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Getting a strain isolate without agar ??
    #1304592 -

one of the alternatives to cloning wiith agar is doing a couple g - g transfers, which I've read allows the stronger, more rhizomorphic mycelleum to dominate.....so with this in mind....say you colonize some WBS....there are noticable differences in mycelleum quality throught the cake surface (from multiple spore germination)....can you cut out pieces that appear to have strong rhizo growth and use them to innoculate spawn?
Anyone tried anything like this before?


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There are more important things in life than just being really, really, ridiculously good looking. -Zoolander

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Re: Getting a strain isolate without agar ?? [Re: majik_monkey]
    #1304612 -

yes, it will selectively weed out some of the weaker breeding strains. It wont be as efficient as agar, and I wouldn't go past 3 generations of transfers. But it should work as a course prefilter, at least.


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Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--

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Re: Getting a strain isolate without agar ?? [Re: majik_monkey]
    #1304646 -

As RAADT said so masterfully .....YES is possible
Risky...contam risk is greater , you must man handle grain alot to harvest selected colonies .
Tough...may be small clumps you harvest to do transfer with , thus making agar much more controlled IMHO . If colony you choose is small , may not be quite enough to do "good" transfer . What safe ratio is aprox 25% in G2G transfers ?
But very possible with TLC !

oopppsss sorry brain farted this was using cakes to take isolation from ....anywho good stuff !


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Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !

Edited by tripndicular (02/13/03 04:45 AM)

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Re: Getting a strain isolate without agar ?? [Re: tripndicular]
    #1304968 -

i feel the vendors are shiping very good "isolates" as is....they prity much do all the work for you by sending you the spores of the strains they isolate.


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work of fiction

http://www.thenook.org/

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Re: Getting a strain isolate without agar ?? [Re: IDontGrow]
    #1305015 -

spores will germinate and create their own strains, the vendor isolation has nothing to do with anything. Vendors or the original person, can domesticate the strain some, by environmental stressing, and over time the genetics may change. But when spores germinate they each create a dikaryotic strain of mycellium, doesn't matter if it's spores from a cow patty, or from a syringe.


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Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--

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Re: Getting a strain isolate without agar ?? [Re: Raadt]
    #1305070 -

Thanx RAADT , I was about to go into each spore has its own genetic make up , so issolations are still required , if that is the sort of thing you are into .
I love doing agar isolates , almost as much as eating the little fellas !


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Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !

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Re: Getting a strain isolate without agar ?? [Re: majik_monkey]
    #1306390 -

Select a mushroom that shows good characteristics. (large, healthy, fast growing, etc)

Make your grain jars as usual (maybe a little less water).

After pressure cooking and cooling, use a sterile syringe to suck up some hydrogen peroxide (3%) and inject about 5mL (mL = cc) into each jar. Shake the jars well.

Use a sterile scalpel to cut pieces from the inside of the mushroom's stem.

Place these pieces into the jars and shake well.

Colonize as usual.

You have now created a fruiting isolate a.k.a. clone! Case or spawn as usual.

This worked well for some people I know. Thanks to Rush Wayne, Mycofile, Anno and others for helping people with these ideas.


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"The real voyage of discovery consists not in seeking new landscapes but in having new eyes."
- Marcel Proust

I wish you all ceaselessly flowing moments of happiness.

Edited by psyphon (02/13/03 05:02 PM)

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Re: Getting a strain isolate without agar ?? [Re: Raadt]
    #1306430 -

I don't completely agree.

With each clone/isolation you are creating a bottleneck event, thereby reducing the available genetic diversity.

Joshua


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The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1307634 -

In the spores?


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Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--

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Re: Getting a strain isolate without agar ?? [Re: Raadt]
    #1307853 -

I would have though if you fruited an isulate in a casing that all the mycelium would hold the same genetics there for semi stabalising its spores in its fruits due to lack of other germinated spores being present though the isulate will contain differing genes from the origanal multispore you isulted from and if you continued the process would it not be possible to stabalise the strain making even multi spore inoculations stable?


newbie question  :crazy:


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er

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1307912 -

Josh, what do you mean by bottleneck event? :confused:

And if it reduces the genetic diversity, isn't that good?  Cus the reason you are cloning is to reduce the diversity and have it more or less the same so that the fruits should resemble the parent.

Just curious... :tongue:


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"Know your Body - Know your Mind - Know your Substance - Know your Source.

Lest we forget. "

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Re: Getting a strain isolate without agar ?? [Re: Azmodeus]
    #1307927 -

bottleneck event - you're taking the spores only from the best shrooms, selectively excluding all but the best genetics...


