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Next step after cardboard cloning?


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#1 wumpsdad

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Posted 03 April 2005 - 05:50 PM

I have read that spores can be started on grey cardboard, so can mushrooms be cloned on it? would the same nutrient recipe be used?

And while this newbie is asking questions; is it possible or practical to clone direct to say a grain substrate? Or is it necessary to clone in liquid or agar first before expanding spawn?

Where are flow hoods necessary? for what steps in cultivation? are they necessary for grain expansion to more grain or straw? Would a heavy shower of negative ions do the job equally well?
I put an ionizer in our kitchen and after a few moths it dawned on me that we never had food spoiling in the fridge as we had before. Surely this must be useful for mushroom cultivation.
Any comments?

Thanx,

Wumpsdad

#2 Lazlo

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Posted 03 April 2005 - 06:14 PM

Sure you can definately clone to straight grain using tissue. I did it in a 1/2 pint jar of wbs for faster colonization, then g2g'd to 2 1qt jars, and then on to 3 dozen more. Works pretty good if your clean. IMO Karo is great for cloning though. As far as flow hoods are concerned, they're fanned HEPA filtrated devices for a sterile work areas. I don't have any pics, but i'm sure some will pop up. Don't attempt any cloning without a flowhood or gloove box.

#3 Guest_Peter Cottontail_*

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Posted 03 April 2005 - 09:13 PM

It would be best to always run your cloned material through antibiotic agar before placing into grain. I've cloned a lot of mushroom fruit bodies before and I don't remember any that were clean on the first go.

When I pick wild mushrooms for cloning, I always take along some moist cardboard. This is much easier and more forgiving in the woods than agar. On a two or three day trip, it's not unusual for the fruits to begin sprouting mycelium into the cardboard. When I get home, some of this fresh mycelium is transferred to agar to give any contaminants present a chance to grow, so I can then transfer away from them. Were you to clone straight to grains or liquid, the contaminants are as likely to grow as the mushroom tissue. Good luck!
RR

#4 wumpsdad

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Posted 13 April 2005 - 08:03 AM

Well, following Rodger's suggestion, today I cut off some of the new growth and put it on new cardboard. I followed basically the same procedure except for - oops!! I put 2 samples onto cardboard I had wet but forgot to H2O2 treat first! so I retrieved them almost immediately and washed the new cardboard down with 3% H2O2, and rinsed it after a minute.
So these didn't have the same period of H2O2 on them, and are potentially contaminated. I cross my fingers and hope that the reputation of Oysters as being vigorous and aggressive colonizers lets me get away with it.
I placed the transfered bits of cardboard face down as I was thinking this would speed colonisation- is this correct? or are they better left face up in the air?


........Aside: It has occurred to me that since I lack petri dishes, these sterile bags are an adequate substitute, tho a little fiddly.

I took the plastic bag which contained the cultures that had and hadn't been H2O2 dipped, and as best I could scraped away the visible original mushroom tissue. This was to prevent rot/contamination.
It had turrned slushy. I guess this tissue, differentiated as it is as fruit tissue, is not up to staying healthy as spawning tissue.

I added new pieces of H2O2 treated cardboard to the original culutres in jars to give them something to grow onto now that I have cut big bits away.
- Was this the right thing to do? at worst I suppose it may contaminate, or just waste time if it doesn't colonise.

I also cut cardboard from the piece colonised by the piece of mushroom cap
I put one cut off piece directly on fresh coffee grounds/filter paper in a fresh plastic bag, and others on more cardboard in bags.
However, since these pieces grew simply from a slice of cap, whats the chance that they also contain culture of germinated spores and not pure culture? I shall keep everything seperate and isolated.

I put a dozen or so pieces of thick grey cardboard into 3% H2O2, and put them in a plastic bag for later. When I need to start a new culture, I'll just rinse them off with cool boiled water first.

Well thats all so far.

Do mycelia "run out of steam" or something? is it due to a medium not providing sufficient nutrients? This is happening to my wild Oyster culture what is a good all round nutrient to add? I was thinking of beer broth[ 1 part beer, 3 parts water, PCed] or dissolved multi or B vitamin. Can I spray that on the medium? or dunk it?
I have had a few brainwaves that I will keep to myself for now and try out later. If they work you will read it here first.

