Paradox
©
Fisana

Jump to content


Photo
- - - - -

Substrate PH, Agar, LC and other projects


  • Please log in to reply
22 replies to this topic

#1 FunnyFarmer

FunnyFarmer

    Mycotopiate

  • Free Member
  • 324 posts

Posted 16 November 2021 - 03:53 PM

As some of you know I've had one failure after another the whole summer, actually starting in mid spring. I think I found the reason, I found out that the PH was too low when I mixed the sub for my latest grow. Seeing as how this is the first time I've checked it I gotta wonder how far off the previous grows were. This one started around 6 - 6.3 so I added gypsum to bring it up to 7, checking several areas for hot spots before putting it into the oven for pasteurization.

 

I've got high hopes for this one, I'm spawning 3 qts of GWM to it. It will be a little over a 2:1 sub to spawn ratio using a manure based sub. I'll check the PH again before spawning it just to be sure. The quarts will wait in the meantime if it doesn't check out. If it goes too basic what would you recommend to correct it?

 

The present grow has been a disappointment, genetics worked against me. It is Treasure Coast and for the most part they average about 2" tall with a spindly stem and less than 1/2" diameter caps. The sub they pull with them probably weighs more then them. It would probably take 100 of them to equal one KSSS shroom for size and weight.

 

The experiments with agar and LC are progressing nicely with 4 LC jars and another 5 agar varieties that I recently did A2A transfers. 4 of the agars have recovered nicely but one with Ecuador doesn't look promising. I'm experimenting with a yeast based agar that I came across on Youtube and so far the results are promising. Getting great rhizome growth from the first varieties that are now in the fridge waiting for their turn at LC. One of the other recent transfers is PB and it's really responding to the agar compared to the others, Argentina, GT and

 

Another question, how much mycelia do I allow to settle in the jar before putting it in the fridge? Right now I've got about 1/2" of mycelia in the bottom of a pint jar that was mixed at 4% corn syrup and doesn't seem to be growing any more. When I put them on the stir plate there's a good cloud and the water is clear after they've settled.



#2 jkdeth

jkdeth

    Mycotopiate

  • Free Member
  • 2,584 posts

Awards Bar:

Posted 17 November 2021 - 12:46 PM

On that ph, Im not sure you're getting an accurate reading. Especially if it seems gypsum brought it up. Gypsum doesn't directly affect ph.

An acidic can be more contaminate prone though . test the the water you use to hydrate. I know of of a couple people who had problems and found it was due to acidic tap water.. You can lime the water up to about8 plus, prior to hydration.
  • bezevo and Salty117 like this

#3 FunnyFarmer

FunnyFarmer

    Mycotopiate

  • Free Member
  • 324 posts

Posted 17 November 2021 - 10:19 PM

I was perusing another thread here and he recommended using gypsum to adjust from an acidic condition. Was I steered wrong?


Edited by FunnyFarmer, 17 November 2021 - 10:51 PM.


#4 jkdeth

jkdeth

    Mycotopiate

  • Free Member
  • 2,584 posts

Awards Bar:

Posted 18 November 2021 - 10:38 AM

I don't know when people started that bit about gypsum. Gypsum has been used a long time, but Iys lime you want to treat an acidic conditions.

Gypsum was used as a ph buffer. Substrates tend to acidify over time, gypsum helps slow that process.
  • Myc likes this

#5 Myc

Myc

    El Jardinero

  • App Administrator
  • 7,808 posts

Awards Bar:

Posted 18 November 2021 - 10:47 AM

You may have read a comment where I referred to gypsum as a pH-up buffer. Maybe that was confusing. It is a buffering agent that is added to a substrate with the intent of balancing out acids as they are produced in the growth cycle of fungi.

 

You mention a "yeast based agar". Is this YPD?

Just curious to see what you're working with. Could you detail please?



