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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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Questions about strain isolation
    #9557988 -

Hey all.  After watching RR's video on strain isolation and doing some reading on the forums on the topic, I was still confused about a few details so I wanted to make sure my understanding of it was correct.  Mods, feel free to move this to general cultivation if this doesn't belong here, but I thought I'd get some higher quality answers in this forum.

1. When you create a monoculture after repeat transfers from sectoring mycelium, you isolate single sets of genetics that you then fruit individually.  Finding one with desirable attributes and then going back to that master culture allows you to grow essentially the "same" mushrooms every time.  Is that correct?

2. Suppose you clone a fruit produced from that monoculture.  Are the genetics similarly preserved?  I.e. is it the same as going back to the original master or have the genetics differentiated by then?

3. What about taking a spore print from a fruit produced from that monoculture?  As you then essentially starting over because the print is a "multispore," or are you still preserving the superior genetics of the strain you originally isolated to any extent?

4. What if, instead of starting with isolation, you grow from a multispore inoculation.  When you take a resulting fruit with desireable attributes and clone it, are you then creating a monoculture?  I have read a few posts that suggests that even within the same fruit, there can be several varying genetic strains, so my guess is no.

5. Lastly, suppose you then isolate a handful of monocultures from that MS tissue culture.  Are you then eliminating the non-fruiting strains since the original culture was from fruiting mycelium?


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Proud supporter of the canning jar industry.

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Re: Questions about strain isolation [Re: archivist]
    #9560970 -

1)  Yes.

2)  Yes, but you wouldn't want to clone, you'd want to go back to your culture slant(s) and grab the original.  I doubt senescence would have set in after one grow, but may as well use the culture with the least number of cell divisions.

3)  You're preserving the genetics to some extent, but by taking a sporeprint, you'd not be getting an exact duplicate.

4)  I've had clones from multispore inoculation that were monocultures, and I've had some that sectored.  Sectoring is especially common with strains isolated from the wild.  Other growers have theorized that these were not sectors but different mycelium types, but they were actually distinct sectors.  Stamets has confirmed this.  I've heard him say that up to a hundred or more strains and/or species of fungi and bacteria can dwell within a single mushroom.

5) Beats me. :shrug:
RR


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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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Re: Questions about strain isolation [Re: RogerRabbit]
    #9561223 -

Thanks RR, that really helped clear it up for me. 

I wanted to make sure I've understood the concept of senescence.  Just let me know if I have this straight:

Doing transfers on agar or via g2g perpetuates the same generation of mycelium, as does cloning the fruitbody produced by the same mycelium.  Senescence begins to set in after multiple transfers or multiple clones across bulk media (i.e. too much cell division has occurred), resulting in slowed growth, which is why it is ideal to go back to the youngest mycelium from that generation -- the master culture.

When you take a print from a fruiting mushroom from that generation and grow that out, does that mean everything "resets" from a senescence perspective, or have the genetics degraded somehow from the previous generation?

In the realm of psychoactive mushrooms, is it safe to say that a generation that has succumbed to senescence will produce fewer actives, or is this still inconclusive?


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Proud supporter of the canning jar industry.

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Re: Questions about strain isolation [Re: archivist]
    #9563315 -

You've got the concept down. As for senescense being reset, RR feel free to correct me, but I believe that MS can help the situation, but unless you cross it with spores from another strain you will eventually get genetic decline...

And with the actives (or any others for that matter), you will see senescence express itself in different ways, more and more over generations. Mutations, gills growing on tops of caps, lower yields, ect... not necessarily lower yields in the begining, but eventually...


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EmperoR ReishI

Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]

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Re: Questions about strain isolation [Re: RogerRabbit]
    #9563324 -

RogerRabbit said:


2)  Yes, but you wouldn't want to clone, you'd want to go back to your culture slant(s) and grab the original.  I doubt senescence would have set in after one grow, but may as well use the culture with the least number of cell divisions.




RR
  think I understand the definition but just to clarify how to use p-values:
  generation 1 from spore is p1
  a subculture of p1 is p2 
  will a clone from a mushroom growing from a p2 culture  also have p2 value since its from the same mycellium ?
  when cloning from field specimens since we don't know how many generations the mycellium has gone thru we can't attribute a p-value.,..correct?
thanks sromes1

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Re: Questions about strain isolation [Re: sromes1]
    #9564056 -

Im not RR but can answer your question.  P-values stand for the number of plates it has grown(as u seem to understand).  say you use 110mm plates.  well this is your standard then.  so a p-2 would have grown ~55mm, the radius of 1 plate.  now why would be a p-1...usually one does not let a spore germination grow that far out.  so P-1 is the 0 mark (for me anywho).  so your p-values depend on the size of your plate. 

Your clone from a p-2 plate would not have the same value.  it has probably traveled 100's of plates worth of cell growth in your bag/jar etc...  this is one huge advantage to the person working with agar!

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Re: Questions about strain isolation [Re: mycelialfruit]
    #9564182 -

I dunno if your question was about p-values in general or the stamet's p-value system which is a bit more precise (but basically the same)

Here's a link to stamet's p-value system

http://books.google.com/books?id=y1fnacRg1AYC&pg=PA99&lpg=PA99&dq=paul+stamet+p-value&source=web&ots=VOCK0V5qoO&sig=h1bdDjYCpkaJLPxc6tiTVZe0Hb0&hl=en&sa=X&oi=book_result&resnum=1&ct=result#PPA99,M1


--------------------
EmperoR ReishI

Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]

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Re: Questions about strain isolation [Re: Emperor Reishi]
    #9624442 -

So, I couldn't find any topics which mentioned this seemingly simple question:

What is the easiest way to fruit out your prospective isolates...

Do you guys do like, 1 cake per isolate? Or just go about your grows trying a different one every time?  Other than edibles, if you were a serious mycologist, it seems like you'd have more mushrooms than there'd be use for just trying to isolate a single species doing fullscale grows.


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Re: Questions about strain isolation [Re: RedBeerd]
    #9625501 -

You would want to do mini-trials with the same exact substrate formula, and environmental variables you will be growing the desirable strain on in the future, except using maybe a smaller tray size, hence "mini-trials". I usually do about 3-5 trays per isolate.

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