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jimmyhabbitts
Stranger
Registered: 11/21/13
Posts: 3
Last seen: 9 years, 11 months
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Dikaryotic Mycelium
#19170097 - 11/21/13 01:22 PM (10 years, 2 months ago) |
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So Iam new to this site and apologize in advance if I am posting this in error in any way. I have a very simple question. In order to produce dikaryotic mycelium, it is said the petri of PDYA should be inoculated in four locations close to thecenter. The mycelium should grow radially and where the hyphae meet, somatogamy will occur and diakaryotic mycelium will result, correct? Is dikaryotic mycelium visibly different from monokaryotic mycelium? Also, do I wait for these initial petri dishes to become fully covered with mycelium if there are no other signs of contamination, or do I want to take samples of the mated mycelium and inoculate fresh petris? i plan on developing my own culture slants. Will these culture slants and spore syringes gotten from online vendors keep in a well sterilized refridgerator, and at what temperature? I am working in woodstove conditions in the middle of winter, so any advice towards that angle would be helpful too, though I ve taken adequate insulation measures, constructed and sterilized an jnnoculation hood, have all the proper equipment, inclhding fruiting chamber shelves, and everything going in or out of the lab is steralized at 15 psi kn a pressure cooker for one hour, etc. Two sonic humidifiers have been employed in conjunction with a fan which has been sterilized and affixed to the ceiling. Thanks in advance, and again, my apologies if I've made any newbie blunders.
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blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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ill help with what I know, which is not a lot, and hopefully a TC will chime in and offer some more enlightened guidance. As far as I know, monokaryotic mycelium would be hard to produce without a lab and microscope. monokaryotic myc is from a SINGLE spore, so if u are if u are inoculating with a print or syringe, its going to end up being dikaryotic, which is what you want if ur intention is to grow... (from TMC pg.27) "As a rule, monokaryons are not capable of producing fertile spore-bearing mushrooms. When two compatible monokaryons encounter one another and mate, cytoplasmic and genetic material is exchanged. The resultant mycelium is a dikaryon that can produce fertile offspring in the form of mushrooms."
I think consensus is do not let ur petris go to edges of plate, as the edges are where con tams are most present/most likely to enter, so for the first plate that was inoculated with syringe/print take transfers at first sign of growth. then plates after that, you can let them grow out a bit to see where the sectoring is occurring, and transfer the healthiest looking/most aggressive rhizomorphic mycelium from the edges of the growth. Do it from several spots, and eventually,as you do this, you will end up with an isolate. Because you are starting from MS, this isolate may not have the qualities you are looking for in fruit production. Most people, when trying to isolate for specific qualities will start from a fresh tissue clone, taken from a cluster, that way they at least know that when they begin to isolate myc from that clone (a few transfers down), it will be capable of fruiting, but may not have other qualities they are looking for (potency, size, speed, etc)
long term storage I have no idea. I keep them wrapped in petris in my fridge, but they will dry out no doubt in time, slants are way to go, Frank has a slant TEK http://www.shroomery.org/forums/showflat.php/Number/18182209#18182209 also RR's "Let's Grow Mushrooms" videos are a great start for everything you are looking to do http://www.mushroomvideos.com/
as far as you being in "wood stove" condition in middle of winter, I assume you mean its dry, humidity low. I remember RR saying he also lives in a cabin where it is very cold, and keeps a humidifier running 24/7. Ambient humidity in my apt is around 40% and my mono tubs and SGFC do well, if they are made to spec.
I keep my syringes in fridge in bag, no problems yet.
Like I said, I'm still new to agar, only been working with it first a month or two now, but its great fun, and hopefully someone more experienced will be able to help you out further. Good luck!
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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jimmyhabbitts
Stranger
Registered: 11/21/13
Posts: 3
Last seen: 9 years, 11 months
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Thanks. I was under the impression that mycelium from mushrooms belonging to the basidiomycete, like p. cubensis, began with monokaryotic otic mycelia and that is why the inoculations are done in several places in each petri, so that when the hyphae meet somatogamy is induced and the resulting mycelium will be dikaryotic...is it necessary then to inoculate more than one point in the agar. I am using grain media for the incubation period and was wondering if any of the environmental conditions should be altered to promote mycelium growth in the mason jars. I'm assuming the conditions would remain the same in terms of no light, high humidity, and a temperature of about 75 F. Is there anything that should be altered when going from petris to mason jar incubation of mycelium? Has anyone tried stopping the top of mason jars wi inth filters to allow CO2 circulation while at the same time keeping out contaminant molds? If so, what type of filters is recommended? And back to the mycelium, once it sectors, if I take a sampling of the rhizomorphic growth and inoculate it in a fresh petri will the resulting mycelium be rhizomorphic as well or will it continue to sector? Another question I had, and I'll have to look through the other threads to see if there is any info on this, but I am lead to believe from what I have read so far, that if you take dikaryotic mycelium from two separate petris and induce somatogamy, that a sub strain would develop. Is this true, or is it necessary to mate the mycelium from two entirely different strains to obtain a hybrid? In other words, if I wish to develop a hybrid strain of mushrooms, would I see a lot of genetic diversity from mating two cultures begotten from the same spore syringe or from differing ones, and should I mate the mycelium while both are monokaryotic or after a dikaryotic culture has been established? I want as much info as I can, just for intellectual purposes obviously lol, I have no intention of breaking any laws.......
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JMcDoogle
A Serious Scholar


