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Offlinechichas1
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Isolated strain question * 2
    #28340963 - 05/30/23 06:35 PM (7 months, 25 days ago)

Hello Shroomery.

Not long ago I started working on strain isolation but the more I investigate, the more doubts I have.

At first I thought that an isolated strain was exclusively a dikaryon but apparently this can only be obtained by crossing two spores (monokaryons) as soon as they germinate so that they do not fuse with others, via anastomosis.

When we manage to isolate a strain by agar transfers, I understand that what we obtain is a stabilized heterokaryotic mycelium with a diverse genetic variety, which, when isolated, gives us consistency in our grows. So, would each isolate of a multistrain be heterokaryotic mycelium? And if so, what happens when it initially divides into sectors? What do we call the mycelium from which we started?

Thanks for your attention.


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OfflineChamo
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Re: Isolated strain question [Re: chichas1]
    #28355946 - 06/11/23 05:41 PM (7 months, 13 days ago)

Hola chichas

I’m not sure I understand your question exactly.

A strain is the fusing of two compatible hyphae. Monokaryotic mycelium contains a single haploid (n) nucleus. Dikaryotic (n+n) mycelium is formed from fusing two compatible monokaryons or mating types, via anastamosis. The nuclei do not fuse, and heterokaryotic is just a way to say that there are two or more genetically distinct nuclei inhabiting one cell without combining. The cells continue to combine this way until they are ready for spore production, where they will combine together.

When you manage to isolate a strain what you obtain is an individual set of genetics. This removes the genetic diversity within your grow. It produces a consistent result, and allows you to choose the strain with the best genetics for your intended purposes.

Genetic variety comes once you have grown the strain to fruiting conditions, and the nuclei within the cell combine to create spores. This repeats the process, but that is why you keep a culture slant-to preserve the individual genetics you want so that you do not have to continually start over.

Sectoring is caused by strains differentiating in the agar, and is how you know there are still multiple strains in your sample. Do not start the isolating process with too many spores or you will have more strains to separate than is worth the time or space.

Buenas


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Offlinetrippleblack
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Re: Isolated strain question [Re: Chamo]
    #28355996 - 06/11/23 06:16 PM (7 months, 13 days ago)

Think I know what you mean.

I read a lot about isolation, and the most logical answer i gather is that single spore mating pairs are ideal for categorizing phenotypes for use in a academic or commercial breeding program.

Is calling a "single sector petri isolation" an isolate proper?  or should that term only be resered for mono - mono mated pairs.  I don't know of any authority who defined these terms.

I have not seen any evidence or research suggesting that single mono x mono mated isolates are more vigorous, stable, or better than "isolated sectors".


*i do think -from experience, the more genetics entangled with one single sector isolation, the less variation you'll see in a breeding project. i' make it a  point to limit the amount of spore germinations per "isolated culture".. 

I have seen in research directly stating that one single mushroom can be comprised of pairings seven or more unique nuclei.

would be nice to hear anyone else chime in if they came across anything different.


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OfflineChamo
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Re: Isolated strain question [Re: trippleblack]
    #28356113 - 06/11/23 07:15 PM (7 months, 13 days ago)

Oh now I understand the original question. Breeding is complicated and understudied. Even if you fail to achieve the desired result from combining two known dikaryotic isolates, it seems less time consuming than breeding from monokaryotic.

Is this relevant?

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4397376/


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Offlinechichas1
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Re: Isolated strain question [Re: Chamo]
    #28357299 - 06/12/23 03:56 PM (7 months, 12 days ago)

Thanks for your answers.

When we start from multispore, these spores combine (if they are compatible) to create dikaryons, but in turn, these dikaryons could be combined by anastomosis with other monokaryons that didn't mat yet, forming (n+n+n) and then (n+n+n+n...) giving place to a heterokaryotic organism. From what I have read, a dikaryon can combine only with monokaryons but not other dikaryons that are also fusing monokaryons, so we end up with mycelium that contains several cultures.

This would explain why sometimes growing from spores, we can identify different genotypes  in the same grow.



what I think that happens when we isolate from multispore myc, is that a completly isolated sector, could be heterokaryotic (n+n+n...) with nuclei perfectly compatible. The problem of these isolates would be that all those genes may switch to dominant or recessive not being that consistant than a true isolated strain (n+n) only obtained by breding monokaryons.

I am learning a lot with you and I appreciate the effort and information you share despite my way of expressing myself in English.


