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Psilocybe (Agaricales) life cycle discussions: karyogamy, meiosis, anastomosis, etc * 4
    #27395480 - 07/20/21 05:26 PM (2 years, 6 months ago)

This is the place where we can get together to discuss the  underlying complexity of the Psilocybe life cycle: from monokaryon to basidia.


If you're interested in a obtaining a deeper understanding of what's going on behind the scenes with cultivation this is the place for you.

Agaricales


Glossary of Mycology terms












Edited by Stipe-n Cap (07/24/21 09:14 PM)


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: Stipe-n Cap]
    #27395497 - 07/20/21 05:38 PM (2 years, 6 months ago)

Thanks for making this thread dude.

I was curious about the sharing of genetic material between hyphae. I know anastomosis leads to fusion of cells that then contain two haploid nuclei with these haploids fusing when spores are produced. I know nuclei can be transferred between cells of the same dikaryotic hyphae. The question is: is genetic material shared between different dikaryotes? My conception of the matter is that no, it is not but when fruiting bodies begin to form various compatible dikaryotic hyphae combine and this is how strain crossing works when myceliated grain is mixed ie dikaryotic hyphae can grow over each other and might intertwine but they will not meaningfully combine until the tertiary mycelium of a mushroom begins to be created.

Can anyone offer some insight into whether or not this line of thinking is valid?


Edited by multifractal (07/20/21 06:43 PM)


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: multifractal]
    #27395524 - 07/20/21 06:00 PM (2 years, 6 months ago)

I'm having trouble finding a source for this atm, but I believe clamp connections are formed not only along individual dikaryotic filaments to exchange nuclei backwards from the terminal tip but also between interconnected filaments which form the mycelium, as you mentioned, this is how crosses would be made without single spore crossing, and then ultimately  forming basidiocarp.

I'll start digging through my shit lol, unless some other member joins in who has the source on hand.


Edited by Stipe-n Cap (07/20/21 06:38 PM)


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: Stipe-n Cap]
    #27395530 - 07/20/21 06:07 PM (2 years, 6 months ago)

I also am having a hard time finding clear answers to this, but I appreciate you helping me out in digging up some information - thats a good place to start.


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: multifractal]
    #27395542 - 07/20/21 06:19 PM (2 years, 6 months ago)

Quote:

multifractal said:
I also am having a hard time finding clear answers to this, but I appreciate you helping me out in digging up some information - thats a good place to start.




When a mushroom forms it already has the genetic information to do so, meaning the dikaryotic mycelium successfully mated as far as I understand. It can be viewed pairing on agar pre fruit body formation, though mating dikaryotic mycelium is a much lesser chance than mating monokaryotic


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: Stipe-n Cap]
    #27395568 - 07/20/21 06:43 PM (2 years, 6 months ago)

Quote:

p9hu7 said:
I'm having trouble finding a source for this atm, but I believe clamp connections are formed not only along individual dikaryotic filaments to exchange nuclei backwards from the terminal tip but also between interconnected filaments which form the mycelium, as you mentioned, this is how crosses would be made without single spore crossing, and then ultimately  forming basidiocarp.

I'll start digging through my shit lol, unless some other member joins in who has the source on hand.



Remind me to dig up what I have


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: Eclipse3130]
    #27395570 - 07/20/21 06:46 PM (2 years, 6 months ago)

I can find nothing to support that dikaryotic hyphae "mate", meaning to exchange/combine in some fashion genetic material, with other dikaryotic hyphae - before fruit formation or otherwise.

Interesting image showing the expression of DNA from two parent haploids that I posted in another thread:



Edited by multifractal (07/20/21 06:57 PM)


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: bodhisatta]
    #27395573 - 07/20/21 06:47 PM (2 years, 6 months ago)

Quote:

bodhisatta said:

Remind me to dig up what I have




You are officially reminded :lol:


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: multifractal]
    #27395582 - 07/20/21 06:53 PM (2 years, 6 months ago)

I've been studying along these lines too, so thread is a welcome sight. Seems that some of the wisdom from the old heads isn't making it to the next generation, and i've been wondering where to start with sharing what i know. Thanks for this.

Just a little background summary for what's being discussed here in case anyone is curious and feels left out (from THIS THREAD):

Quote:

Workman said:
Spores, like eggs and sperm, have only half the chromosomes of somatic cells. A singe spore germinates and grows a thin monokaryotic mycelium until it comes in contact with another strand of mycelium from a different spore. At the point of contact the two myceliums fuse and genes recombine into a dikaryotic mycelium that grows thicker and faster and is "hopefully" capable of fruiting.

The difficult part in breeding is isolating individual spores and growing out the monokaryotic mycelium. "The Mushroom Cultivator" (Stamets and Chilton) covers the spore diltution technique on pages 340-341.

