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Agar transfer stored in the refrigerator


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#1 rhapsodyclaymore

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Posted 18 June 2021 - 03:52 AM

Hi everyone,

 

I thought this could use another topic. From my last agar adventure, I made 13 transfers with 12 of which have grown out and are finishing up in spawn jars. I stuck 1 of the transfers in the refrigerator right after the transfer to keep it around for when I want to grow it out.

 

In a couple days, white fuzz started growing on the surface in 1 place and then continued to, rather slowly, grow out. It's been in the refrigerator now for 3-4 weeks and the size of the growth is about as big as a nickel.

 

My question is, I can see just under the white that in the agar layer, it is turning brown-ish. I know it could possibly be contam, but I'm going with no. The 3 original 2oz cups I made transfers from, I kept in a cupboard for a couple weeks just to see what happens. They continued to grow and consume a lot of the agar and the blue agar turned dark and brown as they consumed a ton of it. I also threw them into the refrigerator to slow them down, just because.

 

I can assume, possibly, in my case with the nickel growth that the myc is really cold and cannot really move out from that so fast and is consuming the agar underneath it which is why it has turned a bit brown in color? It would make sense, at least to me. It only turned brown-ish in the last week, but the rest of the agar, including the agar chunk from the transfer, look just fine.

 

I only ask as I haven't seen really any good reference photos of agar transfers in the refrigerator that have been sitting for a while to really judge.

 

13th transfer from the original batch stored in the refrigerator for 3-4 weeks since transfer

1.JPG

 

3x 2oz cups from the first spore syringe trial (used as sources for the 13x no-pour agar plates)

2.JPG


Edited by rhapsodyclaymore, 18 June 2021 - 06:15 AM.


#2 coorsmikey

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Posted 18 June 2021 - 06:58 PM

Is the transfer in the middle of the refer dish? If the nickel sized growth was circling out from the transfer then I would suspect that the consumed blue theory would be likely. I would expect a refrigerated sample to look different that one grown at higher temps, but since the nickel  looks mold that I would see growing on leftovers sitting to long in the fridge and it doesn't seem to be coming from the transfer, I would give up on my optimism. Its not a clear enough photo for me to be sure. All your other transfers along with the refer culture in question seem to be consistently centered, would you expect to see grow radiating out from the transfer even if you happened drop some mycelium in a different spot?


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#3 rhapsodyclaymore

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Posted 19 June 2021 - 03:45 AM

So, here's the other problem I had. I had issues getting the piece of agar off the knife blade or in the center, so there could be a lot of points (or 1) here the chunk of agar had landed before being moved to the center by me.

 

Just to point out again, the fuzz spot (nickel) came up like pure white myc during normal growth before turning brown slowly (I guess due to lack of food). The transfer point where the chunk of agar is was taken from a rizomorphic sample which from my limited experience, takes a while to get set up to grow out. The cotton fuzz myc have always grown quicker.

 

Also in the reference stuff (the 3 cups), some of the myc started not from the center, but from the sides due to also squirting more from the syringe than necessary. So there were multiple growth points that have smoothed out after leaving them in the cupboard for more than a few weeks. That's why they look grown out as they have been searching the sides of the cups for more food and have straightened out.



#4 Oldpunk

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Posted 19 June 2021 - 08:18 AM

I haven't stored any in the fridge yet. Need to get a little mini fridge just for this...actually I kinda spaced out all my samples and they all overgrew during a heat wave we just had. I don't know about the brown coloring. The only ones I've had turn brownish are samples of Tampanensis. And I think it's part of its grow cycle where it starts to grow stones.

I also have a little trouble with getting the sample off the knife. I've been trying to pick it up lightly and cut into the new agar a little. Seems to help get the piece to stick where you want it. I've had a couple grow off to the sides where I dropped a piece as well.

#5 rhapsodyclaymore

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Posted 21 June 2021 - 03:45 PM

So, it's still a little brown, however nothing is moving around. It's like the size has stopped. Given, it grew from pure white fuzz so slowly, it only became a nickel in size after 3-4 weeks. Now, likely closer to 5 weeks. I'm going to go with that it was eating as much as it could from the agar and turned it brown underneath. Maybe in a few weeks to a couple months, I'll warm it up in the room and let it grow out.

 

Still, interesting to see how fridge stalled agar transfers turn out.



#6 Microbe

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Posted 21 June 2021 - 06:33 PM

Is the transfer in the middle of the refer dish? If the nickel sized growth was circling out from the transfer then I would suspect that the consumed blue theory would be likely. I would expect a refrigerated sample to look different that one grown at higher temps, but since the nickel looks mold that I would see growing on leftovers sitting to long in the fridge and it doesn't seem to be coming from the transfer, I would give up on my optimism. Its not a clear enough photo for me to be sure. All your other transfers along with the refer culture in question seem to be consistently centered, would you expect to see grow radiating out from the transfer even if you happened drop some mycelium in a different spot?

Consumed blue theory? This is new to me, that does it mean? I have been working 6 12's so i don't have time to even get on here much.

#7 rhapsodyclaymore

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Posted 22 June 2021 - 01:31 AM

 

Is the transfer in the middle of the refer dish? If the nickel sized growth was circling out from the transfer then I would suspect that the consumed blue theory would be likely. I would expect a refrigerated sample to look different that one grown at higher temps, but since the nickel looks mold that I would see growing on leftovers sitting to long in the fridge and it doesn't seem to be coming from the transfer, I would give up on my optimism. Its not a clear enough photo for me to be sure. All your other transfers along with the refer culture in question seem to be consistently centered, would you expect to see grow radiating out from the transfer even if you happened drop some mycelium in a different spot?

Consumed blue theory? This is new to me, that does it mean? I have been working 6 12's so i don't have time to even get on here much.

 

Reworded - Consumed color theory. Blue is just the food coloring I used. The color gets removed and turns brown underneath after myc has been sitting on the agar for too long.



#8 Microbe

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Posted 24 June 2021 - 07:22 PM

Is the transfer in the middle of the refer dish? If the nickel sized growth was circling out from the transfer then I would suspect that the consumed blue theory would be likely. I would expect a refrigerated sample to look different that one grown at higher temps, but since the nickel looks mold that I would see growing on leftovers sitting to long in the fridge and it doesn't seem to be coming from the transfer, I would give up on my optimism. Its not a clear enough photo for me to be sure. All your other transfers along with the refer culture in question seem to be consistently centered, would you expect to see grow radiating out from the transfer even if you happened drop some mycelium in a different spot?

Consumed blue theory? This is new to me, that does it mean? I have been working 6 12's so i don't have time to even get on here much.
Reworded - Consumed color theory. Blue is just the food coloring I used. The color gets removed and turns brown underneath after myc has been sitting on the agar for too long.
I still have no idea about fuck about shit......what is the 'blue theory' that being tossed around? WTF is it? Cuz i will confirm it or squash that shit.

Yeah the fuck i wont!

Edited by Microbe, 24 June 2021 - 07:23 PM.


#9 rockyfungus

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Posted 25 June 2021 - 08:25 AM

My agar will turn from whatever pigments into the base pigments to clear.

So green=blue + yellow, then clears...

LMflake.JPG
Here's red which is just red pigment and you get chunks of pigment and a clearing.


#10 rhapsodyclaymore

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Posted 28 June 2021 - 08:11 AM

Ok hmm, so possible contam. Which, to be honest, is the only plate out of 3-4 dozen that I've made already. I'll just count that as a positive.






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