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OfflineChamo
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Re: Isolated strain question [Re: chichas1]
    #28357435 - 06/12/23 05:13 PM (7 months, 12 days ago)

If you are asking for research purposes, I also enjoy learning more about these things. But as far as practicality, I have never experienced or heard problems with a lack of consistency from these types of isolates, because they are true isolates.


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Offlinetrippleblack
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Re: Isolated strain question [Re: Chamo]
    #28364512 - 06/18/23 03:54 PM (7 months, 6 days ago)

Quote:

The problem of these isolates would be that all those genes may switch to dominant or recessive not being that consistant than a true isolated strain (n+n) only obtained by breding monokaryons.




i was getting at this in my post; i think this statement you just made is unproven.  You would think your statement may lean true..  and it might..  but i could not find any studies comparing multi nuclei 'isolates' vs a true mono + mono dikaryon -and i have searched. 

one thing i do scoff at is a "good clone"  -i hate clones, since they usually lack extreme variation.

i bet their are studies in chinese or korean which actually do attempt to answer these questions..  i would love to know.. would be a easy job for chat gpt5 once it's able to translate academic papers.


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OfflineTri-Polar
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Re: Isolated strain question [Re: trippleblack] * 1
    #28367911 - 06/20/23 10:40 PM (7 months, 3 days ago)

some things that may or may not be interesting tot he topic-

"In mycology, a strain refers to a specific genetic variant or isolate of a fungal species. Strains can be distinguished based on various characteristics, including morphological, physiological, and genetic traits (Muñoz et al., 2005). In the field of mycology, strains are often isolated and identified through a combination of techniques, such as morphological and physiological tests, molecular approaches, and DNA sequencing (Farahyar et al., 2020; Feau et al., 2009). Isolating and characterizing different strains is important for understanding the diversity, ecology, and pathogenicity of fungi (Couppié et al., 2019). Strains can also play a crucial role in antifungal susceptibility testing and the development of targeted therapies (Aho, 2009; Youssef et al., 2021). Additionally, strains may exhibit variations in their susceptibility to antifungal drugs, leading to the emergence of drug-resistant strains (Denning, 2022). Overall, the concept of strain in mycology is essential for studying and classifying fungal organisms."

"based on the provided references, it can be inferred that a strain is not defined solely as the dikaryotic mycelium of two monokaryotic spores. While dikaryotic mycelium is a characteristic feature of certain fungal strains, it is not the sole defining factor. Strains can be identified based on various characteristics, including morphological features, genetic traits, and specific markers such as clamp connections or spicules Chan et al. (2011)Sigler et al., 1997). Additionally, the development and growth of a strain can involve processes such as dikaryotization, karyogamy, and sporogenesis (Pillai & Krishna, n.d.). Therefore, the definition of a strain in mycology encompasses a broader range of characteristics and genetic processes beyond the dikaryotic mycelium of two monokaryotic spores."


"Based on the provided reference Rao & Niederpruem (1969), it is evident that the combination of a dikaryon through anastomosis with a monokaryon would result in a fungal entity with genetic information from three nuclei. However, whether this entity would be considered a true strain or not is not explicitly addressed in the reference. The concept of a strain in mycology typically refers to a specific genetic variant or isolate of a fungal species. While the involvement of three nuclei may introduce additional genetic diversity, further research is needed to determine whether this would be classified as a distinct strain or a different genetic variant within the same strain."

I'm a grade A idiot so its all greek to me but I got a purple belt in google-fu, sorry if those quotes are irrelevant :grin:


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OfflineChamo
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Re: Isolated strain question [Re: Tri-Polar]
    #28368284 - 06/21/23 07:13 AM (7 months, 3 days ago)

Thanks for actually using sources. I was attacked for saying the same thing. Nothing is as clearly defined as is claimed here.


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OfflineTri-Polar
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Re: Isolated strain question [Re: Chamo]
    #28368360 - 06/21/23 08:05 AM (7 months, 3 days ago)

Yeah, I dont understand it so I can't have too much of an opinion, but the second I find an AI that was trained on textbooks imma be unstoppable :cool: :rofl:


--------------------
Intro to Shroomery:raver2:The Sexy TEKs

IF I HAVE SEEN FURTHER IT IS BY STANDING UPON THE SHOULDERS OF GIANTS, BITCH.

hey fuck you dont look at me


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