#6 FunnyFarmer

FunnyFarmer

    Mycotopiate

  • Free Member
  • 324 posts

Posted 18 November 2021 - 01:45 PM

Myc, on 18 Nov 2021 - 10:47 AM, said:

You may have read a comment where I referred to gypsum as a pH-up buffer. Maybe that was confusing. It is a buffering agent that is added to a substrate with the intent of balancing out acids as they are produced in the growth cycle of fungi.

You mention a "yeast based agar". Is this YPD?
Just curious to see what you're working with. Could you detail please?

The yeast agar project started with this video I came across on Youtube

I decided to give it a try and have been impressed with the results compared to the straight LME based agar.

DSC01773 (Medium).JPG DSC01774 (Medium).JPG DSC01775 (Medium).JPG DSC01776 (Medium).JPG

Basically all they are doing is adding a small amount of nutritional yeast to the recipe. I found out I needed to cut back a little on the agar cause it was getting too viscus as it was coming up to temp. These are all at least 2nd gen samples and at most 3rd gen grows. I added activated charcoal to the mix shortly before it comes to a simmer. This is the recipe I use for a 200ml batch. Upscale as needed. All the above plates are residing in the fridge for the time being. The chop taken out of some was for LC and didn't see any reason to pitch them afterwards although I could use them to start some pint rye jars for future G2G projects...

3.75g telephone agar
3g LME powder
.1g Nutritional yeast (Bobs Red Mill Organic)
.8g charcoal or about 1 tsp (?)

I wait until the agar gets close to 180 F before adding the charcoal as it can obscure the bubbles forming on the bottom of the beaker. I've got one of those heated stir plates, very handy. I do the so called no pour method (kinda misleading) and leave the lid off the plates to let most of the moisture escape. Yea, doesn't sound like a good idea but I've done several pours in the kitchen without any kind of sterile procedures and have only had a very few contams get past the PC process. My first attempt at agar work I tried the pour process in a SAB and ended up tossing at least 90% of the dishes. I'll never go back to that process.

Lately I went back to my prints and made agar plates from them. For the most part I got good germination from all but one. I did a A2A transfer from them and all have responded to differing degrees with the PB leading the pack and KSSS following close behind with Argentina, GT and then Ecuador in trailing position. An agar plate I did from a syringe failed so I tossed that one. As fast as these are progressing I'll need to make another batch of plates before too soon.

As I'm still somewhat of a noob I figured I was behind the curve when it came to this tek and didn't bring it to anyone's attention here. Do I need to start a new thread?

BTW it wasn't you I found the info from and it was an older post, iatebadshrooms was the author in 2013.


Edited by FunnyFarmer, 18 November 2021 - 03:20 PM.

  • Myc and coorsmikey like this

#7 FunnyFarmer

FunnyFarmer

    Mycotopiate

  • Free Member
  • 324 posts

Posted 18 November 2021 - 10:37 PM

So I went to check on my LC and out of the corner of my eye I spotted some white in the GWM tub. Upon closer examination instead of a white reflection off the tub I've got some impressive tufts of mycelia coming through already, the day after spawning it. In the past the soonest I've had mycelia after spawning is three days. If I can avoid contamination and everything goes well this could be an awesome grow. I need to let you guys know that this isn't a product of the yeast agar experiment, I started those jars way before that. But that being said I wonder how well that GWM dish in the previous post would respond...


  • Salty117 likes this

#8 FunnyFarmer

FunnyFarmer

    Mycotopiate

  • Free Member
  • 324 posts

Posted 19 November 2021 - 08:55 AM

Apparently not all varieties are comfortable with the yeast agar. As you can see from the previous post every one of those is quite content growing on it but of the newest batch GT and Ecuador don't want to go much beyond the sample while the others are fine with it. Every initial sample was taken from the healthiest area of the original dish. I may need to take some more samples from the initial dish or start another from scratch to verify their behavior first. I seem to get a better start from a sterile swab instead of scraping spores from the print, don't know why that would be.
  • jkdeth likes this

#9 rockyfungus

rockyfungus

    Mycotopiate

  • Free Member
  • 1,844 posts

Awards Bar:

Posted 19 November 2021 - 10:32 AM

Never had issues with nutritional yeast.

