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seylm
still breathing
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Combining Strains
#12869037 - 07/08/10 06:09 PM (13 years, 9 months ago) |
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Just out of curiosity... What would be the best way to combine two or more cubensis strains to grow mushrooms whose spores would contain genetic material from both strains? I was thinking inoculating a jar with spores from both strains, or maybe mixing colonized jars of both strains into the same substrate. Someday I want to combine every strain I can get my hands on into one, if that's even possible.
-------------------- Easiest and Stealthiest Way to Grow Psilocybin "you may be wearing an Armani suit, but it's just pyjamas" - Mooji
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ninja cat 09
A paranoid android
Registered: 10/11/09
Posts: 4,170
Loc: Mexico
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Re: Combining Strains [Re: seylm]
#12869116 - 07/08/10 06:24 PM (13 years, 9 months ago) |
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seylm
still breathing
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1 post and 14 days to go I'll check it out in two weeks though, thanks.
-------------------- Easiest and Stealthiest Way to Grow Psilocybin "you may be wearing an Armani suit, but it's just pyjamas" - Mooji
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ninja cat 09
A paranoid android
Registered: 10/11/09
Posts: 4,170
Loc: Mexico
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Re: Combining Strains [Re: seylm]
#12869242 - 07/08/10 06:49 PM (13 years, 9 months ago) |
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Quote:
x7x_x7x said: hi!
last year i make a series of experiments inoculating the same jar with different LC's. all jars give nice fruits, some of them with pink/pale purple spore prints. i grow one of these with poor results, but questions remains and i want to grow it again. i keep some spore prints, if anyone here wants to research with these i'll ship them for free.
anecdotal, a mix between koh samui and GT give a first flush with very few fruits, no more than 5 or 6, those become very very blue, almost black, when longitudinally cut. was a very potent flush. never more these fruits appear.
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RogerRabbit
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Re: Combining Strains [Re: seylm]
#12870452 - 07/08/10 11:02 PM (13 years, 9 months ago) |
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Quote:
seylm said: I was thinking inoculating a jar with spores from both strains
The first thing would be to read up on what a strain actually is. A strain isn't spores.
A strain is dikaryotic mycelium, formed when two compatible hyphae meet and exchange genetic information. RR
-------------------- Download Let's Grow Mushrooms 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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dokunai
Cactus, Cannabis, Cubensis
Registered: 01/31/10
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Two compatible monokaryotic hyphae could also come from any of thousands of different potential mating types, most of which can get freaky with each other. Not to make it sound complicated.
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shymanta
Mad Scientist
Registered: 01/27/05
Posts: 907
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Re: Combining Strains [Re: dokunai]
#12917841 - 07/18/10 11:44 PM (13 years, 8 months ago) |
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To cross something like PE with something like OI, make a dilute solution of sterile water and the smallest amount of spores you can manage. Say, 1oz of water. Make one for each "strain". Then add a very small amount of the spore water to another sterile oz of water to further dilute it. Draw some up into a syringe and inoculate some petri dishes. Do this with each "strain". This process is meant to isolate the spores from one another in hopes of getting monokaryotic mycelium. Once you know you have monokaryons of each "strain", transfer the mycelium of both to a single dish. Do this multiple times. See if they mate. Test the daikaryotic mycelium for fruit-ability.
I've never tried this, but I've read about it from several sources and it seems feasible. I don't know, maybe you'll end up with Penis India.
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fastfred
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Re: Combining Strains [Re: shymanta]
#12922969 - 07/20/10 05:59 AM (13 years, 8 months ago) |
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Naw, that's just foolishness.
Isolate a monokaryon, colonize a jar with it, then inject fairly diluted spores from the other strain.
Workman developed this method and it's the easier than any method I can come up with.
