After more than 2 years of sitting in spore water, the Redboy lives!
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Posted 17 April 2021 - 05:29 AM
Two PE liquid cultures, one with only the most rhizo mycelium and the other with a bit of a mix.
I used 4 grams of LME for 100 mL of water in each. PCd for 25 minutes at 15 psi.
Because of the color of the broth, I think I'll stick with karo for LCs from now on. It comes out nice and clear.
Side note, another one of my PE plates is finally showing growth. I must have gotten the tiniest bit of mycelium in there when I transferred. So four out of five were good!
Edited by Sidestreet, 17 April 2021 - 05:31 AM.
Posted 17 April 2021 - 08:14 AM
Is that normally how you make your LC? I use < .2 g / 100 ml water. My recipe is .6 g LME and 350 ml water in a pint jar. PC for 15 mins.
Posted 18 April 2021 - 03:50 PM
Nope, I've never used LME in a liquid culture. I was going for 4% sugar content.
Posted 19 April 2021 - 04:55 PM
Yeah, there's some gnarly sediment in there. So, not 4% sugar when using LME?
I'll do new ones and just use honey. I know honey.
Posted 19 April 2021 - 06:51 PM
I'm using 1% for lme lc. JRH recipe is 0.2%
4% is too much sediment and it seems to grow slower than lower percent.
Posted 19 April 2021 - 07:43 PM
Posted 24 April 2021 - 10:01 AM
Okie dokie, I did new LCs with 1% LME. The old ones that had 4% were growing some very small bits of mycelium but it seemed to be on the surface of the liquid.
These look much better, though there's still a little color to them.
I put a wedge of each of my four good Penis Envy plates into both of those LCs. I want to have multiple strains at this point, and I'll select from the best performers. The downside is that doing more transfers to each LC increased the chances of contamination.
Here are the plates I used. I'll give them a day or two to recover and them move them to cold storage:
Also, my two Redboy test jars are getting close to being done. I'm going to let them consolidate and them put them in the fridge too, so the other Redboy jars I started a couple of days ago can catch up.
Right now the goal is to clone from both varieties, do a test grow of the clones, and make culture slants for storage. By that time I'll probably have plenty of fruits to make the honey I want. Also I'll get some spore prints and swabs.
Still nothing from the 2017 varieties. I see tiny spots of bacteria starting on one of the plates, so if the test jar doesn't germinate it may be all over for that variety.
Edited by Sidestreet, 24 April 2021 - 10:11 AM.
Posted 24 April 2021 - 10:15 AM
I'm going to run out of likes before noon......
Posted 24 April 2021 - 05:32 PM
Posted 24 April 2021 - 07:16 PM
I wonder if that last plate is a single strain. It looks so uniform. That's the plate where I got the tiniest little speck of mycelium into the plate and it didn't show any growth for days. I thought nothing was going to happen at first.
Edited by Sidestreet, 24 April 2021 - 07:16 PM.
Posted 26 April 2021 - 05:17 PM
Since I've got four good PE plates in my pocket, I decided I could retire their daddy and get a couple of open-lid pics. It's got contamination hiding under all that good stuff.
Edited by Sidestreet, 26 April 2021 - 05:19 PM.
Posted 26 April 2021 - 11:24 PM
Posted 27 April 2021 - 03:44 AM
There was a pink contaminant that was growing next to the mycelium, but then the mycelium grew over it. I could still see it by looking at a light through the plate.
Posted 27 April 2021 - 07:08 PM
PE did really well for me off of MSS (t1 off swab). The more I isolated it the less of a canopy I had.
I used to use agar like yourself. Now I just take from the earliest clean plate. Seems to give you the most to select from instead of just saying this agar looks nice and uniform.
Nice and uniform may not fruit...
Edited by rockyfungus, 27 April 2021 - 09:38 PM.
Posted 28 April 2021 - 04:08 AM
That's what I've heard more recently--just because it's uniform and rhizo doesn't mean it's a strong fruiter, just that there's a single strain on the plate. So, instead of isolating several strains and growing them all out to see if any are good performers like I used to see people do, it seems better to grow out a bunch of strains (multispore or non-isolated mycelium on agar) and take a clone. Then you know you have the good performer.
For this, I was just going for clean plates up until now. Once I had them, I still wanted to have that mix of genetics so I've put multiple wedges into each LC. I'll grow a few cakes out of those LCs to find a good clone candidate to take back to agar.
Update on the LCs: They're growing decent mycelium, but 1g of LME per 100 mL of water still resulted in more sediment than I would like to see. I'll definitely adjust the LME downward again for LCs if I keep using it.
Edited by Sidestreet, 28 April 2021 - 04:16 AM.
Posted 28 April 2021 - 04:25 AM
I have mixed feelings about the sediment from LME, it seems messy in the beginning but as the myc grows out it seems to gobble it up cleaning out the jar. I'm still using it, though aesthetically the LC's I made with just Corn syrup look nicer.
Posted 28 April 2021 - 07:00 AM
I don't mind malt extract for LCs, the sediment seems to allow the myc to get an early scaffold. It eventually clears up pretty nicely. I may be in the minority but I even like a grain water LC.
Posted 29 April 2021 - 04:31 AM
Starting test jars for my PE LCs. When I make cakes I always seem to have extra, but it's so easy to just remember 1/2 cup verm, 1/4 cup BRF, 1/4 cup water...
I'm thinking about calling it over for my 2017 varieties, at least for the test cakes I started. I want the 1/2 pint jars for this other stuff that's actually working. I'll let the plates I have sit as long as they want
Edited by Sidestreet, 29 April 2021 - 04:34 AM.
Posted 02 May 2021 - 04:08 PM
My PE test jars are showing growth:
So I did a little run of them to get cloning stock and maybe swabs:
And my Redboy MS jars are doing their thing:
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