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"A celibate clergy is an especially good idea, because it tends to suppress any hereditary propensity toward fanaticism." - Carl Sagan

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Re: Getting a strain isolate without agar ?? [Re: the spiral]
    #1307956 -

bottleneck event - you're taking the spores only from the best shrooms, selectively excluding all but the best genetics...

But weren't we talking about taking a tissue sample from the most charismatic, not spores?
I assume once the mycelium from the tissue fruited you wouldn't harvest those spores but start again from a multispore then pick the best.  The flush from the tissue sample will have all fruits similar, but spores aren't good is this correct? :confused:


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"Know your Body - Know your Mind - Know your Substance - Know your Source.

Lest we forget. "

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1308436 -

Well put Joshua!
Isolating without agar will be so much more difficult because it will be hard to see.


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Re: Getting a strain isolate without agar ?? [Re: G a n j a]
    #1308444 -

I dont understand your post.


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Re: Getting a strain isolate without agar ?? [Re: G a n j a]
    #1309001 -

Spores are haploid. This means they carry 1 set of genes.

When spores germinate they produce haploid hyphae. Hyphae are the filaments of cells that make up mycelium. These are labeled + and -. + and - are called mating types. When a haploid hyphae of one mating type crosses path with the opposite mating type. The hyphae fuse and undergo a process called plasmogamy. Plasmogamy is the fusion of the cytoplasm between cells. Once plasmogamy has occured the nuclei fuse and combine chromosomes to form a diploid nuclei. Diploid means the nucleus has two copies of each gene. One copy came from each mating type.

The new diploid cell then divides to form additional diploid cells. It is usually the diploid mycelium that grows faster. When the correct environmental queues are available the diploid mycelium forms a small mass. The is mass is a hyphal knot (from hyphae) and eventually grows into a complete mushroom.

Inside the gills of the mushroom are the terminal ends of the dikaryotic mycelial cells, it is at these cells that karyogamy occurs to form the diploid basidium. The diploid basidium then undergo meiosis to form four haploid nuclei. Each basidium also produces an appendage through which the haploid nucleus travels to form the basidiospore. These basidiospores then break free and are held between the gills by an electric static force. When the conditions are right the haploid basidiospore is released and carried away by air currents to germinate into new mycelium on another substrate.

______________________________________________

Each fruit is made up of a congregation of hyphae. These hyphae have two copies of each gene just like you and I (most of us at least). When you clone a fruit you are selecting a specific set of genes. These genes are present twice with in the culture. When you fruit this clone the spores from it will only be rearrangements of these two sets of genes.

A bottleneck event is an event in which the genetics of a population is reduced greatly. Imagine 100 people are left on the Earth. Only the genes from these 100 people will be present in future generations. If all of these people were Eskimos, the future progeny would appear as Eskimos. You would lose all other races.

When you isolate or clone you are causing the above circumstance. You are allowing only a small number of genes to be passed on to the next flush.

At least that's how I understand the basics. Keep in mind that new genes can arrive through mutation, but the rate of mutation is usually very low. Chromosomes can also undergo crossovers and square dancing....but that's a whole nother story.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1309009 -

>When you isolate or clone you are causing the above circumstance. You are
>allowing only a small number of genes to be passed on to the next flush.

But, Joshua, you are making this very thing happen also if you give me a print of your mushrooms.
You give me 1 print of 1 mushroom, same as if you had given me 1 clone of 1 mushroom and I grew it out and made prints of this.

Wher?s the difference?


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Getting a strain isolate without agar ?? [Re: Anno]
    #1309013 -

Correct me if I'm wrong here, but mushrooms are made of condensed mycelium. If you clone a mushroom you are only getting the genes from that shroom. If you take a print each spore has different genes. Each spore emits a hypha which branches off and forms a primary haploid mycellium. It eventually meets up with another compatible haploid mycelium and their cell walls break down and they exchange genetic information. Thus a secondary diploid mycelium if formed which is capable of producing fruits.
It would be like the difference of taking a skin sample from me which would contain my exact genetic make up or a semen sample which does only contain the limited amount of genes I could pass on but still has many more genetic variations.
I didn't read all the posts in this thread just the one I replied too, so I could have missed the point entirely now that I think about it. If you're talking about printing the clone or printing a shroom grown from a print. I suppose it's getting to the point where you can just ignore my worthless babbling. It's 4:20 in the morning and I'm sick. Sorry for the inconvenience. You're the man Anno.