Myc seems happiest when it moves from nutrition source to nutrition source.

I usually do straight grain water. Then MEA, then a combo with yeast.
If I keep using the same then I do some manure agar, etc.
  • Myc likes this

#10 FunnyFarmer

FunnyFarmer

    Mycotopiate

  • Free Member
  • 324 posts

Posted 19 November 2021 - 12:42 PM

I made a batch of agar dishes and got the agar amount wrong, instead of 3.85 g go with 4 g. They had the consistency of diarrhea and wouldn't solidify to any extant whatsoever.

 

Manure agar? Never came across that one, until now. So whats the story/details with that?


Edited by FunnyFarmer, 19 November 2021 - 02:12 PM.


#11 rockyfungus

rockyfungus

    Mycotopiate

  • Free Member
  • 1,844 posts

Awards Bar:

Posted 19 November 2021 - 01:53 PM

Any agar recipe throw a turd in it.
Like take a normal agar recipe 500ml H2O, 10g MEA, 10g agar.

Take a small piece of manure shred it up in that 500 ml H2O let it turn to a brownish tea. Strain out the actual poop. Now you got all the goodies for manure helping your mushrooms get the shit they actually like. No real recipe but that's how I experiment.

Also use wood teas, compost teas, any nutrient source really.


Edited by rockyfungus, 19 November 2021 - 01:57 PM.


#12 FunnyFarmer

FunnyFarmer

    Mycotopiate

  • Free Member
  • 324 posts

Posted 20 November 2021 - 09:57 AM

I checked the GWM grow this morning and was astonished to see what looks to be a pin, 3rd day after spawning it! Is GWM known for this kind of growth rate? This is unreal in my short experience, a few months over a year, with growing shrooms and outside everything I've read about shrooms. My last grow (TC) took a little over two weeks before it pinned (normal) but everything about it has been pathetic, tiny shrooms hardly worth keeping and seems to have quit after one flush. I'm ignoring it for the time being and may bury it in the shroom garden when the GWM comes of age.
  • Salty117 likes this

#13 Salty117

Salty117

    Mycotopiate

  • VIP
  • 429 posts

Posted 20 November 2021 - 05:00 PM

If I remember correctly, GWM is an Albino A+ isolate that produces particularly thick fruits. I have some GWM swimming in LC that I just got out of the fridge a couple days ago to start putting to grain or BRF cakes (undecided which route I'm going for them). I'm not familiar with the cultivation of AA+, so I'm not sure how long is "normal" for them or GWM to reach pinning from introduction to fruiting conditions. When I get to the point of spawning to bulk, I'll try to keep note of how long it is for them to start to form pins.


Edited by Salty117, 20 November 2021 - 05:00 PM.


#14 Myc

Myc

    El Jardinero

  • App Administrator
  • 7,808 posts

Awards Bar:

Posted 20 November 2021 - 06:04 PM

Thanks for the video embed.

They're making what I use as a yeast propagation media. LOL

My recipe is not different in any way - measurements and proportions are the same. I always suspected that higher fungi would enjoy the recipe.

Thank you so much for taking the time to confirm my suspicions.

 

You probably saved me a month of experimentation - Thanks Big Time!!!


Edited by Myc, 20 November 2021 - 06:08 PM.

  • FunnyFarmer likes this

#15 rockyfungus

rockyfungus

    Mycotopiate

  • Free Member
  • 1,844 posts

Awards Bar:

Posted 20 November 2021 - 06:31 PM

Yeast added to either LC or agar is pretty standard at this point. Dropping the nutrients (Mea) and filling the rest in with nutritional yeast gives rhizo for a lot of people.

I don't see a remarkable difference personally. Varying it up is the key to working with a culture long term IME. 


