BorisTBD said:

Beginner here,

A bit overwhelmed with this agar work. Not sure what to do next other than more transfers of course but I am now sitting on 30 plus plates that look very similar to the first three pics. First three are 1st transfers. Last two are syringe.The first pic looks great! Should I make any transfers from it? The second pic has the fuzzy undeveloped part at the 1 to 5 region. Is it “Monokaryotic, or homokaryotic mycelium”?(I read that some where) Or is it mold? Pic three has the same fluffy stuff. I’ll be making transfers from the 3 separate regions at 4 to 9 and also 10 to 1. I have already made transfers from other similar plates and day 3 shows same pattern of growth. So far I have only one GOOD symmetrical plate. Do they all need to be like the plate in picture 1 before moving to grain? Last two pics show what seems to be mycelium on top of bacteria. After a few days of the syringe drop my plates had milky wet spots. Then a few days later white fluff popped up on top of it. I was going to make transfers from pic 4 but after looking at the bottom it’s all black now (pic 6)and may b too late. Should I even attempt a transfer when knowing for sure it’s contaminated underneath? Thanks again for any help.

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You could make a couple of masterjars with the first plate.

Id take a couple of wedges from the second to expand and if they are the same culture try to lose as many rubbish plates as you can.  Yes, you can leave a few to fruit but if you get it right you should have fruits from a grow in a month/6 weeks where u can choose you fruit you want to clone. 

Saying that look at this bad boy from a bacterial jar.  Couldnt resist cloning him!!!!

spiritlands said:

Boris, I would just use several pieces from the first plate and carry on with however you plan to grow, like lc, grain, slurry etc. Id set the next best one aside and just wait till it tries to fruit and I clone that for later. I start several dishes at once but I only use the very best, it's ok to not take transfers from everything. I know what you mean about the wet spot from the syringe plate. I think sometimes it just looks like that at first due to the excess water and the Mycelium starting to grow. Sometimes it is bacteria. If it is bacteria you can pour hot peroxide agar over top and the Mycelium will recover and poke up through the top, you can then transfer again and clean it up. This is only for when you must clean dirty tissue. Try to just be sterile every time.
As for your question on monokaryotic/ dikaryotic... Statistically all Mycelium you work with will be dikaryotic. The only way you would have monokaryotic is if you isolated a single spore and germinated it. Any time two monokaryotic Mycelium strands meet they share genes and become dikaryotic.

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BorisTBD said:

I plan on using grain. So, it would be safe to assume that these plates are clean and ready for grain? Shouldn’t the first plate grow out a little more? Is it possible to cut it into 4 pieces for 4 rye jars?

- I would use the first plate and cut it in half.  6-to 12 looks the best.  Get a clean jar and then g2g

How should I fruit the Petri dishes when they are fully colonized? Remove the lid and place upright in a ziplock bag then mist the sides? Sort of like a mushroom grow bag?

-Just leave it and it will fruit itself

I’m still slightly confused about dikaryotic. So is the white puffy part monokaryotic then mates and becomes the vein looking dikaryotic aka Rhizomorphic?

-monokaryotic is when u have a mono culture, its like 1 rule for that mycellium.  If that rule was fat and tall all the fruits would be fat and tall.  Forget all this stuff at the moment just try to get clone cultures and fruit them.  Mono cultures take a long time to get just to test to find out they are useless.  Clone culture succeed lots of times.

Must I wait for it to be rhizomorphic before transfering or will the puffy part eventually turn into the vein looking rhizomorphic stuff?

-The stuff on plate one is rhizomorphic

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Edmunter said:

BorisTBD said:

I plan on using grain. So, it would be safe to assume that these plates are clean and ready for grain? Shouldn’t the first plate grow out a little more? Is it possible to cut it into 4 pieces for 4 rye jars?

- I would use the first plate and cut it in half.  6-to 12 looks the best.  Get a clean jar and then g2g

How should I fruit the Petri dishes when they are fully colonized? Remove the lid and place upright in a ziplock bag then mist the sides? Sort of like a mushroom grow bag?

-Just leave it and it will fruit itself

I’m still slightly confused about dikaryotic. So is the white puffy part monokaryotic then mates and becomes the vein looking dikaryotic aka Rhizomorphic?