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Think for yourself; Question Authority

Edited by TeRzMaStA (02/14/03 11:15 PM)

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Re: Getting a strain isolate without agar ?? [Re: Anno]
    #1309026 -

In the isolation process I have isolated a small fraction of the original genes in the multispore inoculation. The fruits from this isolation will have fewer genes than were present in the original group of spores. With each consecutive isolation/clone you narrow the available gene pool from the print you started with.

In the wild spores from different fruit bodies will mix to propogate genetic variability.

If I gave you a print and you fruited from it and took a single print from that flush and continually repeated this proceedure you would find your genenic variability to be reduced with each consecutive print.

Lets say you started w/ 100,000 available variations of a particular gene. Each variation is present once in each of 100,000 spores you germinate. You germinate these spores at a single point. From this point you sector 25% of the growth, which should contain 25% of the original variation, 25,000 variations. If you then sector 25% of this growth you are down to 6.25% of your original variation w/ 6250 variations.

I'm sure my #'s aren't accurate, but the principle should be sound.

What are your thoughts?

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1309040 -

>The fruits from this isolation will have fewer genes than were present in the original group of spores.

The fruits from an isolation are all are genetically identical (if you assume there are no mutations occurring underway), they have the same genes as the mushroom they originated from.

So again:

I harvest one particular mushroom, take a print from it, and clone it.

I then grow this clone out, and bring it to fruit and take a print again from this cloned mushroom.

What I?m saying is: Both prints will be identical.

So why should cloning be a bottleneck event?
It?s not more bottleneck than if I buy a print from a single mushroom.
And if you buy a print, then it?s mostly of one single specimen.


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Getting a strain isolate without agar ?? [Re: Anno]
    #1309066 -

Post deleted by administrator.

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Re: Getting a strain isolate without agar ?? [Re: Anonymous]
    #1309076 -

And? Same with the print of the cloned mushroom.


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Getting a strain isolate without agar ?? [Re: Anno]
    #1309191 -

Yes, I understand what you are saying.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1309200 -

I think we need Teonan here.....

Teonan ! !


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1309888 -

Josh,

Just a minor point but, the 2 nuclei do not fuse (karyogamy)after plasmogamy, but continue growth as a dikaryotic, binucleate organism.  Division at this stage usually proceeds by clamp connections.  As you said later in your post, it's not until the gills are formed that certain dikaryotic cells (the
developing basidia) undergo karyogamy to create diploid cells.  Very shortly after this meiosis occurs and haploid spores are generated from those cells.  The diploid phase in basidiomycetes is very short and only occurs in the spore bearing part of the fruit body.  The faster growing mycelium is dikaryotic and not diploid.  Good post though :smile: .

Peace

DH 

Edited by DethHed (02/15/03 11:49 AM)

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1309922 -

Exalent reading thanks for taking the time :smile: very good thread.


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er

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Re: Getting a strain isolate without agar ?? [Re: spores]
    #1310104 -

Good point. I was confused about the term dikaryotic v. diploid. You have cleared this up for me.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: TeRzMaStA]
    #1310167 -

Quote:
It would be like the difference of taking a skin sample from me which would contain my exact genetic make up or a semen sample which does only contain the limited amount of genes I could pass on but still has many more genetic variations.



Not totally sure how this works in mushrooms, but for humans, you are mistaken. A DNA sample from a skin cell contains 100% of your genetic information. A semen cell (or a female's egg cells) contains only 1/2 of your genetic information, a random selection from from each pair. A complete semen sample could contain your complete genetic makeup, but each cell would still only contain 1/2. A semen sample will never contain more genetic variety then your DNA because you cannot produce additional genes. This is why inbreeding leads to deformed children, the lack of genetic variation.
I hope that was clear.  :smile:

jssmthrfcknchrst 


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Re: Getting a strain isolate without agar ?? [Re: Anno]
    #1310254 -

I think Joshua's point was that the continued isolation or cloning of the mushroom whereby the same exact genetic information is used for generations is creating a bottleneck situation. This is because one particular set of genetic information is being continually propagated and the diversity of one spore-match's genetics are limited.
This would be circumvented by using a print from the cloned mushroom - where you would no longer be in a bottleneck situation due to the genetic diversity contained in the various spore. Thus returning to spore-match competition.


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Re: Getting a strain isolate without agar ?? [Re: Zen Peddler]
    #1312704 -

Awsome topic.. this is by far the best thread I have read in a long long time.

So for isolation, taking a section of mycelium or cloning would be done,
to keep diversity, a print must be taken, and
to increase diversity you could mix several prints from several mushrooms.