There is another method of crossing strains that isn't as well understood or well known called anastomosis. This is mentioned on page 8 of "The Mushroom Cultivator". Anastomosis is where two dikaryotic myceliums fuse, exchange genetic material and form a new strain. This can sometimes be seen in casings containing two different strains where a few mushrooms seem to be intermediate between the two parent strains. Anastomosis can be done easily on agar where the two different fruiting strains are allowed to grow together in a single petri dish. Typically, a zone of incompatibility forms where the two strains meet. Even though it seems that the two strains are completely rejecting each other, genetic exchange is usually taking place. If a small wedge is taken from the incompatibility zone and culture out to fruiting, new strains often result mixed in with the parent strains. For some reason the crosses appear more abundantly in later flushes. It is suggested that strains very different in appearance are chosen for crossing by this method so that they are easily recognized when they occur.





this thread contains some wisdom for you. Workman is a professional that was active here up until several a FEW years ago. You can search his posts through the Advanced Search Function. Ive actually made custom search keywords in firefox for trusted (actual, professional, trained) mycologists ( like Workman, Alan Rockefeller, John Allen, etc. ) but that's not to say that everyone else doesn't add value, this is just where i go for individuals that i trust have done the work and followed a semi-academic path in doing so.

Quote:


Workman:
Also, there is no genetic recombination until just prior to spore formation.  Each cell (generally) contains a distinct nucleus contributed from each parent.  These don't fuse and get recombined until new spores are formed.  It is a minor point and doesn't change Aero's overall excellent post. 





edited: several


Edited by mycosavant (07/20/21 07:03 PM)


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: multifractal]
    #27395611 - 07/20/21 07:16 PM (2 years, 6 months ago)

Quote:

multifractal said:
I can find nothing to support that dikaryotic hyphae "mate", meaning to exchange/combine in some fashion genetic material, with other dikaryotic hyphae - before fruit formation or otherwise.

Interesting image showing the expression of DNA from two parent haploids that I posted in another thread:






I'll dig through my books tomorrow, some of this is difficult to track down via Google on my phone.

Here we go:

"A dikaryon can no longer accept other nuclei, but it can still donate nuclei to a monokaryon, a phenomenon called the ‘Buller phenomenon’."

https://royalsocietypublishing.org/doi/10.1098/rspb.2010.1110


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: Stipe-n Cap]
    #27395630 - 07/20/21 07:26 PM (2 years, 6 months ago)

Quote:

Anastomosis is where two dikaryotic myceliums fuse, exchange genetic material and form a new strain.





Again, everything I could find only mentions anastomosis occurring between monokaryotic hyphae when it even specifies the type of hyphae.

It's strange how little I could find on this. I also find it odd that so many posts on this site refer to anastomosis as mating of dikaryons; I wonder where this information originated.

Any textbooks on mycology you guys would recommend? My old microbiology textbooks touch on fungal reproduction here and there but it's pretty surface level stuff.


Edited by multifractal (07/20/21 07:29 PM)


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: mycosavant]
    #27395638 - 07/20/21 07:29 PM (2 years, 6 months ago)

Also, this is from the "General Mushroom FAQ (outdated)"... this info is not outdated from everything i've come to understand through careful observation and literature studies.. I was going to quote from it, but after reading it again, decided to post it in its entirety since it's pretty meaty and within this topic.


Quote:


Teonan said:

Q: Is it possible to cross 2 strains?
A:
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.






This GEM: Cubensis Lifecycle Questions from 4 years ago has too much to quote. It's hitting all around this thread's topic and what's been discussed thus far.



Edited by mycosavant (07/20/21 07:53 PM)


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: multifractal]
    #27395671 - 07/20/21 07:47 PM (2 years, 6 months ago)

From everything I've read there's only 3 instances where fusion occurs:

1. When monokaryons undergo plasmogamy and become dikaryotic;

2. Buller phenomenon: when dikaryotic hyphae form new clamp connections to other compatible monokaryons to exchange nuclei as they grow;

3. When karyogomy fuses two nuclei to form
diploid zygote.

When fruiting: many dikaryotic strains weave together forming fruitbodies, their terminal tips swell and form basidia where karyogomy takes place and spores ultimately form.






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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: Stipe-n Cap]
    #27395704 - 07/20/21 08:16 PM (2 years, 6 months ago)

"buller phenomenon" interesting, I've never heard of this, have you?  This opens up some good reading opportunities. Thank you again for spending time on this one, I really appreciate and recognize your dedication to what a lot of people would call a beside-the-point topic. It's a silly tag that usually doesn't mean much to me but I'm glad you're a TC.

Excited to see if others (somebody remember to remind Bod for God's sake) have anything to say as well.


Edited by multifractal (07/20/21 08:22 PM)


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: Stipe-n Cap]
    #27395707 - 07/20/21 08:18 PM (2 years, 6 months ago)

Yep. Ps. cubensis is a heterothallic and tetrapolar basidiomycete.

