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OfflineLochRaven
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dikaryotic mycelium questions
    #5533960 - 04/19/06 03:14 PM (17 years, 10 months ago)

when large amounts of mycelium grow is it only from the fusion of two monokaryotic cells? do multiple dikaryotic cells join? or does only one dikaryotic structure grow into the entire network of mycelium?

sorry if this doesn't qualify as advance mycology. i just didn't want to waste any time in other forums...


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Re: dikaryotic mycelium questions [Re: LochRaven]
    #5534214 - 04/19/06 04:30 PM (17 years, 10 months ago)

spores germinate into monokaryons. Two compatable monokaryons will mate to form dikaryons.
monokaryons can mate with dikaryons if compatable.
Dikaryons can mate with dikaryons also, anastamosis.

A colony can be made up of a single dikaryon, or many seperate ones living side by side, or even potentially fused together.

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OfflineMycomyth
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Re: dikaryotic mycelium questions [Re: EonTan]
    #5551355 - 04/24/06 02:55 PM (17 years, 10 months ago)

Here is probably more than you wanted to know about myc. I didn't write it, but don't know who did (the post now says "Anonymous" as the originator).
____________________
A hybrid would result from the mating of a monokaryon of one strain with a monokaryon of another strain. This paired Dikaryon would be a hybrid between the two strains. The true success of the hybrid would be if it produced fruits that had spores. Only at this stage would the true hybrid STRAIN be accomplished. No genetic information is exchanged between the two strains until their seperate haploid nuclei have fused and then undergone meiosis. Recombination would then occur between the two seperate strains forming a third strain. It's offspring(spores) would be new combinations of the two donor strains.

Hybrids can also be formed by Anastomosis between two dikaryons of different Strains, but it will happen at Far less frequency then mating monokaryons.

Anastomosis occurs at a higher frequency between substrains of an individual strain. I.E. between different spore matings resulting from a single syringe. When you multispore inoculate a jar of substrate, this is Happening with a high degreee of frequency. A dominant mating will fuse with other matings, incorporating them into it's mycelial network. It can even rewire a false mating into a good mating. It can overcome an incompatible mating, by replacing one of the nuclei within another strand with one of it's own.

A Fertile dikaryon A1B1 A2B2 can run into an infertile A1B2 A2B2 and replace the A1B2 nuclei with a A1B1 nuclei. Creating a fertile A1B1 A2B2 dikaryon that joins the colony. Subsequent fusion could replace the A2B2 nuclei with it's own A2B2 nuclei, which would completely rewire the hyphal strand to it's exact genetic makeup. Or it could leave it partially rewired.

This process can be seen on a Nutrient agar plate. Not all substrains within a strain will fuse, some are completely incompatible. There will be a zone of zero growth between them on the petriplate. They just will not fuse.

In essence hybrids can be formed between substrains of a single strain or between different strains of the same species.

This is the theory, this has been scientifically demonstrated on other species of Basidiomycetes that have been studied!!! I know of know studies that have been done on CUBENSIS, relating to this information. But it must exist. Because it has been Clearly stated in several texts, that Psilocybe cubensis is heterothallic and tetrapolar. And all the above information relates to that type of breeding system in the Basidiomycetes.

You ask why this is not being done, because not much MONEY goes into this type of research in the legitamite world.

Anastomosis has been studied extensively in the edible mushroom world. Agaricus bisporus is homothallic and two spored, not four. Each and every spore it produces already contains both haploid nuclei to make a dikaryon. But fusion(Anastamosis) between these dikaryons produces more productive Dikaryons!!! The majority of High yielding bisporous are a result of HYBRIDIZATION within a STRAIN or between Strains of this species. So if it occurs between DIKARYONS of this species,it has been viewed occuring between monokaryons of other species, WHY WOULD anyone Doubt that it can occur within the Species Psilocybe cubensis.

The major factor to overcome with mating monokaryons is the proximity with which they germinate. Spores tend to clump. So simply placing spores of two different strains in a single syringe, will not overcome the clumping of spores of like strains, and hence their close proximity to each other upon germination.

Probably still occurs, when injected into a substrate, and some of the resulting fruitbodies might actually be hybrids, between Strains, and the resulting offspring (spores) from that mushroom will be different looking then both the mushroom it came from, and all of the mushrooms from both the Donor strains.