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Re: Getting a strain isolate without agar ?? [Re: ExtravagantDream]
    #1312939 -

i am far from being a mycologist and so like some of the analogies being used here
Joshua said
"A bottleneck event is an event in which the genetics of a population is reduced greatly. Imagine 100 people are left on the Earth. Only the genes from these 100 people will be present in future generations. If all of these people were
Eskimos, the future progeny would appear as Eskimos. You would lose all other races."
then it was said
"A complete semen sample could contain your complete genetic makeup, but each cell would still only contain 1/2. A semen sample will never contain more genetic variety then your DNA because you cannot produce additional genes. This is why inbreeding leads to deformed children, the lack of genetic variation.I hope that was clear."

if there is only 100 eskimos then their children will be eskimos. so if i grow PFs there "children" should be PFs. if the eskimo children's parents try to vary every time will there be more genetic diversity? if yes, then if i grow 100 PF mushrooms and take prints is there any advantage to mixing up all 100 prints and starting multispores with that. is a spore print like a semen sample? so if i get one print and grow 100 shrooms and mix the spores is it the same as inbreeding the shrooms since the spores all came from the same "father"? is there any advantage to crossing strains, like mixing PF spores with GTs, or better still mixing all the different available cubensis strains so there is a better chance of the spores mixing. is growing a single strain a "bottleneck event" as there are no "other races" there to lose to begin with? is it better to start spores out on substrate that you plan to grow them all on. if i clone an eskimo who thrives on fish and then change his diet he may get unhealthy. i have asked this further back but got no replies, if planning on growing on rye would with liquid innoculant would it be better to start your spores in a liquid broth of rye flour since the spores that prefer rye should dominate the solution?
i hope i havent offended any eskimos :smile:

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Re: Getting a strain isolate without agar ?? [Re: blackout]
    #1313343 -

Would this not take centuries before it becomes a problem??

Isolate or not, as long as it produces better/more even/heavier flushes, i still don't see it being a problem.

How many generations of clones would it take to totally weird out a strain???

Have any of you witnessed it before??

I've heard of albino shrooms etc.... how frequent is this?

cg


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************************************.>>>>>
Here Johny,, have a joint.. all your friends are doing it!!

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Re: Getting a strain isolate without agar ?? [Re: blackout]
    #1313883 -

As Anno pointed out to me it is only the original print from a wild specimen that produces a true bottleneck event. Any isolation from those spores would only reduce the genetic variability slightly from the wild print.

I think this is what you were trying to point out to me Anno.

__________________________________________

Blackout-

if the eskimo children's parents try to vary every time will there be more genetic diversity?

Yes. This actually occurs in human population. Humans release pheremones (airborn chemical signals) that are perceived by the human body. You are more likely to be attracted to a member of the opposite sex who emits pheremones different than yours. Pheremones are a genetic trait that will vary less between people of close getetic relation.

if yes, then if i grow 100 PF mushrooms and take prints is there any advantage to mixing up all 100 prints and starting multispores with that.

No. It would be like mixing 100 of your sperm samples together v. one of your sperm samples. The sample came from the same source therefor, either sample contains the same genetic variability.

What would be beneficial to maintain genetic diversity in your shroom population is to periodically mix in wild prints from the same strain.

Agriculturists use this principle to genetically modify wheat crops. Many wheat strains have been cultured for so long they may lose the advantage of genetic diversity. A specific wheat crop may be unable to fight off a certain bug because it lacks the gene that makes the wheat toxic to the bug. There may be a wild strain of wheat that has this gene for toxicity to the bug. Geneticists will cross the domestic strain w/ the wild strain in hopes of obtaining a hybrid strain that has the favorable characteristics of the domestic strain and the immunity to the bug from the wild strain.

so if i get one print and grow 100 shrooms and mix the spores is it the same as inbreeding the shrooms since the spores all came from the same "father"?

Yes. Certain species are more sensitive to inbreeding than others. It may take P. cubensis 1000 inbreed matings before noticeable mutations occur, whereas humans can show noticeable mutatations after 1-2 inbreedings. The royal family in Russia had/has hemophilia due to inbreeding. They wanted to keep their royal blood pure and only wed to other Royal families. This led to the high prevolence of hemophilia w/i the Russian Royal family.

is there any advantage to crossing strains, like mixing PF spores with GTs, or better still mixing all the different available cubensis strains so there is a better chance of the spores mixing.

It depends on how much work you put into it. I believe Stamets says that about 1 in 1000 Oyster hybrids are more vigorous than their parental strains. These hybrids are direct matings of different strains. Mixing spores would rely on random matings which would mean you would need to do a lot of testing to find that one hybrid strain in one-thousand that is a better strain.

is growing a single strain a "bottleneck event" as there are no "other races" there to lose to begin with?