-monokaryotic is when u have a mono culture, its like 1 rule for that mycellium.  If that rule was fat and tall all the fruits would be fat and tall.  Forget all this stuff at the moment just try to get clone cultures and fruit them.  Mono cultures take a long time to get just to test to find out they are useless.  Clone culture succeed lots of times.

Must I wait for it to be rhizomorphic before transfering or will the puffy part eventually turn into the vein looking rhizomorphic stuff?

-The stuff on plate one is rhizomorphic

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BorisTBD said:

Edmunter said:

BorisTBD said:

I plan on using grain. So, it would be safe to assume that these plates are clean and ready for grain? Shouldn’t the first plate grow out a little more? Is it possible to cut it into 4 pieces for 4 rye jars?

- I would use the first plate and cut it in half.  6-to 12 looks the best.  Get a clean jar and then g2g

How should I fruit the Petri dishes when they are fully colonized? Remove the lid and place upright in a ziplock bag then mist the sides? Sort of like a mushroom grow bag?

-Just leave it and it will fruit itself

I’m still slightly confused about dikaryotic. So is the white puffy part monokaryotic then mates and becomes the vein looking dikaryotic aka Rhizomorphic?

-monokaryotic is when u have a mono culture, its like 1 rule for that mycellium.  If that rule was fat and tall all the fruits would be fat and tall.  Forget all this stuff at the moment just try to get clone cultures and fruit them.  Mono cultures take a long time to get just to test to find out they are useless.  Clone culture succeed lots of times.

Must I wait for it to be rhizomorphic before transfering or will the puffy part eventually turn into the vein looking rhizomorphic stuff?

-The stuff on plate one is rhizomorphic

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I’m excited! A2G tonight then my good sir! And for the potential pinning Petri dish, is it considered fully colonized when the agar is completely taken over or just the top portion?

- when the entire top is white

-Just leave it and it will fruit itself
Do I remove the Petri dish lid once it starts to show signs of pinning?

-Dont touch them and one day you will see pins.  Some grow a fair size.  The genetics are usually banging from fruited mushrooms on plates.  Dont open them and transfer clone material them to clean plates.

I’m probably missing something as usual but please spoon feed me this last question. What should I call that puffy undeveloped white part?  I

-its either contaminated or tomentose mycellium

Thanx for the help!

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BorisTBD said:

How should I fruit the Petri dishes when they are fully colonized? Remove the lid and place upright in a ziplock bag then mist the sides? Sort of like a mushroom grow bag?

I’m still slightly confused about dikaryotic. So is the white puffy part monokaryotic then mates and becomes the vein looking dikaryotic aka Rhizomorphic? Must I wait for it to be rhizomorphic before transfering or will the puffy part eventually turn into the vein looking rhizomorphic stuff?

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Edmunter said:

BorisTBD said:

-monokaryotic is when u have a mono culture, its like 1 rule for that mycellium.  If that rule was fat and tall all the fruits would be fat and tall.  Forget all this stuff at the moment just try to get clone cultures and fruit them.  Mono cultures take a long time to get just to test to find out they are useless.  Clone culture succeed lots of times.

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spiritlands said:

Edmunter said:

BorisTBD said:

-monokaryotic is when u have a mono culture, its like 1 rule for that mycellium.  If that rule was fat and tall all the fruits would be fat and tall.  Forget all this stuff at the moment just try to get clone cultures and fruit them.  Mono cultures take a long time to get just to test to find out they are useless.  Clone culture succeed lots of times.

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Totally agree with the last statement to not worry about it. But a mono culture is an isolate, a clone. And everything you said there is correct except that an isolate or clone from a selected fruit is not monokaryotic, if it were it would be unable to fruit. All fruits are dikaryotic. The word mono is in it but they're unrelated.

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LtLurker said:

Clearing up terms. . .

A clone is just a clone, it can contain 1000's of strains.

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Can you point me to any microscopic research that confirms this? I have been led to believe this is not the case and apart from the growers who make assumptions at a macro level, this seems far from intuitive.

Each of the 1000’s of strains would have to be growing at almost exactly the same speed to be maintained after multiple leading edge transfers, which seems particularly unlikely considerring they are all competing for resource at the same time. wouldn’t he who gets the best resource start, take the lead, taking the lions share of the nutrient as progressing, promoting stronger growth?