-FF
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bw86
Doesn't play well with others
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Re: Combining Strains [Re: seylm]
#12923132 - 07/20/10 07:17 AM (13 years, 8 months ago) |
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Quote:
seylm said: Someday I want to combine every strain I can get my hands on into one, if that's even possible.
even if possible you would be back to square one. it would look like a normal cube and would have 1000s of phenotypes
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seylm
still breathing
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Re: Combining Strains [Re: bw86]
#12926042 - 07/20/10 05:36 PM (13 years, 8 months ago) |
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I don't care if the result has no distinctive or consistent traits. It will make me happy.
-------------------- Easiest and Stealthiest Way to Grow Psilocybin "you may be wearing an Armani suit, but it's just pyjamas" - Mooji
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Creepy John
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Re: Combining Strains [Re: seylm]
#12928540 - 07/21/10 05:40 AM (13 years, 8 months ago) |
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Sometimes just doing it is its own reward, Fast Freds method sounds like the easiest. Ive been wondering about the same thing who knows what the genetic dice will roll could be good or bad. I was under the assumption that strains produced this way typically don't fruit as well as either of the originals, so in my mind I'd guess you'd need to clone the fruits that you do get. Correct me if Im wrong here guys. I haven't tried this yet I'm just hypothesizing.
-------------------- Your perception is how you define your reality
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bw86
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i mean, you guys realize this is brought up every single day anyone ever see a novice succeed? just look at the Similar Threads below.
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idunno
PinkWebBuffalo
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Re: Combining Strains [Re: shymanta]
#12929387 - 07/21/10 10:34 AM (13 years, 8 months ago) |
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I've never tried this, but I've read about it from several sources and it seems feasible. I don't know, maybe you'll end up with Penis India.
DO IT! You will get an INDIAN PENIS STRAIN THAT WOULD RAWK_ LMAO(if you get one to grow, i want you to put a turban on it and take a pick 4 me,4 an avatar.
-------------------- The death of one man is a tragedy. The death of millions is a statistic.. Josef Stalin
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idunno
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Quote:
RogerRabbit said:
Quote:
seylm said: I was thinking inoculating a jar with spores from both strains
The first thing would be to read up on what a strain actually is. A strain isn't spores.
A strain is dikaryotic mycelium, formed when two compatible hyphae meet and exchange genetic information. RR
OK this may sound stupid but,here it goes.
According to the definition of a strain,in theory or what not-a spore print could contains prolly an almost infinite(trillions+)possible combinations, too make up a strain/substrain correct? Since it takes (2) monokaryon mycelium to make a dikaryotic mycelium.And dependending on the Dna sequence selected in wich order,also has an outcome.So to get a known trait,You would have to breed a dirayotic mycelium,to another single dikaryotic mycelium. Right? or no. Just to get predictiable results?
-------------------- The death of one man is a tragedy. The death of millions is a statistic.. Josef Stalin
Edited by idunno (07/21/10 10:51 AM)
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Creepy John
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Re: Combining Strains [Re: idunno]
#12931244 - 07/21/10 04:41 PM (13 years, 8 months ago) |
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Quote:
idunno said:
OK this may sound stupid but,here it goes.
According to the definition of a strain,in theory or what not-a spore print could contains prolly an almost infinite(trillions+)possible combinations, too make up a strain/substrain correct? Since it takes (2) monokaryon mycelium to make a dikaryotic mycelium.And dependending on the Dna sequence selected in wich order,also has an outcome.So to get a known trait,You would have to breed a dirayotic mycelium,to another single dikaryotic mycelium. Right? or no. Just to get predictiable results?
I don't think that's quite right as monokaryotic mycelia from a spore is haploid and has to 'mate' to produce dikaryotic mycellium capable of producing fruit, one cell with 2 nuclei and all the required chromosomes. Dikaryotic mycellium can fuse and exchange genetic material (anastomosis) but only if they are compatible. The reason mushrooms of a certain lineage produce similar fruits is most likely due to selective breeding limiting the possible genetic combinations.
Edit: Did a little rooting around in the archives and dug this up from 6 years ago.
