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Cloning from dried P. Cyanescens


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#1 Traderkain

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Posted 18 July 2019 - 06:28 PM

So please let me be clear, I am in no way an experienced lab technician or clinically trained scientist. I am a new to all of this, but thankful for the info saved for someone like me to come along and use the knowledge that was already collected both the failures and successes. So I am open to advice for any of that view this and are willing to give advise. I see that we have a lot of people that view my threads but none post, I'm hoping you'll change your mind reading this. I was given specimens of "Oregon Blues" p.cyanescens this is what he called them. I'm still looking for the exact strain but without doing it underneath the microscope, I won't be able to ascertain the exact one. That being said I refuse to pay for Spores! I live in the state that has plenty of mushrooms in it so I did some searching around to try and find some but I received no feed back on what type of mushrooms they were so I threw them away out of fear of either an animal or child would eat them. I was blessed enough that when my friend found out what I was doing he gave me the mushrooms and told me to hydrate them for a print.. I told him "i'm going to attempt to cultivate these" with a little bit of a condescending laugh he said "good luck, my dad's best friend was a mushroom master and he could never get them to cultivate" with an even more condescending laugh I replied "challenge accepted". Now please before anybody comes in and starts tearing me down about the monumental task I've decided to undertake please note, I'm still going to try it. I felt I was fortunate enough for my friend to give me these and since I can't find anyone else on the Internet doing it I'm gonna make a thread about it. I've been thinking back to before my friend gave me the cyanes I can't believe how many places I stop and looked drive myself crazy trying to find something I can cultivate. I'm going to attempt to clone them in agar dishes using a couple different techniques I've learned on this site. I ordered them through Amazon the other night just waiting for them to get here. I'm so paranoid with using my credit card information over the Internet to buy things like this and have it shipped to my house. I made it an effort to learn everything from you guys on how to build my own substrate and will be attempting that as well in this thread. For me this is going to be a documentation of my successes and failures. Any and all help along the way would be greatly appreciated. I am hoping my successes are greater than my failures.
I will post photos from my phone in a few minutes wish me luck.

Edited by Traderkain, 19 July 2019 - 12:07 AM.

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#2 Traderkain

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Posted 18 July 2019 - 06:37 PM

Pictures of what I was given, well what's left of it :) I will post more pictures as things start to develop.

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Edited by Traderkain, 18 July 2019 - 06:40 PM.

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#3 ElPirana

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Posted 19 July 2019 - 12:18 AM

Not trying to dampen your enthusiasm in this experiment, but I don’t think you’ll have any success cloning. Maybe if you can get some viable spores, although I admit I’ve never considered trying to get spores from the dried fruits. I hope you get the results you desire!
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#4 Traderkain

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Posted 19 July 2019 - 12:46 AM

Thank you for the reply I was starting to think this is a ghost site with just lurkers...
I appreciate your apprehensiveness, and I realize the task im taking on is not an easy one.. but that is not to say that it can't be done or that it has not been done before. There are links all over the this site talking about cloning cubensis, so why not Cyanescens? I know the odds are stacked against me and I know I'm in way over my head, but you rest assured... Agar plates will be attempted to be cloned into, substrate will be attempting to be grown into. I realize all nature is acting against me, being that I have never done this before, I'm mostly assured failure. I can tell you this much, we live in an amazing era where we have library's at our fingertips. I plan on using every bit of the knowledge I have gained from this site in hopes to accomplish this monumental task. I hope you will continue to pop in from time to time and give me your honest feedback.

Edited by Traderkain, 19 July 2019 - 12:50 AM.

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#5 onediadem

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Posted 19 July 2019 - 03:01 AM

It is possible. You have to get agar and petri dishes. You will not be able to clone them as they way past that avenue, but you can retrieve spores. You will have to make up agar dishes, and sterilize, or buy some sterilized wooden q-tips. I would also minimize the nutes in the agar. You want to hydrate the spores and make the myc search for food. It will take much longer than a normal print depending on how old and dried out the spores are, You can soak a piece of mushroom and try to rehydrate it to try for a clone, but the odds are totally against you on that part of the adventure.

 

Either way, best of luck!

 

https://www.amazon.com/WEFOO-Microblading-Permanent-Supplies-Applicator/dp/B07RJSVMFW/ref=sr_1_34?keywords=art+pointed+qtips&qid=1563520868&s=gateway&sr=8-34

 

 

 

 

 

I use these for retrieving spores from dried mushrooms. The wooden stick makes them easy to pc without the worry of melting your sticks. I love these. Being pointed makes it easy to do so many different things in mycology wtth them. They are a very nice way ti store spores, and to gift someone a strain.