Much easier to DILUTE spores from each strain seperately, plate them, isolate slower growing monokaryons, and try mating as many of these from each strain as possible, with as many as possible from the other strain. All matings that fruit, are hybrids!!! If they produce spores, you now have a new STRAIN. Simply cloning the original matings that fruited, will be a Hybrid as well, but not a true hybrid, becasue their has been NO Recombination between the two strains, NO MIXING OF GENES. There has simply been a successful coexistence of two haploid nuclei, one from each Strain, acting independently, but together to create fruits!!! The real genetic swap occurs during Karyogomay and the subsequent meiosis.
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InvisibleDarkenshroom
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Re: dikaryotic mycelium questions [Re: Mycomyth]
    #5552110 - 04/24/06 07:08 PM (17 years, 10 months ago)

Teonan knows what he is talking about...Teonan was the anon poster of that info, if you read the thread it came from people mention it.

Darken
*smiles*


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OfflineRogerRabbitM
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Re: dikaryotic mycelium questions [Re: Darkenshroom]
    #5553299 - 04/24/06 11:15 PM (17 years, 10 months ago)

But, I don't consider monokaryons from one strain mating with monokaryons from another strain to be 'hybrids' because they're the same species. Of course they'll pair up and the offspring is fertile. It's similar to humans from Europe mating humans from South America or Africa. The results are not hybrids, but simply 'crosses'.

A cross-species pairing would be a hybrid.
RR


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Invisiblemycogirl
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Re: dikaryotic mycelium questions [Re: RogerRabbit]
    #5553347 - 04/24/06 11:25 PM (17 years, 10 months ago)

True, but to play the devil's advocate, what defines a species? I ask because plants are weird, and don't follow animal definitions of species. And we all know fungi is way weird compared to plant or animal. So much cross over.

(P.S. no one else considers them hybrids either. I think at best they would be referred to as varieties or cultivars. )


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InvisibleRoadkillM
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Re: dikaryotic mycelium questions [Re: Darkenshroom]
    #5553499 - 04/25/06 12:05 AM (17 years, 10 months ago)

EonTan = Teonan

:smile:


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OfflineWorkmanV
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Re: dikaryotic mycelium questions [Re: mycogirl]
    #5554745 - 04/25/06 12:17 PM (17 years, 10 months ago)

I found this definition online which sounds pretty good to me:

Hybrid:

Genetics. The offspring of genetically dissimilar parents or stock, especially the offspring produced by breeding plants or animals of different varieties, species, or races.


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Invisiblemycogirl
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Re: dikaryotic mycelium questions [Re: Darkenshroom]
    #5557311 - 04/26/06 01:24 AM (17 years, 10 months ago)

Quote:

Darkenshroom said:
Teonan knows what he is talking about...Teonan was the anon poster of that info, if you read the thread it came from people mention it.

Darken
*smiles*




Thanks then Teonan...those anon posts blew my mind. I think only my myco prof knew that much out of anyone I have known...


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Re: dikaryotic mycelium questions [Re: RogerRabbit]
    #5566405 - 04/28/06 09:37 AM (17 years, 10 months ago)

Roger what is your definition of a hybrid? I have studied Plants, fungi, and animals, and a hybrid is defined within the rules of the organism you are crossing.

Since Fungi don't naturally hybridize between species, YOUR orthodox definition is not applied. Read a book on fungal genetics and they refer to outcrosses as hybrids.

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OfflineRogerRabbitM
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Re: dikaryotic mycelium questions [Re: EonTan]
    #5566499 - 04/28/06 10:17 AM (17 years, 10 months ago)

There is such little difference between strains of fungi, especially as 'strains' are referred to in the OMC, that I can't consider for example crossing a PR with an EQ and calling it a hybrid. The fruits are usually indistinguishable from each other and I can see no difference under the microscope between spores.(albino and red spores excluded) Therefore, how could it be a hybrid?

Also, fungi species have been successfully crossed, so I prefer at least for my own experiments to consider it a hybrid when that happens, and not simply crossing strains within a species. I realize I'm taking a minority position on this and most others disagree.
RR


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Re: dikaryotic mycelium questions [Re: RogerRabbit]
    #5566664 - 04/28/06 11:23 AM (17 years, 10 months ago)

There is such little difference between strains of fungi?

What the HAY are you talking about. They are different at every level except Species. Thats why they are strains.

They are different genetically, morphologically, and physiologically from one another. They just belong to the same species.

If you can't see the differences microscopically, you need to look again More carefully.
If you can't see the differences Macroscopically, then you are blind.