Quote:
RogerRabbit said: As an alternative to putting two types of spores in a jar, try inoculating two different isolated strains of one species onto the same agar wedge. Simply transfer two wedges of myc to the same plate. When the two grow together, you'll see a 'line' form between strains. Sometimes one strain will gobble up and 'eat' the other one. Sometimes, each holds its own turf and the line stays put between them, where each has its own territory on the petri dish. About one time in a hundred, along this 'line' a third sector will open up. This is by definition, a 'hybrid' produced by dikaryotic pairings from each strain. It is definately and absolutely possible, and I've done it. Next fall, when I have time to resume the hobby, I'll do it again and write a photo documented tek. Until then, anyone who has time and inclination to experiment, now knows how to go about it.
-------------------- Your perception is how you define your reality
Edited by Creepy John (07/21/10 06:24 PM)
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fastfred
Old Hand
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
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> try inoculating two different isolated strains of one species onto the same agar wedge.
You'll not often end up with crosses that way.
As I mentioned Workman came up with the brilliant solution of colonizing with a monokaryon and then adding spores. That's how he produced his PE x albino cross.
The idea is that a monokaryotic culture will not fruit. Since the monokaryon from one strain has already colonized the substrate, when you introduce the spores chances are that they will mate with the monokaryon long before they meet up with another spore from their same strain.
Quote:
According to the definition of a strain,in theory or what not-a spore print could contains prolly an almost infinite(trillions+)possible combinations, too make up a strain/substrain correct?
Lots of foolishness about what a strain is gets posted on the shroomery.
Designating something a strain is just a labeling convention for working with any sort of culture. There's no hard and fast definition on how exactly to go about applying that label. You just use what is convention, what is reasonable, and you use whatever your best knowledge of the culture is.
Everything comes from nature at some point. When you collect something from the wild it's a "field sample". When you culture it you assign it a species. You usually don't have any genetic information on the cultures so you refer to them in a way that makes the most sense. Samples from the same location are assumed to be the same or very similar so you call them the same strain. Samples from different locations are assumed to be somewhat different, so you assign them different strain names. If you can observe reliable traits specific to certain samples you call that another strain.
Strains are analogous to human races or ethnicity. It's the same thing for other organisms too. You could call someone of the white strain or of the black strain. That's easy enough to see and justify genetically. But you also make assumptions based on location, just like you have a Chinese strain and a Japanese strain. Maybe you can't tell them apart by what they look like, but you assume they are different because they come from different locations. BTW, asians can tell each other apart very easily. It's a good example of significant genetic differences being easily overlooked, which is why you assume different locations to be genetically different.
So cultures with identifiable phenotypic differences are different strains. Cultures collected from different locations are also assumed to be different strains, until/unless you develop reason to believe they are the same as another strain.
Random spores or spore matings are referred to as "individuals", their spores (children) are "progeny" (offspring). The term "substrain" is more vague and you would usually just use that to indicate the lineage of the culture.
-FF
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idunno
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Re: Combining Strains [Re: fastfred]
#12934619 - 07/22/10 09:34 AM (13 years, 8 months ago) |
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So, really most,"strains" are just pheno types. Like corn in iowa,They test corn in fields, for next years grow to see which phenotype,grows fastest,most resistant to bugs,yield etc. Its all thee same "strain"of corn, but seed to seed has slight different charasticts, That could be bread with another that is similar to produce a redundant trait,which gives it uniform morphological or physiological property,makes it a Strain. Is this correct,or am i way off?
-------------------- The death of one man is a tragedy. The death of millions is a statistic.. Josef Stalin
Edited by idunno (07/22/10 09:40 AM)
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fastfred
Old Hand
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Re: Combining Strains [Re: idunno]
#12935571 - 07/22/10 01:34 PM (13 years, 8 months ago) |
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Sure. With plants you would probably call it a "cultivar" or "variety". Strain applies just as well though.