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#6 Traderkain

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Posted 19 July 2019 - 11:41 AM

Thank you for the reply here are the agar plates I purchased I hope they're the right ones...
https://www.amazon.c...=gateway&sr=8-3

Also thank you for the information the Q-tip method it was not something I had found on this site, but it will be added to the list of technique used on the cloning process. I do have a few questions with regards to this, both towards the storage and/or sharing strains. When you are using the Q-tip to collect, are you Using steril water on the Q-tip for collection or is it a dry collection process? Once you have collected the spores onto the Q-tip what do you do with them after that? Last one...once collected are you cutting off the cotton tip and storing them in collection vials like centrifuge sample tubes? Sorry for all the questions my mind is like a sponge right now and questions need answers..........
Thanks again for the reply

#7 hyphaenation

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Posted 19 July 2019 - 12:42 PM

Viable spores is the way to go there I agree. Easy to spread once at the mycelium stage. No advantage imo to taking a clone besides which it will take a huge amount of messing around likely leading to frustrating failure. Still though , good on you if you go for the clone experiment anyways.


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#8 hyphaenation

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Posted 19 July 2019 - 01:06 PM

Now having said that , a bunch of threads come to mind where people discuss this very issue and experiment. Some you may have seen before...there's gold in them there threads !
 

Cloning From Dried Tissue

https://mycotopia.ne...m-dried-tissue/

 

EQ pins from dry tissue Karo Tek

https://mycotopia.ne...html?1106008656

 

 

 

Posted on Tuesday, May 18, 2004 - 04:50 am: tree_s.png copy.png delmsg.png printer.png movemsg.png Yes, you can clone a dried mushroom just fine. I've even cloned ones that have been dried with dessicant and stored in a bag with silica gel for over a year. 

No special tek. Just mix up your agar a bit wet. Use 75-80% of the recommended amount of MEA in water. 

Watch for the myc to start growing. It usually takes a week or two to take off. The contams will be right behind, or sometimes even ahead of the mushroom mycelia. You have to watch very carefully for the first signs of mushroom myc, and transfer it to a new, waiting petri dish. Do this two or three times, and you'll be able to outrun the contaminants and get a clean culture.

 

 

Posted 02 February 2006 - 09:25 PM

By the way, if you want to make your dry tissue cloning life more simple; hydrate the tissue and apply enough to a dish to surfice. After you've applied the hydrated tissue to agar, cover the tissue with another layer of hot agar immediately. It eventually grows right through to the top layer cleanly and can be transfered to a new dish of agar for further use....

Hot agar technique provided from Stephen L. Peele of FMRC... 

Edited by hyphaenation, 19 July 2019 - 01:18 PM.

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#9 Traderkain

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Posted 20 July 2019 - 02:07 AM

Thank you for the reply and the information, you're right some of it I have already seen but the other one was new to me thank you for that additional link. ..
So here's what I did..
1. I made sterile syringes and let cool.
2. Made a Hillbilly glove box, out of sterile tote and cling wrap and sterilized with lysol...
3. Used cool sterile water to rehydrate mushroom caps (4) in foil.
4. Collected sterile water that rehydrating caps did not absorb along with spores suspended in the liquid.
5. Dry sifted caps for spores on to paper.

After I got everything sterilized and into the glove box I started with the cutting of the rehydrated mushroom caps putting small wedges into agar plates. (4)

Next agar plates with dried spores (2)

Next I cut dry mushroom cap wedges and sterile water put on top to hydrate while in agar plate. (2)

Last I used sterile liquid from first hydrating caps and sprayed on agar plates as LC.

I'm not sure how well I did on the process of sterilization but we will find out within the next couple days. Right now all cultures are done photos are attached thanks for all the advice now we will just have to wait and see

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#10 Foster

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Posted 21 July 2019 - 03:16 AM

Wish you much luck and success.  I once put a dried stone i a small amount of water, It was over a yr old and too tough to chew. I forgot it and a few weeks later when I remembered, it had myc growing. Also rehydrated a small chunk of wood with myc from Ps. Arcanus?  It grew myc fluff but I was unable to successfully transfer it at the time. Just about to transfer a few ropey myc strands of ps cyan to a few mediums myself. From a 5 yr old tray of almost totally decomposed wood chips .

 

I think the spores and cap pieces should have a chance !  I hope so.  Ive read about using activated charcoal in your agar in a few threads, to aid in germination of old spores.

 

My most successful transfers of this variety from agar was to Japanese knotweeed and PBH (par boiled rice hulls). It colonized the PBH (not sterilized/ pc'd) quite nicely. The knotweed as well.  https://mycotopia.ne...land-2012-2014/

 

Once it was on wood, I saw no contaminants.. none.. . Its been sitting in an open tray for a few yrs, in a dirty wet basement and still shows a little life.

 btw, PBH is supposed to be steamed sterile from the bag.

