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OfflineKifo
Healer


Registered: 09/20/20
Posts: 9
Last seen: 1 year, 10 months
Agar Issues
    #27088878 - 12/14/20 01:35 PM (3 years, 2 months ago)

Hey guys!

I've had great success with agar but as of recently my plates have been reverting from rhizomorphic to tomentose. For the first 3 months of agar work I was just using LME plates. At the 3 month mark I switched to a new supplier of LME and that's when things started to get confusing. When I sectored my good rhyzo plates, they went tomentose. At first I assumed it was the new LME, so I bought more LME from my original supplier that I had good results with. At the same time, I also bought ingredients to make PDYA plates because I knew I would have to switch up the nutrients every few plates. After making the new LME and PDYA plates, I transferred some good rhyzo mycelium to both of the new LME and PDYA plates. The PDYA plates were a great success but the LME was still reverting the rhyzo to tomentose. I am quite confused because I am using the exact same recipe now that I was when the LME was working very well. Here is my LME and PDYA recipes:

LME
20g Agar
20g LME
1L Filtered Water (Berkey Filtration System)

PDYA
20g Agar
20g Mashed Potato Flakes
14g Dextrose
2g Yeast
1L Filtered Water

I experimented with different water (tap and filtered) and found that the filtered works a little bit better but not great. I'm on a well and my tap water has high calcium and other hard minerals. I assumed the added minerals from the tap water would add more nutrients to the mix and cause the mycelium to not want to grow rhyzo. But the filtered water doesn't make a big difference. My thought process is going to the fact that there are too many nutrients in the LME but it is confusing because it is the same recipe that I had success with in the beginning. Here are some photos of a few plates:

Blue Meanie - PDYA



Blue Meanie - LME



You can see in the LME photo that I have some rhyzo coming off of the end (outer most part of my sector) but the sides and bottom of the sector are going tomentose.


Cambodian - PDYA



Cambodian - LME



The same thing is happening to this Cambo plate. Both the Cambo and Blue Meanie are my strongest genetics and when I transfer them to the LME I get some rhyzo. When I transfer my less aggressive mycelium, I don't get any rhyzo, just all tomentose.


NorMel - LME





I started some new LME plates today and dropped the nutrient amount to see if that makes a difference. The recipe I made today was:

LME
20g Agar
16g LME
1L Filtered Water

I will report back with my findings but if anyone else has run into this issue or has any idea what is going on, I would greatly appreciate some feedback.

Thanks everyone!:smile::heart:


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OfflineHaywire
Wetspot Wizard
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Registered: 12/29/13
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Re: Agar Issues [Re: Kifo]
    #27088916 - 12/14/20 02:06 PM (3 years, 2 months ago)

I checked your recipes and can't find nothing out of the ordinary.
I've been using lME for years now and haven't experienced problems.
Tap water works just fine for me as well.

As to why your cultures are going tomentose, I can't really tell.
But if I were you I wouldn't stress too much and just try them out on some grains.
If you have no issues there or at spawning/fruiting, then I wouldn't stress about it.


--------------------
Ciao mamma, guarda come mi diverto


My grows :mushroom2: Outdoor patches


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OfflineLtLurker
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Registered: 01/03/18
Posts: 7,535
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Re: Agar Issues [Re: Haywire]
    #27088924 - 12/14/20 02:12 PM (3 years, 2 months ago)

Yea man, Tomentose growth is fine if it's clean. Or if you kept transferring you could get the plates more uniform in either pattern. Don't stress it.


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OfflineKifo
Healer


Registered: 09/20/20
Posts: 9
Last seen: 1 year, 10 months
Re: Agar Issues [Re: Haywire]
    #27088926 - 12/14/20 02:13 PM (3 years, 2 months ago)

Thanks for the response. I'll definitely transfer a couple of the tomentose plates to grains and then spawn/fruit to see how that goes.

One other thing I forgot to mention though, is that the tomentose looks weak and sparse. It's very thin sometimes, almost translucent.


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OfflineLtLurker
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Re: Agar Issues [Re: Kifo]
    #27088932 - 12/14/20 02:18 PM (3 years, 2 months ago)

You can always transfer a piece of the rhizo sectors to another plate before committing it all to grains.


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OfflineKifo
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Registered: 09/20/20
Posts: 9
Last seen: 1 year, 10 months
Re: Agar Issues [Re: LtLurker]
    #27088951 - 12/14/20 02:26 PM (3 years, 2 months ago)

Definitely, I always make back ups.


