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Invisibleshobimono
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Hericium cloning
    #6205522 - 10/24/06 09:53 AM (13 years, 8 months ago)

I tried some on cardboard and honey water and didn't have any luck.
Is it possible to clone these without agar?
What part should you use? I tried using both part of the main fruitbody and a part of butt that was attached to the wood, but neither took on cardboard.


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OfflineRogerRabbitM
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Re: Hericium cloning [Re: shobimono]
    #6205574 - 10/24/06 10:10 AM (13 years, 8 months ago)

You really need antibiotic agar. Hericium is one of the hardest fungi to clone that I've ever worked with. The problem is that bacteria are completely infused into the fruiting body and attached to individual mycelium cells, so that no matter where you cut the material from, you get lots of bacterial rods which then take off and grow, overwhelming the fungi.

The success I've had with this genus has come from placing on antibiotic agar, then when the bacteria begins to grow, usually even before the mycelium starts, pour piping hot agar over the top of the growth, covering the bacteria and the tissue sample. The hot agar 'pasteurizes' the bacteria, knocking it down for a few days, but doesn't seem to harm the hericium tissue. You'll have to repeat this process several times before you have a clean culture.

Nice to see ya around again shobi. How was morel hunting last spring?
RR


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Invisibleshobimono
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Re: Hericium cloning [Re: RogerRabbit]
    #6206468 - 10/24/06 02:05 PM (13 years, 8 months ago)

Bacteria infused throughout the mycelium makes sense and would explain the bubbles that started coming off the inner stem piece when placed into honey water.

What about starting from spores?

Morel hunting last spring sucked, didn't find a single fruit. The summer on the otherhand was awesome, we picked from the first week of June until the first week of August


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OfflineFreeSporePrints
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Re: Hericium cloning [Re: shobimono]
    #6206514 - 10/24/06 02:15 PM (13 years, 8 months ago)

hi Roger have you pics of this tip?

do you puour only hot water + agar or the entire hot media?

thank you


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OfflineRogerRabbitM
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Re: Hericium cloning [Re: FreeSporePrints]
    #6206983 - 10/24/06 03:59 PM (13 years, 8 months ago)

I mix up another batch of antibiotic agar using gentamycin sulphate, and pour it over the culture while still very hot.

I've filmed this entire process and a tek for it will be on my upcoming DVD.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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OfflineHotnuts
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Re: Hericium cloning [Re: RogerRabbit]
    #6210179 - 10/25/06 10:32 AM (13 years, 8 months ago)

Here's a Lion's Mane and Angle's Wings I did the same way Roger's explaining. It's much easier when you first clone the specimen. Apply a very thin layer of agar before you inoculate the plate, just enough agar so the specimen can grow out. Then as soon as you see mycelium from the mushroom, apply another layer of very hot agar over the whole works, covering the growth and first layer completely. Contaminates and all. The mycelium from the mushroom will crawl through to the top layer, leaving behind the contaminates. You'll need to transfer the new growth to a new plate. It doesn't hurt to do the exact same thing with the new transfer either. Give the transfer a couple of days to start growing and then cover it with hot agar again immediately. Just let the plate colonize completely for use.

The mushroom mycelium will bubble up from the top layer of hot agar that's applied, leaving you to think that you've zapped and killed the mushroom mycelium. Don't worry, you haven't. See the bubbles in picture 2?





Edited by Hotnuts (10/25/06 11:04 AM)


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OfflineFreeSporePrints
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Re: Hericium cloning [Re: Hotnuts]
    #6210455 - 10/25/06 12:33 PM (13 years, 8 months ago)

hi Hotnuts hi Roger, thank you for the explanations, unlucky i can't see anything (except the bubbles) from the second pic.

just a question: during the transfer how is possible to take just the mycelium without the "downfloor" contaminants?


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OfflineHotnuts
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Re: Hericium cloning [Re: FreeSporePrints]
    #6210467 - 10/25/06 12:37 PM (13 years, 8 months ago)

If you look below the bubbles, you'll see where the mushroom mycelium has crawled up to the new layer of agar. Just take a sterile scalpel or whatever and use it to simply take a swipe of wispy mushroom mycelium for transfering, not the agar it's growing on.


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OfflineRogerRabbitM
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Re: Hericium cloning [Re: Hotnuts]
    #6212401 - 10/25/06 10:01 PM (13 years, 8 months ago)

A good analogy would be 'shaving' just like you remove hair from your face without taking the skin off too.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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Invisibleshobimono
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Re: Hericium cloning [Re: RogerRabbit]
    #6265036 - 11/09/06 11:20 AM (13 years, 8 months ago)

I forgot about this bag. I placed like 5 or 6 pieces of hericium in between the cardboard when I made it.

Does this look like one of the pieces actually made it?




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OfflineRogerRabbitM
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Re: Hericium cloning [Re: shobimono]
    #6265410 - 11/09/06 12:58 PM (13 years, 8 months ago)

Possibly. I'd take a piece from each of those growths and transfer to agar to grow out. Hopefully, one of them is the hericium and not mold. Hericium is a tough one, normally from bacteria though, not molds. The cardboard may have selected away from the bacteria problem because I don't see any slime.

Hericium mycelium smells strongly like fresh mushrooms the way cubensis does, so you might take a whiff and see if one of the growths has that very memorable scent.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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Offlineflower
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Re: Hericium cloning [Re: RogerRabbit]
    #6286750 - 11/15/06 05:07 PM (13 years, 7 months ago)

One thing that seems to be working for me is to shave the clone specimen thinly and place it in a shot-glass of peroxide. Let it fizzle for several seconds and transfer to liquid culture. I had a couple of jars contaminate (but I didn't let it fizzle as long as I usually do) but a couple look really good. I took the clone from a young Hericium.


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Mushrooms, Mycology and Psychedelics >> Gourmet and Medicinal Mushrooms

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