49 replies to this topic

[Microbe]

Creating a double haploid or doubling the chromosomes in a single monokaryotic culture is something i was researching but i needed a break.

That is very interesting, I would love to hear where your research led.

If successful it would open the doors wide open for selective breeding including crossing or creating hybrids by being able to truly control the lineage.

Exactly, overcoming the mating types and achieving complete homozygosity would be a game changer!

I have noticed that a dilution onto agar followed by lowering the incubation temp, makes the germination play out a lot slower and you would have a bigger window to grab cultures as they germinate.

Typically when we see mycelium growing, it is very probable that is dikaryotic or already mated. By spore dilution i mean taking a piece of nicrome wire and touching a spore print then swirl it around in 100 ml or more of water then aspirate the water into a syringe then disperse single drops via 27 gauge sharp onto agar. Couldn't imagine how many drops that would be. Just talking about it makes me anxious about it allover again. Im trying to balance life and a new career at the moment but once i balance it, i will hit the mycology with intensity again.

Edited by Microbe, 15 March 2019 - 07:30 AM.

[PistolPete13]

They also use water agar and slightly lower incubation temperatures, so you do not get such fast growth. It would be interesting to see if those $20 usb microscopes you can get on ebay that say they get 1600x magnification, would pick up a spores on a white background?

 

I am looking forward to your return, sounds excellent! 

[Phungivore]

got one them scope have break out print and see what i can see. years ago my pappy was talking about using cholchine to produce super weed plants, sounds like similar affects to fungi too. pretty cool mikey

[PistolPete13]

Hi Phungivore it would be great if you could take a few screen grabs and upload them here to show us what they can do. I noticed there were different magnifications, the best I could see and I did not really look hard was 1600X magnification which in theory should be fine. But in practice they may not work so well???

[Seeker2be]

Thank you for all your detailed  research!!!  Your ARX   

[Seeker2be]

Mikey:  You note that ARX produces few spores but I obtained 8 spore prints today with no problem  from a first flush.  The prints are red like redboy.   While the criticism of colchicene  as toxic was elaborated upon and well merited  I would think that  generation there would be  no colchicine incorporated in the fruitings? No?  Sort of like feminized seed treatment?

[jkdeth]

Thought I'd drop mine in. I was having a horrible time with agar when I started this, (Not related to the ARX). So I bit the bullet clipped my portion of print in half and made some MS.
I ended up tossing some grain gone bad, but got 1 quart jar colonized and one that contaminated at around 70 percent. Contrary to any advice anyone has ever heard read or given. I spawned what good lumps I could get out of the bad jar. I ended up with a pint of spawn divided between 3 pint size Dub tubs with coir/verm substrate. The first produced a flush of tiny mushrooms, second flush provided a cap large enough to print.

The send has a a larger decent cluster I'll clone. And a very "APE" fruit. I may clone that too. The third is still in small pin stage.

[Seeker2be]

Nice, just sent some prints to Australia, Africa and soon Spain.

[Misfit]

Wanted to contribute to the post.
Arx ms to grain all contamed. RedboyX went much better. I have 6 or 7 shoeboxes running. They were all ms to whole oat. Spawned to coir/verm. Cased in peat moss/verm/oyster shell. I took that casing from one of DonShadows more recent TEK posts. I was using his substrate recipe as well but a lot of people grumbled about coffee being a mold magnet. So I decided to keep it a bit more simple for the time being.
I will get some pics of my pinsets later after work.
So far they seem to be at least 95% leucistic. There are a few with pigment here and there. Some fruits have matured earlier than the rest of the tub so I harvested them. They don’t look like the other cubes I have grown.
I put several samples down the print last night from this first group

[Moose2141]

Creating a double haploid or doubling the chromosomes in a single monokaryotic culture is something i was researching but i needed a break.

That is very interesting, I would love to hear where your research led.

If successful it would open the doors wide open for selective breeding including crossing or creating hybrids by being able to truly control the lineage.

Exactly, overcoming the mating types and achieving complete homozygosity would be a game changer!

I have noticed that a dilution onto agar followed by lowering the incubation temp, makes the germination play out a lot slower and you would have a bigger window to grab cultures as they germinate.

Typically when we see mycelium growing, it is very probable that is dikaryotic or already mated. By spore dilution i mean taking a piece of nicrome wire and touching a spore print then swirl it around in 100 ml or more of water then aspirate the water into a syringe then disperse single drops via 27 gauge sharp onto agar. Couldn't imagine how many drops that would be. Just talking about it makes me anxious about it allover again. Im trying to balance life and a new career at the moment but once i balance it, i will hit the mycology with intensity again.

[Moose2141]

What is nicrome wire? Just a really small gauge wire? Is it a special alloy? Would a sewing needle be too fat to be effective?

[Moose2141]

Awesome thread by the way!

[Misfit]

What is nicrome wire? Just a really small gauge wire? Is it a special alloy? Would a sewing needle be too fat to be effective?

Nichrome is just the type of alloy used. It comes in different gauges. It’s usually used in foam cutters and vape coils. At least the things I know of.
Edit: you can use a paper clip if you want. You just need something to scrape and transfer the spores.

Edited by Misfit, 26 August 2019 - 08:53 AM.

[Seeker2be]

MS spore to grain is a crap shoot.  Agar is easy and avoids contamination.  Happy to help if you need agar advice.

[Microbe]

What is nicrome wire? Just a really small gauge wire? Is it a special alloy? Would a sewing needle be too fat to be effective?

Nichrome wire is made up of 80% nickle and 20% chrome. It is used in many applications where repeated heating and cooling cycles are common because it resists the damage to its structural integrity that is often associated with other alloys when heating and cooling cycles are present.

When i say wire im referring to a very thin single strand. The more surface area you have on your tool the more biomass that will accumulate when you touch it to a culture.



“It is always the simple that produces the marvelous.” —Amelia Barr

Edited by Microbe, 26 August 2019 - 12:33 PM.

[Julianie]

I have a shoebox running ARX right now, its just beginning to pin. Pics to follow in coming days. 

I went from spore to agar to SOFT winter wheat. Roughly 1:2 straight coir/nothing else. 

No agar to agar transfers for me, I dropped from the first dish into grain. 

[coorsmikey]

I have a shoebox running ARX right now, its just beginning to pin. Pics to follow in coming days. 

I went from spore to agar to SOFT winter wheat. Roughly 1:2 straight coir/nothing else. 

No agar to agar transfers for me, I dropped from the first dish into grain. 

 

Ha too cool! I hope you share some pics for reference. I especially hope you share the experience too! I'm looking forward to it.

[Julianie]

Odd looking fruits and very small. These are about size of dimes.

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[Julianie]

I'm thinking I should have done a couple agar transfers before going to grain on this 1....
So I did just that from the petris I kept in the fridge.

When I get some stronger growth I'll drop into grain again.

Edited by Julianie, 17 January 2020 - 10:41 AM.

[coorsmikey]

Is that a coir only substrate?