Yes. As soon as you take that first wild print, you have removed a significant ammount of genetic variation available for future matings. See my first statement of this post.

is it better to start spores out on substrate that you plan to grow them all on. if i clone an eskimo who thrives on fish and then change his diet he may get unhealthy. i have asked this further back but got no replies, if planning on growing on rye would with liquid innoculant would it be better to start your spores in a liquid broth of rye flour since the spores that prefer rye should dominate the solution?

Maybe. The idea is good. However the enzymes used to digest the nutrients that the mushrooms use will work on rye, rice, straw....etc. I think your method would help, but so little it would be hard for you to notice. It is not good to grow the same strain out on the same substrate very many times. The mycelium may not be able to get all of the nutrients it needs from a single substrate.


___________________________________________

I am a student w/ limited knowledge regarding these topics. Please take my information as such.

Joshua


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The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1313981 -

good info. But what is the solution? Say you had a single print, domestic.. incoculated several jars, and birthed each one seperatly. Due to probability, most will be different from each other and all should be different from their parent. Then mixing several prints from several different casings/cakes would increase the probability of diversity, Right? And since your only taking prints from the best of each casing/cake you are generally taking better traits (but also increasing the chance that they are the same as from the other casing). But essentially doing this over time would better the traits while remaining diverse. Or would this only maintain the diversity not increase it?

Also, If you are selecting prints from the best, why is it so helpfull to isolate? Just higher probability of creating a quality mushroom? Doesn't make much sense though. Doesn't a cloned mushroom have the same spores as the one it was cloned from?

Sorry for all the questions, it's just a really interesting topic, but rather confusing.

Edited by ExtravagantDream (02/17/03 10:40 AM)

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Re: Getting a strain isolate without agar ?? [Re: ExtravagantDream]
    #1314038 -

Post deleted by administrator.

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Re: Getting a strain isolate without agar ?? [Re: Anonymous]
    #1314060 -

How long would it take an isolate to senescing?


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Re: Getting a strain isolate without agar ?? [Re: Zen Peddler]
    #1314142 -

Post deleted by administrator.

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Re: Getting a strain isolate without agar ?? [Re: Anno]
    #1314297 -

Both prints won't be identical. They will both be random assortments of the genes of both donor nuclei from the original Dikaryon.

Both will be equally diverse, but equally limited as well. Limited in the sense that they are spores coming from the same ORIGINAL DIKARYON.

Diversity is Combining spores from as MANY DIKARYONS as posible. All of which is IRRELEVANT, because the RACES in cirulation are at best the product of 10 individuals removed from a population. How many ORIGINAL WILD PRINTS are the donors for all the STRAINS in cirulation? How many EQ prints were originally used, how many GT?

If all the printsfor a given Strain in circulation are the product of the random assortment of a single Dikaryon in the wild, DIVERSITY is non-existent, compared with the WILD STRAIN(RACE), which itself is the product of a limited number of individuals originating from another STRAIN, which is the product of another group of limited individuals originating from another STRAIN. All Strains are the product of POSSIBLY a single random mutation that created a SINGLE individual, that was from a Limited population of a STRAIN of another SPECIES.

Random assortment maintains alittle diversity, but nothing compared to the interaction of haploid myceilum within a population of MANY individuals. I.E spores of one dikaryon potentially mating with spores of 100,000 different dikaryons.

It is always better to Be picking from a population of 100 individuals, then a population of 1.

Expanded. It is better to be picking a single dikaryon formed from a population of 100 different dikaryons, then picking a single dikaryon from a population originating from 1 dikaryon.

100 prints from 100 dikaryons of a single strain, mixed into a syringe is more diverse then taking a 100 prints from a single Dikaryon from a single strain and mixing those into a syringe. Both are VERY DIVERSE. But the former has more DIVERSITY because it originated from more then a single INDIVIDUAL within a Strain.

Strain EQ from vendor 1, EQ from vendor 2, EQ from vendor 3 a print each all mixed together in a syringe is more diverse then just mixing 3 prints of Vendor1 EQ into a syringe.


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Re: Getting a strain isolate without agar ?? [Re: ]
    #1314360 -

ANNO is correct.

A print from a clone is as diverse as a print from a Multispore Dikaryon originating from the same Print. THEY are the same.

Diversity is only increased when the multispore is made up of Spores from MANY DIKARYONS.

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Re: Getting a strain isolate without agar ?? [Re: ]
    #1314594 -

>Diversity is only increased when the multispore is made up of Spores from MANY DIKARYONS.