Here's one more example from my Tidalwave plates:


PDYA:




LME:



You can see in the PDYA photo, that the mycelium is nice thick tomentose, but on the LME it looks weak and sparse. The LME sector was taken from the PDYA plate shown.


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OfflineDr.Sparkle
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Registered: 06/03/20
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Last seen: 2 years, 2 months
Re: Agar Issues [Re: Kifo]
    #27088960 - 12/14/20 02:31 PM (3 years, 2 months ago)

Might be multiple strains clamouring on one another on top plate? And a more isolated strain on lower plate?


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OfflineLtLurker
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Re: Agar Issues [Re: Dr.Sparkle]
    #27089022 - 12/14/20 03:08 PM (3 years, 2 months ago)

I see what you're saying op. Go with your gut. It very well could be weaker sectors.

Sparkle: Every plate is multiple strains working together and sectoring out. You'd have to do months of plate work or other special methods to get a single strain.


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OfflineDr.Sparkle
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Registered: 06/03/20
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Re: Agar Issues [Re: LtLurker]
    #27089089 - 12/14/20 03:46 PM (3 years, 2 months ago)

My bad poorly written post. I didn’t mean it was an isolated strain. Was just wondering if it could just be that there were just less strains on the lower plate and more on the upper one.


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OfflineLtLurker
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Re: Agar Issues [Re: Dr.Sparkle]
    #27089099 - 12/14/20 03:51 PM (3 years, 2 months ago)

How it looks has nothing to do with how many strains are in a given sector. Here's Verum's PE isolate project for some examples.
https://www.shroomery.org/forums/showflat.php/Number/25178416


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OfflineKifo
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Registered: 09/20/20
Posts: 9
Last seen: 1 year, 10 months
Re: Agar Issues [Re: Dr.Sparkle]
    #27089103 - 12/14/20 03:52 PM (3 years, 2 months ago)

It could be but I doubt it. If I were to do the same transfer to PDYA, it would look a lot different. I'll do a sample sectoring of one plate to both a LME and PDYA and show the results in a couple weeks


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OfflineDr.Sparkle
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Re: Agar Issues [Re: LtLurker]
    #27089106 - 12/14/20 03:52 PM (3 years, 2 months ago)

Ah okay cheers for the link.


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Offlineverytastycheese
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Registered: 11/11/14
Posts: 101
Last seen: 2 years, 11 months
Re: Agar Issues [Re: Dr.Sparkle]
    #27089230 - 12/14/20 05:30 PM (3 years, 2 months ago)

I make mine the same and I've been advised it may be too nutrient dense. I'm dropping mine 8gLME 10gAGAR 500ml water next round. Rhizo I'm told is it stretching out in search of food.


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OfflineKifo
Healer


Registered: 09/20/20
Posts: 9
Last seen: 1 year, 10 months
Re: Agar Issues [Re: verytastycheese]
    #27089359 - 12/14/20 06:54 PM (3 years, 2 months ago)

Cool, we're on the same boat! I'm made new plates today that are exactly the same recipe you're going to try. I'm just about to fire up the hood and do some transfers, I'll let you know how it goes!


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Offlineverytastycheese
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Registered: 11/11/14
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Re: Agar Issues [Re: Kifo]
    #27099648 - 12/20/20 04:49 PM (3 years, 1 month ago)

Any luck on that growth? I'm a few days behind and my transfers are just starting to show so... :wink:


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OfflineKifo
Healer


Registered: 09/20/20
Posts: 9
Last seen: 1 year, 10 months
Re: Agar Issues [Re: verytastycheese]
    #27107473 - 12/25/20 01:59 PM (3 years, 1 month ago)

Merry Christmas everyone, I've got some findings to share!

On my last posts, I had poured some new LME plates that were less nutrient dense and this was the recipe:

LME
20g Agar
16g LME
1L Filtered Water

I did some transfers to the new LME plates and also to some PDYA plates that I knew would work well. Here are the results:

PDYA




LME




The PDYA plate looks as how I would expect for the Melmak OG mycelium I'm working on. You can see the LME plate did not perform as well. It is very weak and sparse. I made 5 more groups of transfers and they all came out the same, the PDYA outperformed the LME plates.

