tell- food coloring next time!)- so maybe the swap from zero sugars to sugars has got em a bit confused

I'm growing golden teacher, from a clone I took of a mush fruiting outdoors from a spent-sub pile-

On agar, it appears to have some resistance to green mold and some other contams cuz I've seen it eat through them.

I did the first tissue sample last month, so the culture has been on agar for a month - maybe it needs some time remembering! But rye was the grain originally used for spawn so that memory should be in there somewhere

#4 DocOct

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Posted 19 January 2023 - 01:52 PM

Yeah thats really weird, golden teacher is pretty well domesticated.

Whip up a new batch of grains an try again thats really all you can do if they arent colonizing you current grains, do you have any pics by chance?

And no carb agar sounds like a bold move, cubensis typically like carbs. Is there a reason you opted for this route?

You really cant go wrong with potato dextrose agar its pretty easy to make.

Edited by DocOct, 19 January 2023 - 01:59 PM.


#5 Severian

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Posted 19 January 2023 - 02:44 PM

Was fighting bacteria/yeast for a moment, and the black tea agar is antibacterial- plus the lack of sugar means no food for the baddies- still will culture molds, but less aggressively. Mycellium can feed on the polyphenols- growth is a bit slower but its been a help.

Photos to come later

#6 TVCasualty

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Posted 19 January 2023 - 05:14 PM

My guess would also be that it's probably taking a while to adapt to a new food source, assuming no currently-invisible contamination. If that's the case it will pick up the pace at some point (or else contamination will reveal itself).

 

Do you have several jars going that were inoculated from the same plate?

 

If so then they should all be doing the same thing if it's a matter of re-tooling the digestive enzyme(s). It would be uniquely bizarre if multiple jars were all contaminated with something invisible, but nature has no problem being uniquely bizarre so it can't be ruled out entirely yet.

 

Either way it looks like it's a "wait and see" kind of problem since messing with inoculated jars is generally a bad idea.



#7 Severian

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Posted 19 January 2023 - 06:26 PM

Lol nature has no problem being uniquely bizarre

Ya all from the same plate and now I am seeing contamination- curious though that the slow growth started immediately...


Ahh though I think I'm trying too hard to point the finger at a single 'cause'...

Both 'taking time to change food source'

Thus giving the small amount of contam spores time to get ahold.

#8 Severian

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Posted 21 January 2023 - 11:46 AM

Read in stamets big book

'mycellium grown on the same agar media for an extended period of time can become media dependent- losing their ability to digest other media, thus the reason for additives- yeast, peptone etc'

I would have thought this only occured over 'an extended period of time', but perhaps a month / 5 transfers is considered an extended period.




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