Please add your comments.

cheeers,

Wumpsdad

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#5 Hippie3

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Posted 14 April 2005 - 06:32 AM

since these pieces grew simply from a slice of cap, whats the chance that they also contain culture of germinated spores and not pure culture?


zero

#6 Hippie3

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Posted 14 April 2005 - 06:33 AM

Do mycelia "run out of steam" or something? is it due to a medium not providing sufficient nutrients? This is happening to my wild Oyster culture what is a good all round nutrient to add?


no beer unless you boil off the alky
i'd try a dilute malt extract
but may well increase contam risks

#7 wumpsdad

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Posted 14 April 2005 - 03:59 PM

Thankyou Hippie3. I don't wanna keep asking dumb newbie questions, but what dilution of Malt?, or direct me to such an info store at this or other site?

Thanks,

Wumpsdad

#8 wumpsdad

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Posted 05 June 2005 - 08:51 AM

HI all,

I am still a beginner but gaining experience slowly.
I have for the second time cloned store bought Oysters with a view to growing them.
So far they are only growing as mycelia on cardboard.

How long should the mycelia grow before cutting it away from the original clone sample and growing it on to another medium? The original samples were taken on 30 May, a monday and today its 6 days, a sunday, 5,june.

I used a different cardboard than the first time and it seems not to have grown so fast. I tried water on the cardboard, and MultiB vitamin solution, and a combination.

How should I grow out the samples? put on further cardboard with other nutrients?
I have taken some old muesli- perhaps I should use fresh?- and its a mix of wheat, oats, and barley seeds, rolled, and put it in boiling water and let it cool down . I was going to let it soak for 24 hours then should I PC it?
I also chopped up some straw from a nearby farm. I wont wet it till I am sure I am ready to begin spawning.

I am just not sure what steps to take and when, and I don't want to waste my efforts. The last time I did this the mycelia didn't survive the transfer to more cardboard. I suspect it was due to a lack of nutrient which is why I used MultiB vitamin solution this time.

any hints or directions would be graetfully received.

Thanks y'all,

Wumpsdad

#9 Guest_Peter Cottontail_*

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Posted 05 June 2005 - 10:22 AM

I use cardboard when collecting wild specimens in the mountains. The mycelium will usually begin to grow on it, but as you found out it has very little nutrients. This is good when taking wild specimens because molds also have a hard time growing on it. Transfer the first few bits of clean growth you see to agar. Have several petri dishes poured and ready, because as soon as your mycelium begins to grow, you need to make another transfer of the brand new leading edge of the mycelium. Do this before any contamination shows up in the agar. Once you have a pure, clean specimen on agar, transfer it to rye grain to grow out. Use the rye grain to inoculate more rye grain, then spawn that to your straw. You're off to a good start it would seem.
RR

#10 wumpsdad

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Posted 05 June 2005 - 05:37 PM

Thanks for that info Rodger.

I will make it a point to cut from the leading edge. I should cut the leading edge of the cardboard, yes? and then to agar, and then cut the leading edge to a new agar.
I think I noticed from my first attepts that the bigger the initial sample, the faster and further the mycelium grew. Must be coz it had a bigger reserve of nutrients.
I had attempted to transfer the growth directly to Coffee grounds with filter papers but it just didn't happen, and I didn't know why then but as mentioned, now suspect it was lack of nutrients.

I don't have petri dishes. I have jam jars. I can get Agar, Malt/extract and Karo from the local Asian supermarket.

Any particular tek or class of tek i should follow?

Would a liquid culture attempt be too optimistic at this stage?

hope you like the questions 8-) !
I am enjoying adapting and improvising...sure is a challenge

thanx again,

wumpsdad.

#11 Guest_Peter Cottontail_*

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Posted 05 June 2005 - 06:02 PM

You won't be able to use liquid culture on a wild specimen until it's been totally cleaned up with agar. You need the two dimensional space of a petri dish to isolate your mycelium away from contaminants. You'll find bacteria on wild specimens to be as bad or worse than molds. I wouldn't take any of the cardboard to agar. Simply flame sterilize a scalpel or knife, then cool it in the waiting petri dish. Just scrape a tiny piece of the very leading edge of the mycelium from the cardboard and place it on the agar. Give it 48 hours until you just begin to see growth then take the very leading edge of that new growth and transfer it to a new petri dish. What you're trying to do is get ahead of the contamination before it sporulates. Do a search for agar in the archives and read all you can. Good luck.
RR

#12 Hippie3

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Posted 16 October 2005 - 10:00 AM

copied to the vaults




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