invitro, agar, etc.

[chrisandstepheny]

I am pretty new to mushroom cultivation. So, I wanted to see what all the other cultivators here had to say about this jar.
I am following a detailed plan right now. This is just a side project to that.
What I am about to show you is a mycelium culture on a agar substitute in the bottom of a half pint jar. It is a test jar for a LC. The strain is Z from a syringe.
What happened is, I was following my plan and testing my LC(2 strains 2 jars each with 1cc of LC each) on an agar substitute(basically powdered long grain brown rice and water pressure cooked into a little cake.) Then a couple days after I inoculated I left town for a couple weeks. While I was away my husband noticed that the agars were not colonizing with anything. So, he reinoculated one jar each of the two strains. The Z took off and the other strain contaminated. BTW, the other 2 jars eventually colonized with myc.
What we have been doing is opening up the jar a few times a day, keeping the myc hydrated, and exposing it to light to see if it will pin. As far as I can tell the jar has looked about the same as in the pictures as it did about a week ago when I got home.
What is everyones thoughts? I think the myc looks really cool and it would be even cooler if I could have a sample of shrooms before my main project is producing.

[BuckarooBanzai]

It might pin, if it stays contam free. You won't need to hydrate/add fresh air. It takes a while, but I've agar Petri dishes sealed with parafilm pin.

But ya won't get too much!

[Cardboard]

I have a similar jar going, BRF cooked in a 1/2 pt jar. Transfered some B+ spores, grew for a week or two so untill cake was fully colonised. Dunked it just yesterday and now it is sitting in a jar (4 holes in lid) under lights. Hoping it will pin soon

If you are interested I can keep you updated or grab some pics :P

[chrisandstepheny]

hey Cardboard, sounds like a neat little in vitro project you got. I am always ready to observe and listen to other's experiences. I am especially intrigued by in vitro. Kind of why I am still messing around with this jar.

BuckarooBanzai, I was reading up on some of the agar threads and seen some agars that pinned all on there own. I think if I don't decide to try a tissue culture with the other agars I will leave them alone and just observe their progress.

[crazy1]

I know a good strain will fruit on agar, have done it many times.
But each time it was a plate that contained an isolate of a strain I was working with at the time.
So it was the strongest rhizomorphic myc that fruited.
But as BB stated there's no need to keep opening up the jar, IMO all you're doing is increasing your chances for contams.
Good Luck to you 2

[limeade]

the only few things i can say is good luck, and i want some of that zstrain isolate, i can't believe it's made it this far without contamination.

[Cardboard]

Originally Posted by Cardboard

I have a similar jar going, BRF cooked in a 1/2 pt jar. Transfered some B+ spores, grew for a week or two so untill cake was fully colonised. Dunked it just yesterday and now it is sitting in a jar (4 holes in lid) under lights. Hoping it will pin soon

If you are interested I can keep you updated or grab some pics :P

The cake has pinned!! Found 4 nice looking fruits pokin out of the cake pictures will come when they grow larger

[chrisandstepheny]

Originally Posted by Cardboard

The cake has pinned!! Found 4 nice looking fruits pokin out of the cake pictures will come when they grow larger

Sweet, can't wait

[crazy1]

just found the pic of some fruiting on agar.
Here ya go.

[eatyualive]

cool pic crazy1.

nice pics cns...

[Cardboard]

Bit difficult to see the pins through the glass, thought it might be helpful to post a pic of the setup. I attempted to use a pop bottle lid as a riser but the cake was a little tight, ended up putting a hole in it (i think its really helping with ventilation). The cake is wedged up above the lid about 1/4''. A few more pins have sprouted, looks like 5-6 fruits now. After the first flush, should I expect a 2nd? Could I break the cake apart and case it?

[Freaky]

Fruiting out on agar is a good indication to help show you how pinning/fruiting will occur. I found it best to make a transfer so I know my mycelium is nice and healthy and clean, then fruit out the agar. This way I know which isolate/clone agar dish to inoculate with.
Examples below.



from this thread:
http://forums.mycotopia.net/agar-str...r-pinning.html (Agar Pinning)

[Cardboard]

Will most agar plates fruit if left for long enough in proper conditions, will most of these fruits be aborts?

[Freaky]

I'd say 90-95% of my agar dishes have pinned if I've left them alone.

It's a helpful way for me to test the fruiting/pinning type of isolate or multispore mycelium I'm working with.

I don't think there can be 100% guarantee put on the fact that the agar dish is going to pin though.

[chrisandstepheny]

Hey crazy1, I just thought that pic was pretty sweet. I had to nominate for pic of the month. I hope you don't mind.

[Freaky]

Agreed! That's a bad ass pic Crazy1! I'm glad to see you back around

[crazy1]

Thanx for all the props

It's great to be back Freaky

I have a lot of plates pin, but it's also a sign of a good strain to let run IMO
But as stated before most of the plates are 3rd or 4th transfer of rhizo myc

[Cardboard]

How far do these pins develop?

[Skatterman420]

thats really cool

[chrisandstepheny]

This is the other Z strain agar from the same LC as the agar I started the thread with. I think it might be contaminated which worries me since I already inoculated some jars with this LC. The agar I am about to show you was inoculated on July 13 and didn't show any growth until about August 8 when I noticed a small ball of myc in the middle where I dropped the lc and at the edge (where it is colonized up the glass) because I accidentally tipped the jar slightly after inoculating.
It kind of just looks like a shadow in the pic and it's not really much darker in person. There is little bits of powdered grain stuck to the inside sides of the jar making it hard to see. I have never opened this jar since sterilization.
To me it looks like a dark blueish or grayish discoloration of the myc. If you couldn't tell it is right in the middle of the bottom of the jar.
Also, I added a pic of the bottom of the jar. I thought that is might have grown a contam underneath the myc which happened to us before. I only see myc growing through what I guess are pockets of air cause I don't think the myc can grow through this stuff. But where you see the white spots on the bottom is directly under the dark spot.
I have a hard time believing this LC is contaminated, but what is going on?
If it is just bruising how did it get bruised?

[crazy1]

hahaha you guys rock going for the full blown agar grows

Now if it is bluish gray like this



your all good to grow


And the small specks you may see through the agar are undissolved particles IME

[chrisandstepheny]

You know I think it is just bruising. It freaked me out though. It did get hot in our incubation cabinet. I think I stalled my jars cause of that.

[goldenteacher1163]

chrisandstepheny IMO it might take a little while for the pinning to start because the myc is going to want to fully run through the agar substitute because it is not as solid as an agar plate would be making it more like a cake but this might also allow for more pins to mature
good luck

[chrisandstepheny]

Sorry, no pic, but the second "agar" that is in the pt jar did produce an abort. My husband ate it, and he said he did feel a little different from eating it.