So this would translate to:
Diversity is only increased when the multispore inoculation is made with several prints at once and the prints itself being from different multispore cultivations(=not from the same clone)


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Getting a strain isolate without agar ?? [Re: Anno]
    #1314915 -

thats what I think.. but if in theory they all came from one wild print.. can they get any more diverse?

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Re: Getting a strain isolate without agar ?? [Re: ExtravagantDream]
    #1315893 -

Joshua- thanks for the great reply. you say
"What would be beneficial to maintain genetic diversity in your shroom population is to periodically mix in wild prints from the same strain."
how would you ensure your wild prints are the same "strain", does it mean picked from the same field? as there are a few Thailand strains available but they are considered totally different strains (or are they?).

Teonan went on to say
"Strain EQ from vendor 1, EQ from vendor 2, EQ from vendor 3 a print each all mixed together in a syringe is more diverse then just mixing 3 prints of Vendor1 EQ into a syringe."
i presumed vendor A got the original print, grew it out and the other vendors bought prints from vendor A and grew them and sell them. this then contradicts this statement

"if i grow 100 PF mushrooms and take prints is there any advantage to mixing up all 100 prints and starting multispores with that.
No. It would be like mixing 100 of your sperm samples together v. one of your sperm samples. The sample came from the same source therefor, either sample contains the same genetic variability."

or should i have suggested growing 100PF mushrooms from the same syringe but on 3 different substrates, all multispore, and then mixing these to get more diverse genetics. if so are there any vendors who actively do this? would it have prevented the degeneration that the PF race suffered a while back after which it was said that PF went back to spores from years back to start decent spores again.

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Re: Getting a strain isolate without agar ?? [Re: blackout]
    #1316017 -

how would you ensure your wild prints are the same "strain", does it mean picked from the same field? as there are a few Thailand strains available but they are considered totally different strains (or are they?).

I was implying to pick the wild prints from the same area. This will secure a higher liklihood that the spores will mate. I don't want to damage any pre-existing love for the Thai strains. I am not familiar with the geography of Thailand. I would assume the strains are very similar unless the regions they were picked at are separated by geographic features that prevent the mixing of spores.

presumed vendor A got the original print, grew it out and the other vendors bought prints from vendor A and grew them and sell them. this then contradicts this statement

Yes, you are right. It is assumed the original prints that the vendors are growing from were from different wild specimens. I think it is more likely that a large percentage of the strains are from one common wild specimen. I think MJ takes several prints of specimens when he takes his trips. I would imagine there is a higher likelihood that the strains he brings back and offers to vendors are from seperate wild prints.

or should i have suggested growing 100PF mushrooms from the same syringe but on 3 different substrates, all multispore, and then mixing these to get more diverse genetics. if so are there any vendors who actively do this?

No, the variation generated by your method would be next to nothing in comparison to the parental strain. I imagine most vendors do cycle the substrate they choose to grow on...at least if they are true mycologists they would. I bet PF experienced the degradation of their strain due to very many generations being grown and on the same substrate. He could have extended the generations before mutations occured only slightly by varying his substrate. I am more likely to think that he grew the spores out so many times that he lost his variability.

____________________________________________

All of this thinking has renewed my interest in crossing some strains. Keep your eyes open.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: Anno]
    #1316880 -

I think ONLY may be a bad word to use.

Mixing ten prints from the same clone is more diverse then using one print from that clone.

Mixing ten prints, each from a different clone(Dikaryon) is MORE diverse then mixing ten from the same clone.

Mixing ten prints from ten different Dikaryons, from ten different cultivators all using different substrates is even more diverse.

Cubensis is heterothallic so it always OUTCROSSES.

For there to be a GT it must only mate with other GT spores. TheDiversity of GT is maintained by mixing spores from as many different GT prints as you can find. Some from each vendor, some from each cultivator, some from different clones from each cultivator. The more spores you can mix from one clone, several clones, or 100's of different Dikaryons(clones).


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Re: Getting a strain isolate without agar ?? [Re: ]
    #1317841 -

This of course bring upon the issue of how many prints are cruising around that are misslabeled.
So If I mix GT prints from several sources, I might as well end up growing a GTized Ecuador.....


--------------------
Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Getting a strain isolate without agar ?? [Re: Anno]
    #1317881 -

Is that really a totally bad thing? yes, if you are trying to maintane purity and looking for a true GT, or what not. But we all evolve, I'm willing to bet that GT and alot of other strains are just a mixture from some others.