Yes species have been crossed in a lab. INTRASPECIFIC hybrids. Even intrageneric hybrids. But this does not do anything to argue against the intrastrain hybrid. Or does it negate the Differences between strains.

I just don't see your ARGUMENT.

How about plant pathogenic fungi, and host specificity? Strains are REAL, they have meaning beyond the naming of a Mushroom for retail sale on the internet.

Now if you did do a family tree Mating compatability study with every strain in circualtion, I think you would find that alot of them share some common ancestory, so they share some or alot with other strains, but you will also find that there are VERY REAL and DISTICNT STRAINS in circulaiton.

Breeding studies with other fungi have revealed this to be true. And have been documented in books, and journals.

Becasue the spores look similar doesn't make them the same. Every put some copelandias under a scope. Do wet mounts in water of fresh gill frags and tell me there are no differences between strains of Copelandia cyanescens. Becasue they have large easily visible Cystidia it makes for VERY EASY comparisons. Unlike most of the PSilocybe species, which have small features, lack pleurocystidia, etc...
If they were legal, Copelandia cyanescens would be the IDEAL species to work with for TEACHING Mushroom morphology to students. After looking at dozens of Copelandias from all over the world, I would say that Strains are very real things, and not the figment of the OMC imagination.

Just my two cents but I don't think you have a valid argument against the intrastrain hybrid, as being a hybrid.

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OfflineRogerRabbitM
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Re: dikaryotic mycelium questions [Re: EonTan]
    #5567059 - 04/28/06 01:45 PM (17 years, 10 months ago)

As I said, anyone is free to disagree. However, I can take a single sporeprint and get as much variation between the strains that emerge as you'll find between any of the 'marketed' strains out there. I have looked under a scope and there is very little to no discernable difference between 'strains' of cubensis. In my pictures here, you'll find lots of microscope pictures at 1000x. I challenge you to identify strain on any of them. I agree there is more variation in pans.

As far as identification of strains by their macroscopic qualities, if you can do that, you have powers none of the rest of us have. Even penis envy doesn't present the distinctive penis shape everytime. EQ isn't always large fruits, and PR isn't always hard to pin, Goliath isn't always the biggest cope, nor is it always a bitch to grow.

I don't insist on being right. As said, feel free to disagree. Personally however, I won't lay claim to creating a hybrid until I succeed in documenting a cross-species hybrid. Peace.
RR


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Re: dikaryotic mycelium questions [Re: RogerRabbit]
    #5587299 - 05/03/06 07:12 PM (17 years, 10 months ago)

You can't id to SPECIES under a scope by looking at spores at any magnification. You have to look at everything to id to species. Macro, micro, and habitat. And then it is still a pain in the arse. ID to strain is even harder, becasue it is a population of substrains that do vary from one another, but their are other POULATIONS that contain genes that cannot be found in other strains.

You cannot Isolate a GT from a PR print, that is the variation that you are missing.

Good luck with the cross species hybrid.  :thumbup:

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Re: dikaryotic mycelium questions [Re: EonTan]
    #5588374 - 05/03/06 11:30 PM (17 years, 10 months ago)

Hi everyone.
EonTan hit the source of the variation seen within strains on the head. What your seeing is the shuffling of the combinations of genes within each tetrad (set of 4 spores basically). As a simple example, let's say you have a culture of penis envy from a mating between two compatible spores only. This dikaryon (n n) contains one haploid genome (n) with allele 3 of gene x and another haploid genome (n) that has allele 7 of gene x. After the initiation of sexual development, karyogamy eventually occurs(the fusing of the two haploid genomes into a single diploid (2n) genome). Meiosis follows with each chromosome from one genome pairing with its homolog from the other genome. The chromosomes replicate once to form groups of 4 (tetrads). During each meiosis leading to the formation of 4 spores, recombination can occur between the four copies of any given chromosome. At the end, the four copies of a chromosome are segregated into one of the four spores. This occurs independently for each chromosome of the genome. So back to gene x. Millions of meioses occur in each fruiting body, so most spores will have widely different haploid genomes from the two original haploid genomes. If you cross two of the resulting spores, you could easily get dikaryons with two copies of either allele of gene x instead of one of each in the original dikaryon. In the end, a huge amount of variation can occur within a strain. The only way to eliminate this is to do at least 20 generations of two spore crosses while selecting for the desired penis envy phenotype after each cross.
Sorry if that was too long or simple.
As for Iding to species, a systematic examination and documentation of macro and microscopic features of several isolates of each strain is required. The formulation of a key would be the next step. It may end up that only genetic marker can identify strains accurately. That sounds like alot of work to me.

