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OfflineExcess Taters
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What happens when you cut a sector of agar from a clone plate? * 1
    #28255534 - 03/30/23 11:10 PM (9 months, 23 days ago)

I plan on running a perhaps, ill advised experiment, if this answer doesn't exist yet.  I'm wondering what happens genetically when we cut a sector of agar from a clone plate and make a transfer.

My understanding is that we cut the leading edge of a normal agar plate and throw it on new dish, now we've removed roughly 50% of the genetic diversity.  But when we clone a mushroom, does that apply in the same way?  Is all the mycelium from a clone, the same DNA, and cutting a sector doesn't reduce the genetic diversity?  Or is that incorrect.  I'd rather not run an unnecessary experiment.


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If you've never grown mushrooms before, here's how you start.  First, follow the Updated PF Tek, put 4 holes in jar lids not 1, and use a water tub! 
The next move is Shoebox Tek. After that you move onto grains, agar, monotubs.  Agar is easy, just do it.
Other useful links - Picture guide for how things should look and proper surface conditions guide
Growing APE or PE?  P9 pseudo casing tek


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Offlineherbstation
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Re: What happens when you cut a sector of agar from a clone plate? [Re: Excess Taters] * 1
    #28266707 - 04/07/23 10:38 AM (9 months, 16 days ago)

I've been waiting to see if someone more experienced will reply to this.

I have noticed that tiny agar slices or LC innocent spots seem to grow more organized plates than bigger chunks. I wonder if a smaller sample results in meaningfully less diversity.


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OfflineExcess Taters
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Re: What happens when you cut a sector of agar from a clone plate? [Re: herbstation] * 1
    #28266790 - 04/07/23 11:32 AM (9 months, 16 days ago)

The best info I found was here...

https://midnightmushroomco.com/blogs/myco-blog/fungi-reproduction-how-a-mushroom-is-born


Quote:

6a - Substrate inoculated with a single colony will colonize quickly and contain the same genetics throughout, it will be totally clonal. The fruits produced by this colony will all be the same genetically, cloning from any one of them will yield the same exact isolate, identical to the one in step 4.

Because the substrate is colonized by a single colony, all of the resources have been available and utilized towards its growth, and fruiting takes place more or less evenly across the entire surface, cooperatively.




I don't know that what's written here truly answers the question though.  It seems like it suggests that yes, whatever you cut from a clone is genetically the same.  Still, I'm not a genetics dude so I don't understand this stuff well enough to be completely convinced.  If anyone does, or this is somehow wrong, please let me know.


--------------------
If you've never grown mushrooms before, here's how you start.  First, follow the Updated PF Tek, put 4 holes in jar lids not 1, and use a water tub! 
The next move is Shoebox Tek. After that you move onto grains, agar, monotubs.  Agar is easy, just do it.
Other useful links - Picture guide for how things should look and proper surface conditions guide
Growing APE or PE?  P9 pseudo casing tek


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Offlineherbstation
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Re: What happens when you cut a sector of agar from a clone plate? [Re: Excess Taters]
    #28266855 - 04/07/23 12:22 PM (9 months, 16 days ago)

That article suggests that once you isolate a culture on agar it will always produce the same fruits when transferred or cloned. I think the clones in their example have 1 set of genetics because they only inoculated with 1 set.

I'm not sure about their claim that you can isolate down to a single culture with 1 transfer. It does seem intuitive, though, that most of variety gets tangled up in the middle while some of the genetics race for the edge.

I think I would rather select clones than select mycelium. You might weed out some good fruiting genetics by transferring only the most aggressive growth. I think that's why people try to isolate all the sectors, not just the most aggressive ones. That's totally a theory, though. That reminds me, I've definitely heard of a clone with multiple genetics because I've heard of people trying to separate the different sectors within one clone.

Edit: I get very different growth on different agar media. I wonder where the diversity resides in each growth pattern. Like I wonder if the thick strand growth is more or less diverse than the fluffy circular growth.



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Expanding my mind until I can join the collective


Edited by herbstation (04/07/23 12:26 PM)


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InvisibleWay
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Re: What happens when you cut a sector of agar from a clone plate? [Re: herbstation]
    #28268950 - 04/08/23 05:42 PM (9 months, 15 days ago)

From what I remember reading is a clone is not a monoculture.

The further you isolate a clone culture down via transfers, the fewer sectors that culture will have right? Well all those sectors made up that clone's genetics. From what I understand, you can affect the performance of a clone by isolating it down too far. Either in a good way or bad way.


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OfflineExcess Taters
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Re: What happens when you cut a sector of agar from a clone plate? [Re: Way]
    #28268979 - 04/08/23 05:55 PM (9 months, 15 days ago)

Thanks, your phrasing helped me find more info.  Had to go back and find old RR posts, man, what a legend.  Still has the answers even now.

TLDR: A mushroom may be a combo of up to 200 organisms, so you may have up to 200 sets of genetics in a single fruiting body.  So you can further isolate down a clone, though at some point it does add more problems than benefits. 

Quote:

archivist said:
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?




Quote:

RogerRabbit said:
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




Quote:

RogerRabbit said:
I've never seen sectoring from a clone that was grown from an isolated strain.  I have however seen it many times with multispore inoculated fruits.  I've also seen it with outdoor cloned fruits from many species.  Not all strains that emerge are going to be compatible, and some will remain entwined within the mycelium network, but as separate organisms.

My theory is that more than one of these strains can be present in a single fruit body.  The main evidence is that if you grow out each of the sectors from the cloned fruit, they'll in turn produce fruits that are noticeably distinct from each other.
RR




Quote:

RogerRabbit said:
It's been shown from DNA testing that up to 200 organisms can be active in a single fruiting body.  If you're growing from multispore inoculation, several substrains can be active in a fruit.  If you're growing from a single sector isolate proved on agar in a petri dish, your clones will be isolated strains, but further down the cell division pool than the original petri dish, which is what you'd want to use for further growing.
RR




--------------------
If you've never grown mushrooms before, here's how you start.  First, follow the Updated PF Tek, put 4 holes in jar lids not 1, and use a water tub! 
The next move is Shoebox Tek. After that you move onto grains, agar, monotubs.  Agar is easy, just do it.
Other useful links - Picture guide for how things should look and proper surface conditions guide
Growing APE or PE?  P9 pseudo casing tek


Edited by Excess Taters (04/08/23 06:06 PM)


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OfflineEclipse3130
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Re: What happens when you cut a sector of agar from a clone plate? [Re: Excess Taters] * 1
    #28269727 - 04/09/23 09:12 AM (9 months, 14 days ago)

If you have spores germinated and you grab a sector, sure 50% of the genetic diversity is reduced, or more.

Each time you isolate a sector more genetic diversity is reduced, say you transfer 3 times from spore and you have something growing out uniformly, there's still a lot of strains within the isolation even though it looks to be a uniform mycelium.

When you clone something from growing that out, it reduces it again even further.

At that point, the genetic diversity is so reduced taking transfers from the clone on agar won't change your outcome noticably, unless it's showing different growth patterns. As well as each time you clone again the change will continue to be reduced greatly.

I'm not sure if that helped answer your question


--------------------
"In The Material World One seeks retirement and grows Old
In The Magical World One seeks Enlightenment and grows Wiser
In The Miraculous World One seeks nothing and grows Lighter
As we all tread the Homeward Path we will explore many Realms
And one day... we will all Realize that all experiences are Simply
Different ways in which The
All-That Is
Perceives Itself"


Edited by Eclipse3130 (04/09/23 09:20 AM)


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