Registered: 07/07/09
Posts: 1,495
Loc: Nunavut
Last seen: 3 years, 9 months
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Quote:
blindingleaf said: ill help with what I know, .......... Good luck!
Great stuff, * * * * *.
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The ego is nothing other than the focus of conscious attention.
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magickspore



Registered: 12/11/12
Posts: 798
Loc: Center of the universe.
Last seen: 8 months, 4 days
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Um I'd appreciate one of the more experienced cultivators to check my response or provide links that prove or disprove my response.
1 Light is not a factor and is beneficial to mycelium at all stages. The no light is old. Temp correct. During incubation you want to keep CO2 at the same levels as the introduction of fresh air is a pinning trigger. I believe there are many people that use SFD with success. Those can be bought at a many of the vendors. Not sure if that answers the CO2 question.
2. Mycelium can change from rhizomorphic to tomentose based on nutrients in the agar. Tomentose is not necessarily bad.
3. To create a sub strain you have to take the spores of a fruit and mate them thru multiple generations before an established sub strain becomes apparent.
Again I would appreciate anyone with more experience to post links either proving or disproving my statements above.
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 8 days
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Quote:
magickspore said:
3. To create a sub strain you have to take the spores of a fruit and mate them thru multiple generations before an established sub strain becomes apparent.
easiest way is to take a print of sa for kick sakes AA+ and print a redboy right on top of that print, use the print and the white fruit with red spores is the one you wanna clone
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
Edited by cronicr (11/21/13 10:11 PM)
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
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Re: Dikaryotic Mycelium [Re: cronicr]
#19172882 - 11/21/13 10:20 PM (10 years, 2 months ago) |
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Here is a good link to answer some of your questions OP. Also as cronicr suggested you would want to do this with at least one fairly unique variety (two would be better) on a macroscopic level, otherwise it would be pretty pointless.
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JMcDoogle
A Serious Scholar


Registered: 07/07/09
Posts: 1,495
Loc: Nunavut
Last seen: 3 years, 9 months
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Quote:
magickspore said: Um I'd appreciate one of the more experienced cultivators to check my response or provide links that prove or disprove my response.
1 Light is not a factor and is beneficial to mycelium at all stages. The no light is old. Temp correct. During incubation you want to keep CO2 at the same levels as the introduction of fresh air is a pinning trigger. I believe there are many people that use SFD with success. Those can be bought at a many of the vendors. Not sure if that answers the CO2 question.
2. Mycelium can change from rhizomorphic to tomentose based on nutrients in the agar. Tomentose is not necessarily bad.
3. To create a sub strain you have to take the spores of a fruit and mate them thru multiple generations before an established sub strain becomes apparent.
Again I would appreciate anyone with more experience to post links either proving or disproving my statements above.
There are a lot of questions there, its pretty much a threadjack.
Make your own thread perhaps to prevent deviation from the op's original topic.
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The ego is nothing other than the focus of conscious attention.
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blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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Re: Dikaryotic Mycelium [Re: JMcDoogle]
#19173771 - 11/22/13 06:14 AM (10 years, 2 months ago) |
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Quote:
JMcDoogle said:
Quote:
blindingleaf said: ill help with what I know, .......... Good luck!
Great stuff, * * * * *.
what do the stars stand for? hope its not bad... if it is, I apologize in advance for any misinformation, just trying to help with what I know and have read.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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JMcDoogle
A Serious Scholar


Registered: 07/07/09
Posts: 1,495
Loc: Nunavut
Last seen: 3 years, 9 months
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Quote:
blindingleaf said:
Quote:
JMcDoogle said:
Quote:
blindingleaf said: ill help with what I know, .......... Good luck!
Great stuff, * * * * *.
what do the stars stand for? hope its not bad... if it is, I apologize in advance for any misinformation, just trying to help with what I know and have read.
Its in reference to the 5 Shrooms I gave you as your newest rating.
J
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The ego is nothing other than the focus of conscious attention.
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blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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Re: Dikaryotic Mycelium [Re: JMcDoogle]
#19173896 - 11/22/13 07:26 AM (10 years, 2 months ago) |
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oh sweet! I was thinking in my head ***** could mean something bad... now I feel bad.. thx dude!
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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magickspore



Registered: 12/11/12
Posts: 798
Loc: Center of the universe.
Last seen: 8 months, 4 days
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How the fuck was I jacking his thread when I just answered all his questions and nothing more. Fucking troll.
Don't answer that its a rhetorical question.
Edited by magickspore (11/22/13 01:48 PM)
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