-FF
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Creepy John
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Re: Combining Strains [Re: fastfred]
#12936603 - 07/22/10 05:07 PM (13 years, 8 months ago) |
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Quote:
fastfred said: > try inoculating two different isolated strains of one species onto the same agar wedge.
You'll not often end up with crosses that way.
As I mentioned Workman came up with the brilliant solution of colonizing with a monokaryon and then adding spores. That's how he produced his PE x albino cross.
The idea is that a monokaryotic culture will not fruit. Since the monokaryon from one strain has already colonized the substrate, when you introduce the spores chances are that they will mate with the monokaryon long before they meet up with another spore from their same strain.
-FF
I agree that Workmans method sounds like the best way to go as I posted higher up in the thread, I was just rummaging around when I found RR's quote and thought hey there's a way ppl without a scope could try. RR even states it's like a one in a hundred shot.
Fred you have some experience trying Workmans method? In your opinion do cultures crossed this way produce average yields, compared to their progenitors? Or is there a lot of clone-isolate grow repeat to get yields back up to par?
-------------------- Your perception is how you define your reality
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fastfred
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I have tried Workman's method. IME it's a total crapshoot. Depends a lot on what you're trying to cross. Without genetic markers like PE or albino traits it's also impossible to verify that it's a cross and not just more of the same. Even then you usually have to wait for the crosses progeny (F2) to see most (recessive and additive) traits.
For the most part a cube is a cube, so it's not easy to get any amazing results.
-FF
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RogerRabbit
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Re: Combining Strains [Re: fastfred]
#12938925 - 07/23/10 12:19 AM (13 years, 8 months ago) |
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When monokaryotic mycelium 'digests' (out of lack for a better term) non-germinated spores and becomes dikaryotic, you're still really back to square one. There's uncounted genetic possibilities, and what you get is a dice roll.
When two isolated strains of dikaryons are crossed, the result is far more predictable. It doesn't work out 100% of the time, but what does? Timing is everything. You must be right there ready to grab the cross within hours, or one or the other strain is likely to overrun it and it will be lost to the much more rapidly growing established mycelium.
I've succeeded lately in crossing several commercial shiitake strains this way, and am currently testing them. I'm looking to find or create a strain that will fruit happily on softwoods, since that's what predominates in my area. I can get as many truckloads of Douglas fir sawdust I want for free, so there's a very lucrative application for finding a good producing strain that will grow and fruit on it. Getting that lucky from spores would be like hitting a jackpot in 'Vegas ten times in a row. RR
-------------------- Download Let's Grow Mushrooms 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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idunno
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If a cube, is a cube, is a cube, then they would all be basically the same strain. Just different pheno types,and sub types. So IMO you are really "breeding" for a recessive trait. Ie, (2) dikaryons the posses,Bacteria resistance,or rapid growth etc. Or (phyicial)mutations, that occurs, like Albino, different spore colour etc. I think that in theory, A true new strain would be sterile"like a mule,horse donkey hybrid" but it should be able to be cloned.Or it could be like hybrid fruit varieties(plumcot)(plum+appricot)and grow. give it a whirl,any thing could happen.
-------------------- The death of one man is a tragedy. The death of millions is a statistic.. Josef Stalin
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fastfred
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Re: Combining Strains [Re: idunno]
#12945949 - 07/24/10 02:35 PM (13 years, 8 months ago) |
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> If a cube, is a cube, is a cube, then they would all be basically the same strain.
As I went into earlier, it is standard to take samples collected from different areas and name them as strains. Just because you don't see much difference doesn't mean that they are all that similar genetically.
The example I used earlier is that most white people would just lump Asians together as a race. However if you follow proper methods of considering them different based on location you get a much more correct grouping that represents the differences. A Chinaman can easily tell a Korean from a Jap just by looks. Just because the differences are not obvious to you doesn't mean they don't exist.
> A true new strain would be sterile "like a mule,horse donkey hybrid"
No. That is an interspecies cross. And they don't always result in sterile offspring like donkey + horse does.
-FF
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