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Newbie to Agar - Growth Happening, What Now?


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#1 PaceArrow

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Posted 01 June 2021 - 04:42 PM

Ten days ago I made agar with activated charcoal added, and the next day inoculated several agar cups with spore syringes and another two cups with clone tissue. Here are the results so far, and I am hoping for input on how things look and what to do next. Please note that all out of focus blob things in the images are water condensation on the lid of the cup. No contaminants present, as far as I can see.

 

The recipe I used is 8.25g agar agar + 8.25g LME + 2g activated charcoal in 550ml distilled water. 

 

As always, THANK YOU!

 

Mazatapec spore syringe: 

 

IMG_1899.jpeg    IMG_1903.jpeg

 

Panaeolus Bisporus spore syringe: 

 

IMG_1904.jpeg    IMG_1905.jpeg

 

Golden Teacher tissue samples:

 

IMG_1906.jpeg    IMG_1909.jpeg


Edited by PaceArrow, 01 June 2021 - 07:49 PM.

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#2 Sidestreet

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Posted 01 June 2021 - 06:08 PM

Your growth looks happy and whispy.  What agar recipe are you using?

 

You could make liquid cultures or take wedges to grain jars to fruit...

 

Long-term, rather than try to isolate strains on agar, you could try to keep it diverse and do some test grows so you can pick out some good fruits to clone.  Then once you've tested out your clone candidates and know you have good genetics, you could work toward preserving them by taking them to multiple slants or generously-poured agar plates and putting them in the fridge for storage and to use whenever you like.


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#3 PaceArrow

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Posted 01 June 2021 - 07:28 PM

Your growth looks happy and whispy.  What agar recipe are you using?

 

You could make liquid cultures or take wedges to grain jars to fruit...

 

Long-term, rather than try to isolate strains on agar, you could try to keep it diverse and do some test grows so you can pick out some good fruits to clone.  Then once you've tested out your clone candidates and know you have good genetics, you could work toward preserving them by taking them to multiple slants or generously-poured agar plates and putting them in the fridge for storage and to use whenever you like.

 

Thanks very much, Sidestreet. The recipe I used is 8.25g agar agar + 8.25g LME + 2g activated charcoal in 550ml distilled water.

 

The Golden Teacher clone mycelium is from the tissue of a very nice fruit so I think I will opt to make a liquid culture from it. I like the idea of test grows for the others, and will wait for the cups to be more fully colonized and then will cut the agar and add to some grain jars. I was a bit intimidated by the whole agar thing before actually doing it, and now that I have it's not scary at all, especially since I have you kind and knowledgeable folks to back me up!


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#4 Sidestreet

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Posted 02 June 2021 - 04:12 AM

 

8.25g agar agar + 8.25g LME + 2g activated charcoal in 550ml distilled water.

 

That's cool, the growth is a little bit thinner than I'm used to seeing, but your recipe stretched the nutrients a little bit thinner than mine (10g agar + 10g LME + 500 mL water) so that could be the reason.  The thinner growth is not necessarily a bad thing at all.

 

Glad you dove in to agar.  I held off for a long time because I assumed I couldn't do it in a glovebox. 



#5 Arathu

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Posted 02 June 2021 - 04:31 AM

Pace you're opening up an entirely new world mycology for yourself.........

 

For example, from this culture I would personally make some transfers to new agar from the rhizome looking aggressive mycelium as indicated by the red circles......transfer just the tips

 

I've had pretty good results from grabbing and growing out fungus with these characteristics.......NOT always but that of course is all part of the fun....

 

post-113856-0-96440400-1622625960.jpeg

 

Nice work man! You're definitely getting there now........

 

Grow out new plates, use wedges to inoculate grain jars and expand from there......there are all kinds of possibilities from this point......

 

Which IS a big part of the point of culturing on agar........

 

A

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#6 Oldpunk

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Posted 02 June 2021 - 07:48 AM

Sweet! Those look good for a first try.
(better than mine did)


Isn't it fun when they start growing? After you transfer the little spots like Arathu pointed out they will grow more evenly.
You're pics are pretty clear. What did you use for containers?