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OfflineRogerRabbitM
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Re: dikaryotic mycelium questions [Re: texas34]
    #5591044 - 05/04/06 06:40 PM (17 years, 10 months ago)

quote:
"Millions of meioses occur in each fruiting body, so most spores will have widely different haploid genomes from the two original haploid genomes"

I could be incorrect, but meioses is how I have explained the fact that not all clones grow out to be exactly like the flush the donor was picked from. I've seen cloned tissue many times where the mycelium sectors, and I've then fruited out the individual sections and found wildly different fruitbody presentation. In other words, more than one 'strain' active in a fruitbody. For those who are about to say 'spores on the stem germinated', it's not possible as the tissue was taken from deep inside the fruitbody under sterile conditions.
RR


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Re: dikaryotic mycelium questions [Re: RogerRabbit]
    #5592016 - 05/04/06 10:24 PM (17 years, 10 months ago)

RR,
I would not say you are incorrect. I do think it would be more accurate to call them sub-strains, as the total genome content of half of each tetrad (2 spores) is the same as the original dikaryon. If all the different strains about really are different, then you would have variation in all strains, but you would not get a B+ isolate from penis envy spores. A mating as I described earlier would not be a clone since the genomic content has changed. You get different sub-strains. If the strain you are working with has already been inbred to the point that both haploid genomes are virtually homologous to each other (obviously mating type loci could not be homologous), then recombination during meiosis would result in parental type haploid genomes and no true recombinants. So this gets back to a lively ongoing discussion in academia over the definition of species, strains, and sub-strains to which I offer no unique insight.

As for sectoring and differential FB phenotypes from transferred tissue, I would blame strain degradation or mitotic recombination. I'm not sure what the situation is in basidiomycetes, but in ascomycetes, chromosome deletions and discardings are seen when the fungi have been cultured on rich media for too long. The theory is that the fungi have such a good life in sterile culture that they toss out genes normally needed in the environment but no longer needed in culture. I would wager that this may not be the only trigger of strain degradation though.

In ascomycetes, the first character to be lost is sexual development followed by the loss of asexual fruiting bodies. This leaves conidiogenous cells on naked hyphae, i.e. a Cephalosporium/Acremonium state. Upon further degradation, asexual reproduction is completely lost and only sterile hyphae will grow. Once a character is lost, it will not return. To get around this asexual spores are stored long term in order to return to the original strain if degradation is occurring in your active cultures. Unfortunately this option is not available with mushrooms. Again, I don't know how well this applies to mushrooms, or if the theory is correct or not to begin with. However, I have seen sectoring in an ascomycete grown up from a single conidium (asexual spore). The entire mycelial mass must contain the same genetic content unless portions have had mutations occur. Your still geting fruiting bodies from the different sectors, so if strain degradation is to blame, it doesn't act in exactly the same way as in ascomycetes.

Another option, is mitotic recombination. This is seen in some fungi. It was first noticed in clonal populations of asexually reproducing species for which no sexual state is known. Analysis of the clonal populations showed that recombination was occurring even though it shouldn't be taking place. Once again, I have no idea as to whether this actually occurs in mushrooms.
T34
Note: edited for typos and a little content

Edited by texas34 (05/04/06 11:15 PM)

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OfflineRogerRabbitM
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Re: dikaryotic mycelium questions [Re: texas34]
    #5593225 - 05/05/06 08:43 AM (17 years, 10 months ago)

Thanks for the information. I've noticed the phenomena of sectoring clones with multispore inoculation, not when using an isolated dikaryotic strain. I see it also as a possibility that since so many (sub)strains are active in the same substrate, perhaps more than one substrain is active in a single fruitbody. That's just a theory on my part, but based on years of observation. Anyone care to comment on that theory?
RR


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Re: dikaryotic mycelium questions [Re: RogerRabbit]
    #5594089 - 05/05/06 01:02 PM (17 years, 10 months ago)

I think the conventional belief is that a fruit body is formed from one type of mycelia. It would be very difficult to determine if there was more than one type in a fruiting body. However, I would not dismiss your theory out of hand either. I think it's a perfectly valid theory even if it turns out to be inaccurate.
Meiosis alone is more than powerful enough to cause all of the variation you're seeing especially given the number of meioses occurring in one fruit body.
T34

Edited by texas34 (05/05/06 02:23 PM)

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