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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Innoculating a jar with multiple strains
    #571909 -

In the FAQ about innoculating a jar with multiple strains it states that doing so would result in the domination of one strain over the other or a cake fruiting both types of strain. I thought that sexual reproduction of a fungus occurred when mycelium of two "individuals" met and exchanged genes. Is it possible for this to happen and "cross" two strains? Just curious.

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Re: Innoculating a jar with multiple strains [Re: zanchin]
    #571989 -

They produce sexually as the spores of the same strain mate and produce the mycelium... Like the FAQ said, you'd either get the domination of a strain, or they could stick to their own sides and see both. They wouldnt just cross without cloning them in agar.

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Re: Innoculating a jar with multiple strains [Re: zanchin]
    #572015 -

When multisporing with a single Strain, the resulting substrate has many sub strains growing in it. Some of these substrains remain there own, some fuse to become new substrains, and some just get outcompeted.

When combining spores from two strains, and multispore innoculating. The exact same thing happens as mentioned above, within each strain, and possibly between each seperate strain. Hybridization is possible. The problem is, you would never know it has occured!!! Hybridization is more likely to come from Anastomosis, fusion of Dikaryons, within a single strain, and then maybe between individual Dikaryons of the two seperate strains.

Spores clump together, and germinate together. They mate with the closest compatible monokaryon they come in contact with. So the likelyhood of getting a germinated spore from one strain to mate with a germinated spore of another strain is highly unlikely with concentrated multispore innoculations.

Anastomosis is going to happen for sure within the single strains dikaryons, and will probably occur with lesser frequency between the seperate strains.

Individual Dikaryons compete for substrate. The ones that are best suited for using the environment they are placed in, win the competition. Hybrids tend to be less vigorous, so even if you get one to occur between the two distinct STRAINS, it will probably be out competed by the remaining Dikaryons. The dominant Dikaryons will win the race. Dominance in the sense of Vegetative growth. There is no way of knowing if the winner is the best for fruiting, without testing it against other Dikaryons, within the same environmental parameters.

If you are interested in breeding, AGAR culture is a must. Spore dilution is EXTREMELY helpful. Lots of time, and money will certainly help as well.

You can either stick with a single Strain, and do controlled matings of monokaryons. Testing all resulting Dikaryons for fruiting ability in your environmental parameters. Or you can test all the monokaryons for compatibility with monokaryons from another Strain. Testing fruiting ability of all suspected matings.

Another option is to put a single isolated dikaryon from one Strain on a plate with a single isolated dikaryon from another strain, testing for compatibility and anastomosis. You will have to test alot of isolates!!!

The MAJORITY of HYBRIDS are LESS VIGOROUS then their donor STRAINS. ROLL THE DICE.

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Re: Innoculating a jar with multiple strains [Re: ]
    #572026 -

Germinated spores of cubensis are typically uni nucleate. They are monokaryons. They grow out until they come in contact with a compatible monokaryon to mate with. There is a fusion between these monokaryons, but genetic information is not exchanged at a nuclear level. The two contributing nuclei exist under a single bi nucleate hyphae, seperately. The only time these nuclei fuse is in the basidium of a mature fruit. This is the sexual reproduction. The two nuclei fuse then undergo meiosis. The resulting spores are reduced to the haploid state.

These germinate and the process repeats itself.

When a monokaryon from one Strain mates with a monokaryon from another strain, there is still no exchange of genetic information at a nuclear level. The haploid nuclei from each donor strain is incorporated into a binucleate, dikaryotic hyphal strand. The two nuclei remain seperate!!!! They only fuse in the basidium of the mature fruit.

Problems: Monokaryons that are not completely compatible fuse. These do NOT result in fruiting DIKARYONS.

The nuclei are haploid, They contain only half of the genes necessary for fruiting. The two nuclei exist seperately, and work independently of each other. It is there combined effects that make it a dikaryon.

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Re: Innoculating a jar with multiple strains [Re: ]
    #572042 -

If your interest is primarily in finding a good substrain of a single strain. Do a bunch of multispore innoculations, via syringe, into a bunch of jars. Clone the best fruit from each jar, and then test all of these against eachother, seperately, under the environment you will be growing them under. This will solve all of the PROBLEMS associated with false matings between partially compatible, but non fruiting, matings!!!