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Re: Getting a strain isolate without agar ?? [Re: ExtravagantDream]
    #1317883 -

Probably. At least after some time being in circulation.

Is it a bad thing for the cultivator? No, don?t think so.


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Getting a strain isolate without agar ?? [Re: Anno]
    #1318103 -

i often wondered if 2 strains were actually the same. some vendors get "wild" prints sent to them. whos to say they were collected in the wild and not just a print from a commercial syringe? only trust i suppose.
is there any way of confirming that 2 mushrooms are different strains

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Re: Getting a strain isolate without agar ?? [Re: blackout]
    #1318166 -

>is there any way of confirming that 2 mushrooms are different strains

Of the same species?
Hardly. It?s hard already holding the different species apart.


--------------------
Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Getting a strain isolate without agar ?? [Re: ]
    #1318593 -

"For there to be a GT it must only mate with other GT spores."

For there to be an Eskimo it must mate w/ other Eskimos is not true. The heterothalic nature of GT does not require that GT cross w/ GT, only that it mates w/ a P. cubensis.

Is this right?

Also when you speak of diversity it is important to know to what extent you have diversity.

A couple allele differences in cheetahs may be considered diverse whereas a couple allele differences in humans is not. It is also important to know from where the parent dikaryon originated from. Not all dikaryon matings are of equal diversity.

I have P1A and P1B as parental dikaryons originating from P0. I fruit P1A and obtain F1A and F1B. I fruit F1A and obtain F2A and F2B. The mating of spores from P1A and P1B is more diverse than the mating of F2A and F2B.

Both matings are dikaryon matings but are not equal in their genetic diversity.

Is this right?

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1318758 -

Joshua is speaking in codes.....

If you don?t know what this is about, visit this link:

http://www.fungifun.org/sex.htm


--------------------
Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1320532 -

GT is an inbreed LINE.  To maintain it you must continue to inbreed it.  To prevent it from LOSSING ALL of it's variability you want to be able to do self-crosses with as many GT sources as possible.  YES , you have to be able to recognize GT from EQ to be able to Maintain GT.

I never stated that GT CAN ONLY MATE WITH GT, just that it must to maintain being a GT.  SELF CROSS v.s OUT CROSS. 

Of course you have to know what GT is and What EQ is.  When breeding you should know Pedigree.  This is very difficult in the OMC.  Lots of vendors, lots of trades!!!

I have two different GT's.  :smirk:






 

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Re: Getting a strain isolate without agar ?? [Re: ]
    #1320580 -

Very good then!

I know who to talk to when I want to set my understanding of genetics right.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #1320585 -

BTW - most of the various 'strains' of cubensis from Thailand are infact picked at various locations within around 10kms of one another. Diversity?


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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #2209218 -

Bump....anyone know of some cool mushroom genetics articles I could read?

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #2213086 -

jssmthrfcknchrst:

I have a comment from your quote below...

+++++
"A semen sample will never contain more genetic variety then your DNA because you cannot produce additional genes. This is why inbreeding leads to deformed children, the lack of genetic variation.
I hope that was clear."
+++++

It sounds like you are saying deformity is equal to a lack of genetic variation, but this not the case. Lack of genetic variation in and of itself will not necessarily lead to deformity. It can lead to it if harmful alleles are kept in the breeding line, but in the absence of harmful alleles the main problem with inbreeding is that a lack of genetic diversity limits the line's ability to adapt. It has a limited "tool kit" of alleles for evolution to play with, so to speak. So in the overall line, then, it might have no resistance to certain diseases, pests, etc.

On another note, there is something that I wonder about when we talk about mixing spores from various spore prints. Say someone does mix one spore print with another, and then takes a print from a resulting fruit. There really isn't any way to know that the fruit body is actually a result of spores from the separate prints mating. It could be the result of matings solely in one print or the other. Since making sure that certain spores mate with certain spores is out of the realm of most home cultivators I'm not sure what a viable option would be for most people.


--------------------
I said "no" to drugs but drugs just aint takin no for an answer!

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Re: Getting a strain isolate without agar ?? [Re: AlienPrimate]
    #2215814 -

Both deformities and the lack of disease resistance are one of the same type when spoken of in this manner. Deformities are an observable phenotype and the lack of disease resistance is a non-phenotypable genotype, both of which can be caused by inbreeding.

This is not to say all deformities or all lack of disease resistance is due to inbreeding, but both can be caused by inbreeding.

Read this link: http://www.dogbreedinfo.com/inbreeding.htm for more info.