Somer pertinent definitions:

Merriam-Webster Dictionary says:
Quote:


Heterothalic adj. -
het·​ero·​thal·​lic | \ he-tə-rō-ˈtha-lik

Definition of heterothallic

1 : having two or more morphologically similar haploid phases or types of which individuals from the same type are mutually sterile but individuals from different types are cross-fertile
///heterothallic fungi
///heterothallic spores


---

tetrapolar adj -
tet·​ra·​polar | \ ¦te‧trə+ \

Definition of tetrapolar

: having four poles
///certain abnormal mitotic figures are tetrapolar






From Wikipedia:
Quote:

Heterothallism is the most common mating system in Basidiomycota and in Agaricomycotina (the mushroom-forming Fungi) about 90% of the species are heterothallic.[19] The tetrapolar type of mating system is ruled by two unlinked mating loci termed A and B (in Agaricomycotina) or b and a (in Ustilaginomycotina and Pucciniomycotina), both of which can be multiallelic. The combination of A and B (or b and a) alleles, termed mating type, determine the specificity or sexual identity of the individual harboring them. Only individuals with different specificities are compatible with each other and therefore able to start the mating event. A successful mating interaction begins with nuclear exchange and nuclear migration resulting in the formation of dikaryotic hyphae (containing separate haploid nuclei from both initial parents). Dikaryotic hyphae, under the appropriate environmental conditions will give rise to the fruiting body which contains the basidia – specialized cells in which sexual recombination via karyogamy and meiosis occurs. This dikaryotic condition in Basidiomycota is often maintained by a specialized hyphal structure called a clamp connection. The formation of clamp connections is regulated by both mating loci. Examples of tetrapolar organisms are the smuts Ustilago maydis and U. longissima,[20][21] and the mushrooms Coprinopsis cinerea, Schizophyllum commune, Pleurotus djamor and Laccaria bicolor.[22] It is believed that multi-allelic systems favor outcrossing in Basidiomycota. For example, in the case of U. maydis, which bears more than 25 b but only 2 a specificities, an individual has an approximately 50% chance to encounter a compatible mate in nature.[23] However, species such as C. cinerea, which has more than 240 A and B specificities each, and S. commune, which has more than 339 A specificities and 64 B specificities, approach close to 100% chance of encountering a compatible partner in nature due to the huge number of mating types generated by these systems.[24]




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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: multifractal]
    #27395708 - 07/20/21 08:19 PM (2 years, 6 months ago)

I love all things mycology, it's quite enjoyable to me. Thanks for that compliment, I do appreciate it.

@mycosavant

Great info there.


Edited by Stipe-n Cap (07/20/21 08:28 PM)


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: Stipe-n Cap]
    #27395729 - 07/20/21 08:36 PM (2 years, 6 months ago)

I'm also glad we're clearing up the definition of anastasmosis when applied to mycology, which is just a general term of cell fusion that could take place between hyphae within the same network (I believe this is correct. Meaning that nutrients and nuclei can be shared between dikaryotic hyphae originating from the same parent haploid pairing and not between distinct dikaryotic hyphae as would be assumed in the use of the word in regards to "dikaryon mating") as well as between monokaryotic hyphae in order for plasmogamy to occur.


Edited by multifractal (07/20/21 08:49 PM)


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: multifractal]
    #27395739 - 07/20/21 08:41 PM (2 years, 6 months ago)

Pretty much:

"In basidiomycetes, as in other filamentous fungi, syngamy means 'hyphal fusion or anastomosis' "


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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: mycosavant]
    #27395762 - 07/20/21 08:59 PM (2 years, 6 months ago)

Something of interest in terms of the lifecycle of Ps. cubensis. It appears based on early laboratory research that the life cycle of the species is determined by the environment in which it is found natively. It's worth noting that they're working with specimen that are very close to their wild genetics, so It's unclear how much of an effect the domestication and "stabilization" of varieties has on this characteristic. I just thought i'd throw this in here while i'm at it.

Full paper can be found here 

This is from a 1976 paper entitled:

PSILOCYBE CUBENSIS (AGARICALES):
A COMPARISON OF MEXICAN AND
TEXAS TYPES


Jackson, R. E., & Alexopoulos, C. J. (1976).
Psilocybe cubensis (Agaricales): A Comparison of Mexican and Texas Types.
The Southwestern Naturalist, 21(2), 227. doi:10.2307/3669959


Quote:


CONCLUSIONS.  Texas P. cubensis has a shorter life cycle in vitro
than the Mexican P. cubensis. The adaptive value of a shorter life
cycle in an area where long periods of drought and severe winters are prevalent is clear. In Chiapas, the climate is such that somatic
growth is favorable: Palenque receives twice as much rain and temperatures never reach freezing. In our laboratory both the Texas and Mexican types delayed fruiting until conditions were made slightly unfavorable for somatic growth by cutting agar strength in half. Further studies are being conducted on specific nutrient requirements of P. cubensis




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Re: Basidiomycetes life cycle discussions: karyogamy, meiosis, anastomosis, etc [Re: mycosavant]
    #27395803 - 07/20/21 09:32 PM (2 years, 6 months ago)

Cool paper, thanks for sharing. It's rare to see articles specific to P. cubensis.


Edited by multifractal (07/20/21 09:33 PM)


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