If your interested in breeding, I suggest doing alot of reading to help define all the terms that have been mentioned above!!!

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Re: Innoculating a jar with multiple strains [Re: ]
    #572112 -

so, with the right equipment, it is possible to breed spores together of different strains? Could you breed azures with cubensis, or would that not work because the species is different? If it is possible maybe I could minor in mycology and truly bring the PESA to life.


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"...Three geese in a flock. One flew east, And one flew west, And one flew over the cuckoo's nest."
Ken Kesey

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Re: Innoculating a jar with multiple strains [Re: angryshroom]
    #572185 -

Why cloning? Why Agar?? I have had great success with cloning without using agar.
Teonan - I have a friend who is doing a PHd in biology - would he be right in saying that when you clone from one fruit in a multispore colonised substrate, you get a clone of the entire individual from its spore-match - which could have the characteristics of the one fruit - or the characteristics of them all - in that the individual could be responsible for one fruit or all of them??


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Re: Innoculating a jar with multiple strains [Re: Zen Peddler]
    #572465 -

Hybrids between species? I don't think so as of yet!!!

Sure you can clone without agar, but you will have trouble isolating monokaryons without some sort of two dimensional surface to germinate spores on.

There are agar substitutes as well.

When you multispore innoculate a substrate, you have many substrains growing. Some of them are completely incompatible with others, THEY WILL NOT BRANCH, or FUSE in any way shape or form. OTHERS WILL FUSE TOGETHER.

When you clone a single shroom from this colony, it can be an identical match with every shroom in the colony, produced. It can also be disimilar to all the fruits in the colony. You JUST ARE NOT CERTAIN WHAT HAS HAPPENED. Within a single SPORE RACE, the variability of mushroom appearance grown on the same substrate in the same environment is going to be negligible. So it really didn't matter which mating one the race, or which matings fused together, or which matings were totally incompatible, and got outcompeted.

All clones do, is guarentee the purity of the strain. It is a single Dikaryons fruit. What occured in the multispore matings to get the final dikaryon that produced the fruit, is in all essence not important.

Just pick several fruits from the single tray, and several fruits from all the other multispore trays, or cakes. Clone them, any way you like!!! Then innoculate new jars, with one strain in a jar, and Test them against each other.

The variability you are looking for, is not the mushrooms appearance, but it's preformance in the conditions you grow in. This is the variability that exists within a spore race.

To answer your question, I think? A multispore innoculation can result in a single dominant strain doing all the fruiting. It can also result in many strains doing the fruiting. It can also result in entirely new strains doing the fruiting, if by chance you had two different strains growing in the same substrate, and they undergo anastomosis. Two dikaryons from different strains fusing together, and exchanging Nuclei. A1B1 from one strain going with A2B2 from the other strain, and vice versus. But individual clones come from an individual dikaryon, and they contain the genetics of only a single compatibly matched Dikaryon.

You can have several different strains fused together, but the fruitbody is going to have the characteristics of a single dikaryon, not all the dikaryons present in the colony.
As the colony approaches the fruiting stage, hyphae often contain MANY NUCLEI within the actively growing hyphae. Many more then two. But they all segregate back into the normal binucleate state prior to the formation of Basidia, and Karyogomay, meiosis, and spore production.

The short answer is Dikaryons that recognize eachother can and do interact. But Spores originate from a single Dikaryon.

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Re: Innoculating a jar with multiple strains [Re: ]
    #572478 -

Let me add that you can have freak shrooms form from a random mutation. A shroom with some morphological characteristic that seperates it from the rest of the spore race. Cloning this will result in all mushrooms carrying this trait.

Example polyploids.

Then again if the mutation is sectional, or only existing within some of the hyphae that make up the shroom, then it will not be expressed in a clone, unless the tissue cloned all has the mutation. This kind of sectional mutation might not work on it's own though.


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Re: Innoculating a jar with multiple strains [Re: ]
    #572496 -

Teonan, you have been most helpful. I think I will try to do a bit more reading on the subject as you have suggested. My ultimate goal is to try to engineer a strain or substrain that grows best at cooler temps. Maybe artificial selection. More reading before anything else though. Thanks a lot.

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