On the second portion of your post...you are absolutely correct. The only way to be sure of a strain cross would be genetic mapping of the parent strains and of the possible hybrid offspring. One could however determine a cross, with high certainty, by observing strong phenotype characteristics under strict growth parameters. One could select two strains, one that has a blue veil and one that has a green nipple. If you crossed these strains and produced a single specimen with both characteristics, you could be pretty certain the progeny is a hybrid of the two parental strains.

Of course the above example is exaggerated to show the idea.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: Joshua]
    #2218784 -

Hi Joshua,

My comment about inbreeding was really just to highlight a fine point. When you say inbreeding "causes" deformity I assume you mean because genes that would cause it are allowed to become more prominent in an inbred line (by chance), rather than inbreeding in and of itself somehow triggering deformity. Do I understand you correctly? If not, then I am not sure what you mean by the word "causes." Unless mushroom genetics have a special quality I'm not aware of, inbreeding will only lead to deformity or what have you if harmful alleles are kept in the line through the practice. If the harmful alleles are not there to begin with, then deformity (or whatever) won't be an issue, although other problems may arise due to a lack of genetic diversity?

To I understand you correctly?

AP


--------------------
I said "no" to drugs but drugs just aint takin no for an answer!

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Re: Getting a strain isolate without agar ?? [Re: AlienPrimate]
    #2220443 -

I am talking mammals here in response to the quote you made,
"+++++
"A semen sample will never contain more genetic variety then your DNA because you cannot produce additional genes. This is why inbreeding leads to deformed children, the lack of genetic variation.
I hope that was clear."
+++++"

Fungi, I am sure, are a bit different. That is why I asked anyone if they knew of any fungi genetic articles, so I could read up and understand fungi genetics better.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Getting a strain isolate without agar ?? [Re: AlienPrimate]
    #2240073 -

According to the available literature I could find on mushroom breeding. Most deformities(lethal alleles) are worked out by the second generation of a hybrid.

IE. Breeding GT with EQ. The spores that originate from the hybrid dikaryon will contain many BAD combinations and result in a very low success rate for those spores mating to become dikaryons. The spores that su8ccessfully mate will produce fertile dikaryons that produce spores that will be FAR more viable then that original hybrid generations spores. Each future generation will get better, until you arrive back at the 1/4 succesful mating population.

When you cross GT with EQ, and we assume they do not share any immediate ancestor, the resulting spores would mate at 100%, but there succes rate will be less then the usual 25% of a SELF cross due to lethal combinations. The number will increase in the next generations until it reaches the 25% mark. Then you have a NEW strain.

Inbreeding is required to maintain a STRAIN.

Strains are themselves, inbreed populations, wild or domesticated. Cubensis will not SELF, so even if the two spores that mate came from the same mushroom, only disimilar haploids can mate to form a dikaryon. The breeding strategy prevents any SEVERE inbreeding problems. It maintains enough diversity for the STRAIN to remain viable by the number of spores produced, and the number of dikaryons resulting from those spores mating.

If the monokaryons that mate are from the same strain, they will mate succesfully 25% of the time. If the monokaryons are from different strains, that do not share a common ancestor, they will mate at 100%, but less then 25% will be succesful mates!!!

If EQ and GT are related, share a common ancestor, then there monokaryons will mate 75% of the time.

Testing 10 monokaryons of each strain against 10 monokaryons of each other Strain in circulation will determine how related ALL the STRAINs are. It will determine if they share a single parent, both parents, or neither parent.

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Re: Getting a strain isolate without agar ?? [Re: ]
    #2240091 -

Once you have established pedigree, you can continue on with BREEDING. You can find out which monokaryons of each strain do what, when mated with other monokaryons of the same strain, and also other monokaryons of other strains.

Specific traits can be breed from one strain into another strain.

The statment that hybrid strains are less then either parent is not important. The object of any GOOD breeding program is to put GOOD TRAITS into GOOD STRAINs that don't already posses the trait, and to be able to maintain the trait.

Some Hybrids might not pass the trait on to there offspring and may only be able to maintain that trait via cloning of the original hybrid Dikaryon.

I think the most important thing is to determine pedigree. It would be nice to know how related each STRAIN is, and we might be able to determine which Strain is the original ancestor of all the strains in circulation. There might be an EVE amongst the strains in circulation. A single strain that can only mate 75% of the time with all the other strains. Then again there might be some strains that have lost all ability to mate with other strains, ie multiple species emerging from a single species. Psilocybe subcubensis, Psilocybe cubensis, Psilocybe ?

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Re: Getting a strain isolate without agar ?? [Re: ]
    #2240453 -

I love genetics!!!

Thanks for the info. Do you know of any good reads when it comes to fungi genetics?

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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