#7 FunnyFarmer

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Posted 02 June 2021 - 09:48 AM

I've got all I need to do agar too but am also somewhat intimidated. I'll get past it as I've got a couple prints that need to be put to sub. One is tidal wave that I got from a vendor and the other gmw (?) that I got from a former forum member in exchange for a pink buffalo print. He got the better end of the deal, a thick coal black print while all I got was a wispy smeared print in return. It'll take some work to get a good agar sample from it. The TW needs some work as well as it wasn't a good print either, I tried making a syringe from it but it contamed almost immediately. Oh well...

#8 PaceArrow

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Posted 02 June 2021 - 01:39 PM

Arathu thanks very much for the input! So when one is cloning from a mushroom fruit with the desired traits, one can still refine those traits by choosing and culturing the more adventurous rhizomic arms in agar?

 

Oldpunk, the 4-oz portion cups I used are these, which are PP5 and thus able to handle PC heat: https://www.amazon.c...0?ie=UTF8&psc=1

 

I followed Josex' tek for modifying the cups with a hot screw driver blade and micropore tape: https://www.shroomer...Number/25137693 (Hope its okay to post links from the other site). I also used the recipe from his tek, though in retrospect next time I will add slightly more LME since I am also adding charcoal. 


Edited by PaceArrow, 02 June 2021 - 02:26 PM.

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#9 PaceArrow

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Posted 02 June 2021 - 02:30 PM

 

 

8.25g agar agar + 8.25g LME + 2g activated charcoal in 550ml distilled water.

 

That's cool, the growth is a little bit thinner than I'm used to seeing, but your recipe stretched the nutrients a little bit thinner than mine (10g agar + 10g LME + 500 mL water) so that could be the reason.  The thinner growth is not necessarily a bad thing at all.

 

Glad you dove in to agar.  I held off for a long time because I assumed I couldn't do it in a glovebox. 

 

 

I thought the growth might be a bit thin also, especially on the P. Bisporus, so next time I will up the LME slightly. I dont have a glovebox but I do have my modified SAB which has an ozone generator attached to it, which I think helped keep the bad critters away from the agar..



#10 PaceArrow

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Posted 03 June 2021 - 01:16 AM

The Panaeolus Bisporus mycelium on the agar is very fine and not at all dense. Will this still work if I cut agar wedges and put in a grain jar, or will the fine mycelium not be able to handle the disruption?

 

IMG_1904.jpeg


Edited by PaceArrow, 03 June 2021 - 01:17 AM.

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#11 Arathu

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Posted 03 June 2021 - 05:11 AM

Pace....

 

Yes, even with clones the expressions are there, and it still amazes me to see those characteristics when the mycelium is running our substrates........very cool stuff.

 

As a side note, honey and potato flakes (instant potatoes), even small amount of dried manure, not to mention MANY other additives, can be added to your base agar mix to see how the fungus reacts. Make a batch of agar with a small amount of of bone dry and powdered HPOO and put some of that P. bisporus transfers to it. I look for aggressive, healthy, growth. Also keep in mind that NOTES are imperative to keep track of what you're learning about each species. Obviously each species will have it own unique characteristics......

 

Keep going, you're into the really cool stuff now.....

 

A


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#12 PaceArrow

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Posted 03 June 2021 - 12:10 PM

Pace....

 

Yes, even with clones the expressions are there, and it still amazes me to see those characteristics when the mycelium is running our substrates........very cool stuff.

 

As a side note, honey and potato flakes (instant potatoes), even small amount of dried manure, not to mention MANY other additives, can be added to your base agar mix to see how the fungus reacts. Make a batch of agar with a small amount of of bone dry and powdered HPOO and put some of that P. bisporus transfers to it. I look for aggressive, healthy, growth. Also keep in mind that NOTES are imperative to keep track of what you're learning about each species. Obviously each species will have it own unique characteristics......

 

Keep going, you're into the really cool stuff now.....

 

A

 

Thanks again, Arathu. Your information is very appreciated! 


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#13 rockyfungus

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Posted 03 June 2021 - 08:21 PM

The less nutrients the faster it seems the myc starts searching for places to expand.

With higher nutrients sometimes a plate can almost stall out and take it's time gorging itself on all the extra food. 


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