 

keep posting updates :)   and  Hit me with a PM ;)


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#11 Traderkain

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Posted 21 July 2019 - 11:25 AM

Can someone from admin please delete the empty picture post that were doubled I don't know why my phone keeps doing that. But here is an update first set of pictures are from 24 hours

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#12 Traderkain

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Posted 21 July 2019 - 11:30 AM

36 hours

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#13 Myc

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Posted 21 July 2019 - 11:49 AM

I'm not noticing any fungal mycelial growth in any of the above photos.

In the last set of images, that appears to be yeast (or possibly bacteria) growing.

 

Keep on trying. It's easier than it seems to clone from dried tissue.

 

Try trimming your tape into narrower strips so as to eliminate the small pockets from overlapping that wider tape.

Also, consider the expense of some bona-fide Parafilm. You won't regret the purchase if you intend to do much culture work. It's useful for tons of other applications as well.


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#14 ElPirana

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Posted 21 July 2019 - 12:58 PM

I really didn’t realize it was possible to clone from a dried fruit. I’m about to start growing again, but have to re-buy all my supplies. I thought I would not be able to grow again (for various reasons) and so got rid of almost everything last year, including gifting away all my cube spore prints. Now I’m down to the end of my stash.

I’ve always loved agar work so this sounds like a fun project to try. Just need to get enough supplies to go again.

@Traderkain I look forward to seeing your progress!
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#15 Alder Logs

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Posted 21 July 2019 - 04:32 PM

I just keep thinking, if you are on the Pacific Slope in the PNW, and you want P. cyanescens, just find a source of fresh wood chips, like where the power utility has chipped branches along a roadside.  Take chips and put then in shady grassy areas here and there and let Nature take its course.   I found my first P. cyanescens about thirty years ago, next to my chopping block where I split my alder firewood.  Later I found them where an industrial sized mower was regularly run under alder trees and the blades of the mower had repeatedly chopped some fallen limbs up.   Paul Stamets estimates that one in every four loads of wood chips delivered to someone's yard will produce P. cyanescens in Western Washington.  So, I say, "chip it and they will come."


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#16 pastyoureyes

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Posted 21 July 2019 - 05:02 PM

You could use saran wrap arount the plates instead of tape. Cut an end off the roll about an inch long with a saw.

I agree the 4th pic is likely bacteria/yeast.
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#17 coorsmikey

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Posted 21 July 2019 - 05:21 PM

If all the above fails. Simply take a clean pair of tweezers and break off a piece of gill material, the smaller the better. Then stab slightly into the agar. Do 6-10 plates this way and I will buy you lunch if you don't get at least one or two plates with mycelium to transfer clean. The may not be completely clean but your contamination will originate from a single spot as well as good mycelium. This method you may get spore germination or actual cloning from the tissue. When you add water to the dried mushroom to rehydrate you are just creating a substrate for the nasties to breed on.


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#18 Traderkain

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Posted 21 July 2019 - 10:53 PM

Thank you all for the post feedback and advise...

@Foster, thanks for this info I on the PBH I was going to attempt whole barley, rye berry and BRF substrate once I find my myc growth.

@MYC, thank you for you eyes, and knowing what to look for..I woke up this morning thinking I was going down the right path, after reading your reply I realized I had my first failed attempt, where this might deter a normal individual but it only strengthens my want for success.

@Elpirana I am still trying, I know it can be done, maybe with more skilled hands but I am determined to reach my goals. I hope all works out for you with your next run.

@Alder Logs, I will find them this fall as well as a few others I'm going to search for thanks for the advise, I'm going to get some wood from the miles gravel up here tomorrow and put a bunch in my back yard area that is still wild but secluded and see what chip it and they will come looks like...

@pastyoureyes, thank you for the glad wrap suggestion it would have been a better idea then using packing tape (it was all I had on hand), I will use that little tidbit if I cannot find parafilm tape.

@coorsmikey I will try the next order of plates like this and hopes that something will come of it. Thank you for the idea and the offer of lunch, btw I'm a buffet type of person FYI.
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#19 Alder Logs

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Posted 22 July 2019 - 12:12 AM

If you haven't got it, get Stamets' book, Mycelium Running. It has lots on easy ways to propagate any of the caramel cap wood lovers.

 

But that said, it just keeps blowing my mind how often they find the wood chips in a garden or flower bed.  Just last year, they came up strong in cardboard mulch under a neighbor's pear tree.

 

https://mycotopia.ne...ear-tree/page-1


Edited by Alder Logs, 22 July 2019 - 12:19 AM.

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#20 Traderkain

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Posted 22 July 2019 - 12:36 AM

If you haven't got it, get Stamets' book, Mycelium Running. It has lots on easy ways to propagate any of the caramel cap wood lovers.
 
But that said, it just keeps blowing my mind how often they find the wood chips in a garden or flower bed.  Just last year, they came up strong in cardboard mulch under a neighbor's pear tree.
 
https://mycotopia.ne...ear-tree/page-1


Thanks for the name, I came across another of his books online for free...felt I should share with this amazing site.

http://library.unite...oms_at_Home.pdf
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