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Anonymous
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 Re: Crossing two strains IS possible..Heres how...  [Re: i am e goldstein]
#1155696 - 12/19/02 03:52 PM (22 years, 1 month ago) |
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When you have mycelium isolated on agar, everything becomes apparent.
If you know you have a limited number of spores within a solution. You know the spores are not clumping, but are individual in the solution. (microscope needed)
You can place drops of this solution evenly scattered around a petri plate of media. Individual spores will germinate, and can be isolated from eachother before compatible monokaryons mate. You do this with each STRAIN you want to cross.
Once you have monokaryons of each, you can plate them up together(one from each strain next to each other).
Lots of them may just continue to grow two distinct SLOW GROWING monokaryons. Others will mate. Those that mate will have a single uniform dikaryon growing from the two matched monokaryons. This mycelium is faster growing then the monokaryons. You can assume this is the Dikaryon, or can make sure, by plating each suspected dikaryon up seperately.
Clamp connections are present in cubensis Dikaryons, so you can KNOW for sure that a Dikaryon has occured. USUALLY you can just TELL one has occured, because Dikaryotic mycelium LOOKS DIFFERENT then monokaryon mycelium. When everything is occuring in isolation(Dilution versus concentration) it all is Visibly occuring.
Each Dikaryon formed is tested for fruiting and spores. These spores formed are hybrid spores between TWO DIFFERENT STRAINS.
According to the literature, HYBRIDS tend to be less vigorous then each of the DONOR Strains. SOME are more vigourous.
Lots of work, and in the end you just have another CUBENSIS. According to the literature, Better off spending your time isolating the most vigourous substrain of your favorite Strains on/in your particular environment.
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 livingsublime
Stranger
Registered: 10/26/02
Posts: 555
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Re: Crossing two strains IS possible..Heres how... [Re: ]
#1156004 - 12/19/02 05:47 PM (22 years, 1 month ago) |
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Man this was a cool thread!
I just got done reading it all and it took a while but it was realy neat to hear all about what could posibly hapen or maybe not.
Teonan:man your smart,you have so much info it's not even funny!
How about you try and combine 2 different strain or whatever and post you results in the grow logs.
I would but I am a newbie to growing and still useing myco-bags!
Great read though guys!
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 em_bre_O
shroomery'sEmbryoticAsshole
Registered: 05/27/02
Posts: 2,312
Loc: In the stages of develope...
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Post deleted by Administrator [Re: ]
#1156009 - 12/19/02 05:49 PM (22 years, 1 month ago) |
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Skikid16
fungus fan
Registered: 06/27/02
Posts: 5,666
Loc: In the middle of the nort...
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Re: Crossing two strains IS possible..Heres how... [Re: em_bre_O]
#1156618 - 12/19/02 10:16 PM (22 years, 1 month ago) |
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Quote:
I am not trying to offend with this post but I personally need more proof than what is given here such as the ppl that have status outside of this tiny end of the net Meaning preferably Stamets.
See what Stamets has to say, read ch. 11 in TMC, especially the section "Spore Dilution Technique" on p. 340. Good Stuff. 
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Re-Defeat Bush in '04
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mycofile
Pooh-Bah
Registered: 01/18/99
Posts: 2,336
Loc: Uranus
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Re: Crossing two strains IS possible..Heres how... [Re: ]
#1157325 - 12/20/02 06:13 AM (22 years, 1 month ago) |
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Teonan, as usual, I completely agree with nearly everything you've said. Especially the criticism of PF's theory, I never liked it myself. I think it's funny how he uses such poor "scientific" tests for his theories, then he very condescendingly refers to people like us as "pop mycologists".
Anyway, a tip on spore dilutions. Run them through an eberbach container, or a blender jar ala 9er tek. The blending will break up spore clumps, but not damage the spores. Blending at each dilution stage helps alot. You will much fewer clumps which lead to the dikaryons if you blend the spores. For proof as to how effective this is at breaking up the spores, nanook is getting up to 100 spyringes per print with blended spore water, vs 10 or so with clumpy spore water.