  • Myc likes this

#16 FunnyFarmer

FunnyFarmer

    Mycotopiate

  • Free Member
  • 324 posts

Posted 20 November 2021 - 09:53 PM

If I remember correctly, GWM is an Albino A+ isolate that produces particularly thick fruits. I have some GWM swimming in LC that I just got out of the fridge a couple days ago to start putting to grain or BRF cakes (undecided which route I'm going for them). I'm not familiar with the cultivation of AA+, so I'm not sure how long is "normal" for them or GWM to reach pinning from introduction to fruiting conditions. When I get to the point of spawning to bulk, I'll try to keep note of how long it is for them to start to form pins.

If I'm not mistaken (and I could be) the AA+ and GWM (?) are in the PE family and needs a verm casing on top to stimulate fruiting? Oh, never mind, I see you don't have experience with these cultivars so I guess this is a question for the general community. At the rate this is colonizing it'll be totally surface colonized by tomorrow. This is unreal for me and I don't want to fuck it up, especially after so many failures in a row during the summer. I'm pretty sure now that they all had PH too far to the acid side that attracted trich. One other thing I did was oven pasteurize the sub, manure based, instead of pillow case in hot water. The oven did cause me some concern when it came awfully close to 180 for much of it's time in there. I just hope the thermometer was erring on the high side...
 



#17 Salty117

Salty117

    Mycotopiate

  • VIP
  • 429 posts

Posted 20 November 2021 - 10:21 PM

All that I have been able to find is that it's an albino variant of the A+ specimen, but that's about as far as my research has gotten me. I haven't found anything on the source of who "isolated" that A+ variety, or what really makes it unique as a specimen of the cubensis genus.

According to SacredMeds(dot)com (take their description with a grain of salt if you want) their description of Great White Monster is..

 

 

 

Great White Monster Cubensis

Great White Monster Cubensis Psilocybin are an albino Magic Mushroom. They get their name from the unique rusty white spores they produce. Great for the outdoors, this euphoric strain really intensifies sensation and brightens vision. Expect higher than average potency with the Great White Monster.

This strain was originally a cross between Puerto Rican and the A-strain. However, the Great White Monster Cubensis was born out of accidental exposure to quaternary reactive ammonium siloxane. This is a special strain as it is the only P. Cubensis strain known to have both albino and leucistic spores. The fruiting bodies of this magic mushroom are white like albino shrooms but it is in fact leucistic (partial loss of colour pigmentation). The GWM is also known to have large fruits, slow to grow but prolific grower.


Edited by Salty117, 20 November 2021 - 10:24 PM.


#18 FunnyFarmer

FunnyFarmer

    Mycotopiate

  • Free Member
  • 324 posts

Posted 20 November 2021 - 11:20 PM

That write up sounds familiar, like from another source I came across. Slow to grow, huh? That doesn't sound like mine...  As for the rest of it it's a matter of time before I find out. I guess I'll need to get a tissue culture and clone it for future microscopy research. Then I can put another tick mark on my bucket list of shroomology.



#19 Salty117

Salty117

    Mycotopiate

  • VIP
  • 429 posts

Posted 21 November 2021 - 12:16 AM

Yeah, I mean it's also some online canadian vendor so that's why I said to take it with a grain of salt if you want to. Information surrounding isolate lineages can be difficult to come by so it's hard to verify one source against another in many cases, it is nice to know though that it rings a bell to something you've previously read.

I noticed that too about the mention of the growth rate, but I wouldn't take that as a sign that it's not what you have. Time will tell  :meditate:



#20 Salty117

Salty117

    Mycotopiate

  • VIP
  • 429 posts

Posted 21 November 2021 - 01:18 AM

Also, sorry to double post here but I keep my eyes out for "alkaline water" like this for being on sale and use this in combination with pulverized oyster shells to raise the Ph of my substrate.

 

WTR.jpg






Like Mycotopia? Become a member today!