So I was a little disappointing with the findings because I still had not come to a conclusion as to why my LME plates were weak. I thought dropping the nutrient amount would help me get back to rhizomorphic mycelium but that was not the case. So I decided that I should try a new experiment. I was going to try a completely new recipe and I decided to try BRF (Brown Rice Flour). I poured some new BRF plates with a recipe I found to be very widely used:

BRF
9g Agar
6g BRF
500ml Water

I also decided to try another LME recipe that I made up myself. I knew that my PDYA recipe worked well so I calculated the nutrient amount of every ingredient and it was substantially higher in nutrient content than any other recipe I've ever made. So for the new LME recipe I wanted to try, I decided to up the nutrient content by 50% from the original. Here's the original recipe and new recipe:

Original LME
10g Agar
10g LME
500ml Water


New LMEY
10g Agar
15g LME
1g Nutritional Yeast
500ml Water

Here are the results from the new LMEY and BRF recipes:

BRF




LMEY




As you can see from the photos, the BRF performed alright but the LMEY performed very well! The Cambo plate is very thick rhizomorphic growth and the Avery Albino has a nice thick even tomentose growth (I haven't got it to go rhizo yet).

So this tells me that the mycelium needed MORE nutrients rather than less. In all my research, I had never seen anyone say to up the nutrient content. Everyone always says to use less nutrients, but this experiment proved that wrong. I will now be sticking with my new LMEY recipe and my PDYA recipe for my go to agar plates. I will still do some more experimenting but I feel that I am getting very close to understanding the mycelium I am working with.

I now have an nutrient value equation that I can transfer to any agar recipe to determine the correct amount of nutrients in each recipe. The next experiment I will do is with the BRF agar. I am going to try this recipe next:

BRF
10g Agar
15g BRF
500ml Water

When I get to it, I'll post my findings from that recipe. But for now, thank you so much to those who contributed to this post, this community is amazing!

Have a Merry Christmas and happy New Year!


On a side note, here's a 64g wet Cambodian fruit I harvested a couple weeks ago:



Sorry it's not on a scale for proof :frown:


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OfflineTstone
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Registered: 10/16/18
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Re: Agar Issues [Re: Kifo]
    #27107492 - 12/25/20 02:12 PM (3 years, 1 month ago)

What's weird with my agar work, I find I get better rhizo growth using full strength lme 20g per 1000ml and 2g  nutritional yeast. When I was doing 3/4 or 15g lme only and no yeast, growth was way more tometose. Were as most get more rhizo growth with 10-15g lme, 1000 ml  strength. Still can't figure it out, my myc loves the nutritional yeast.  Others get more rhizo with less nutes, me it's opposite. Won't make a plate without it any more.

I'm also at higher altitude, so perhaps my myc likes full strength nutrition to compromise for less available air. Beats me, but I need full strength to get rhizo at all. I'm at 7000ft


--------------------
Wake now, discover that you are the song that morning brings, but the heart has it's seasons, it's evenings and songs of it's own"


Edited by Tstone (12/25/20 02:24 PM)


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Offlinetiptrippy
The Mechanic
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Registered: 09/09/20
Posts: 1,131
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Re: Agar Issues [Re: Tstone]
    #27107515 - 12/25/20 02:35 PM (3 years, 1 month ago)

This is all very helpful to me. Thank you for your experiments and sharing with us! +5 for you.


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Offlinenogoodnamesleft
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Registered: 09/19/20
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Last seen: 9 days, 20 hours
Re: Agar Issues [Re: tiptrippy]
    #27107545 - 12/25/20 03:02 PM (3 years, 1 month ago)

Thank you very much. I have literally been having the same issue with my Agar cultures and suspected low nutrition to be the culprit.

I have PDA supplies ready to try that blend and I will try a stronger LME recipe as well


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OfflineKifo
Healer


Registered: 09/20/20
Posts: 9
Last seen: 1 year, 10 months
Re: Agar Issues [Re: Kifo]
    #27107643 - 12/25/20 04:47 PM (3 years, 1 month ago)

I'm starting to think that nutritional yeast is a key component in agar recipes. It is very high in protein and fiber, where LME, potato flakes, dextrose and BRF are all high in carbohydrates. I think mycelium doesn't just want 'more' nutrients but rather it favors a more complex array of nutrients.

To go back to my BRF recipe I'm going to try next:

BRF
10g Agar
15g BRF
500ml Water

I think what would work even better is something like this:

BRFY
10g Agar
10g BRF
1g Nutritional Yeast
500ml Water

I'll do an experiment with both of the recipes above and let you guys know my findings :grin:


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