I do wonder however about how common anastomosis is. I think it may be much more common than people think. But so far as usefulness goes, there is no good way aside from dna profiling to be certain when the genetic material has actually been exchanged. At least not that I know of. So, I think in nature and in practice the "non-self unions" may be rather common, but they are pretty useless when it comes to breeding.
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"From a certain point of view"
-Jedi Master Obi Wan Kenobi
PM me with any cultivation questions.
I just looked at my profile and realized I had a website at one point in time on geocities, it's not there anymore and I have no idea what I had on it. Anybody remember my website from several years aga? PM if so please.
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em_bre_O
shroomery'sEmbryoticAsshole
Registered: 05/27/02
Posts: 2,312
Loc: In the stages of develope...
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Post deleted by Administrator [Re: Skikid16]
#1157613 - 12/20/02 08:31 AM (22 years, 1 month ago) |
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Anonymous
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Re: Crossing two strains IS possible..Heres how... [Re: ]
#1158178 - 12/20/02 01:03 PM (22 years, 1 month ago) |
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- Post History Deleted Upon User's Request -
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i am e goldstein
Your Friend
Registered: 12/06/02
Posts: 283
Loc: 4 Blocks from Ground Zero
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 Post deleted by Administrator [Re: ]
#1158225 - 12/20/02 01:25 PM (22 years, 1 month ago) |
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Who controls the past controls the future. Who controls the present controls the past.
3DSHROOMS is either clueless or the worlds biggest SCUMBAG
I lie and exagerate constantly... God Bless America!
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Anonymous
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Re: Crossing two strains IS possible..Heres how... [Re: i am e goldstein]
#1158244 - 12/20/02 01:33 PM (22 years, 1 month ago) |
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- Post History Deleted Upon User's Request -
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Suntzu
Geek
Registered: 10/14/99
Posts: 1,396
Last seen: 25 days, 18 hours
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Re: Crossing two strains IS possible..Heres how... [Re: ]
#1158324 - 12/20/02 02:04 PM (22 years, 1 month ago) |
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Teonan, great posts, very informative;
I really like the suggestion of lightly blending the spore solution. One other thing that might make this acutally *possible* for a home cultivator with a little equipment. . . After establishing a collection of monokaryons, to use flat-bottom 96 well plates for the crossings; They would be easy to keep track of [A1-H12], and the flat bottoms are designed with some kind of light analysis [e.g. microscopy] in mind. A few drops of agar in each well [they come sterile] and you have a micro-petri. It would be cheaper and more compact than petris, and you wouldn't have the problem of one crossing accidentally 'mixing' with its neighbor. 96 crossings on one petri-sized unit 
Maybe they do this already? If I were to do it, that would be the route I'd go; With spore solutions at a proper blended dilution, a few sleeves of petri plates could generate enough monokaryons for a few 96 well plates; That's quite a few crossings, not a whole lot of commited resources;
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Anonymous
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Re: Crossing two strains IS possible..Heres how... [Re: Suntzu]
#1158338 - 12/20/02 02:13 PM (22 years, 1 month ago) |
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- Post History Deleted Upon User's Request -
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i am e goldstein
Your Friend
Registered: 12/06/02
Posts: 283
Loc: 4 Blocks from Ground Zero
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Post deleted by Administrator [Re: ]
#1158402 - 12/20/02 02:56 PM (22 years, 1 month ago) |
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Who controls the past controls the future. Who controls the present controls the past.
3DSHROOMS is either clueless or the worlds biggest SCUMBAG
I lie and exagerate constantly... God Bless America!
Edited by i am e goldstein (12/20/02 02:58 PM)
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Anonymous
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Re: Crossing two strains IS possible..Heres how... [Re: mycofile]
#1159069 - 12/20/02 07:44 PM (22 years, 1 month ago) |
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The addition of a surfactant to the water(soap,tween) has also been mentioned as a means to get non clumping solutions. But I suggest using them fast, soap seems to accelerate germination within the solution.
NON SELF UNIONS are the only succesful ones in nature. They are the ones that produce viable spores. No two monokaryons that are identical for mating type factors can mate and form a dikaryon.
Every Fertile dikaryon is the result of a non-self union.
Essentially, I wouldn't buy any HYBRID spores originating from any mushrooms that werent the result of MATED MONOKARYONS, of Different strains of KNOWN TYPE.
I would buy a culture of a PURE STRAIN of KNOWN VIGOR no matter how the Dikaryon was formed. Mated monokaryons or Anastamosis of Dikaryons.
I think when you multispore a jar of substrate, all of this occurs quite frequently. I think more then a single fertile substrain(Dikaryon)emerges from the chaos to yield the fruits. I also think that each fertile dikaryon probably has large sections of it's mycelial network made up of fused infertile(false matings), these sections just don't fruit. They can still be involved in digestion and transport(increased surface area of feeding). Many false matings and monokaryons also get over-run, and die off, because they could not fuse with the rest of the mycelium due to incompatability( self-unions). In the end a multispored jar of substrate is a collage of substrains fertile and infertile, all working together to make mushrooms.
I agree Anastamosis is probably extremely common when considering the number of spores that are involved in reproduction. It is kinda a catch 22, concentration increases the chances of success, but dilution guarentees KNOWING of the success.
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Anonymous
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Re: Crossing two strains IS possible..Heres how... [Re: em_bre_O]
#1159108 - 12/20/02 08:06 PM (22 years, 1 month ago) |
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How much will you pay me for STEP by STEP photos?
Stametes is the one who said the hybrids formed via either method were less vigorous then their donor strains MOST OF THE TIME.
The literature states that P. cubensis has a certain type of Breeding strategy, heterothallic, tetrapolar, SO SOMEONE MUST HAVE DONE A COMPATABILITY STUDY to KNOW THAT!!!
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Anno
Experimenter
Registered: 06/17/99
Posts: 24,168
Loc: my room
Last seen: 12 days, 16 hours
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Re: Crossing two strains IS possible..Heres how... [Re: ]
#1159335 - 12/21/02 01:10 AM (22 years, 1 month ago) |
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>How much will you pay me for STEP by STEP photos?
Is a sixpac enough?
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whiterasta
Day careobserver
Registered: 04/09/02
Posts: 1,780
Loc: Oregon
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 Re: Crossing two strains IS possible..Heres how... [Re: Anno]
#1159398 - 12/21/02 02:34 AM (22 years, 1 month ago) |
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Hell Anno make it a case! Great stuff Teonan especially outlining the types of mycelial connections.I learned some NEW info that explains some things I have observed.Thank You for the time to post it WR
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To old for this place
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Anonymous
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Re: Crossing two strains IS possible..Heres how... [Re: i am e goldstein]
#1159460 - 12/21/02 03:33 AM (22 years, 1 month ago) |
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- Post History Deleted Upon User's Request -
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ralphster44
collector
Registered: 01/03/01
Posts: 4,657
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Re: Crossing two strains IS possible..Heres how... [Re: mycofile]
#1159461 - 12/21/02 03:33 AM (22 years, 1 month ago) |
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Post deleted by administrator.
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WE SHIP TO CANADA FROM WITHIN CANADA
For your safety and security, we have a Secure Website.
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Your security and safety is of utmost importance to us.
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i am e goldstein
Your Friend
Registered: 12/06/02
Posts: 283
Loc: 4 Blocks from Ground Zero
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Post deleted by Administrator [Re: ]
#1159518 - 12/21/02 04:22 AM (22 years, 1 month ago) |
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Who controls the past controls the future. Who controls the present controls the past.
3DSHROOMS is either clueless or the worlds biggest SCUMBAG
I lie and exagerate constantly... God Bless America!
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Anno
Experimenter
Registered: 06/17/99
Posts: 24,168
Loc: my room
Last seen: 12 days, 16 hours
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Re: Crossing two strains IS possible..Heres how... [Re: i am e goldstein]
#1159573 - 12/21/02 05:04 AM (22 years, 1 month ago) |
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