Breeding Mushrooms Part 2: Mating Haploid Isolates to Form Diploid Mycelium 

I get so excited seeing Uni-level stuff that I've done being actively supported and disseminated on youtube! I get so excitd i want to comment in the first 5 minutes of videos but always wait lol

Thank you, you sparked my interest and I will be experimenting with your techniques to create new genetic strains. It's crazy that you are the only person on RU-vid who talks about this amazing process. Who would have thought haploid only has one chromosome set and we have the ability to create 2 complete chromosome sets. SO MUCH FUN!

This is ART, Mr. White!

Thanks for this Gary! Super interesting and exciting stuff.

I really appreciate this thorough and methodical display of the practice, thank you!

Absolutely awesome mate. Thank you so much for putting this stuff out. Absolute quality!

This is incredible knowledge you are sharing. I've got what I need now to get started on something I've wondered about for months. Thank you!

Gary , wonderful Video, i love your organization. you make it seem very easy. THANK YOU

Damn your good bro! Your agar process may be the tidiest and quickest I’ve seen yet.

Love it. Thank you for your generosity and time

Thank you everyone now go out there and breed some cool strains! MUSHLOVE

Thank you for putting this content out here.

Keep the videos coming. Thank you.

Iv been waiting for this day

Gary, you are the man. Thanks for all the kick ass videos. I'm learning a phenomenal amount. Good karma for FFTFF!

Hey Gary - thanks so much for sharing this! I am bout to nerd out like a boss! 😜😆

OK coming from a small business owner making spawn, some one in mycology for 6 years and some some who have gone through almost everyone on RU-vid with mycology I take my hat of for you, your methods for explaining is the simplest and easiest

Thank you for sharing your knowledge! I cant wait to try this.

Awesome video!

Good information, grettings from Chile!🇨🇱

17:56 the one time you didn't get it perfect is how my transfers look everytime hahaha

Beautiful! Thank you!

We're all very excited about your chestnuts too.

This is awesome... thank you!

Great content. Thanks for the time🌈😃🤙

Hey Gary, I’ve really been enjoying these videos. You seem to be extremely well educated and a good presenter. May I ask if you know of any books or other robust resources that I could study to learn about this genetic/breeding/microbiology stuff pertaining to our mycology contexts? RU-vid is a great resource, but I really like an authoritative sources like books

Very helpful!

Thank you for sharing your knowledge!!

Great!

Do you look for clamp connections under a microscope to make sure u have monokaryotic isolates before introducing them together? Or do u not find it necessary

Will you take a transfer of the mated diploids and let that grow out or just put the diploid plate straight to grain? And what do you do w/ the plates that don't mate and remain haploid?

Nice

Great video! Quick question will Haploid and Diploid Mycelium act and look the same if grow out the only difference being one can produce fruit? Is there like a certain distance a Haploid can grow out looking for a mate before it stalls or looses viggor? to the naked eye do both behave and look the same and there is no way to tell one from the other?

I think the term is eukaryotic and dikaryotic mean single celled nucleus and double celled nucleus.

Please do a video hybridizing yellow and pink oyster mushrooms!!

Does each dish with isolates contain dikaryotic mycelium? Looks like those mycelia in dishes come from mushroom body fragments, not spores.

Please could you explain how that is haploid?

Is there an ideal amount of space to leave between transfers for them to grow into? Or could they be placed directly next to each other in hopes that they find each other and mate sooner, leaving more room for peripheral growth?

are the isolates you are working with in this video the same spore dilution plates from video one feels like I skipped a step or video

Not me dreaming of breeding mushrooms in the mountains for the rest of my life lol.

so that means mushroom throwing spores and they creating haploid colonies then finding each other and create new diploin breed ready for fruiting, right?can we say that diploid colony needed for mycilium to fruit? thank you so much! i'm really appreciate you sharing such information

this is just a random question im sure you get all the time definately in the mindset of coulda not shoulda but could you cross breed say pink oyster and a cubensis variety? would they even be able to breed? for purely scientific purposes of course

This video got me price checking pyrex petri dishes

Hey! I was wondering. Do you also do this for your Cordyceps?

Also, is there a rule of thumb in regards to what species will mix?

Do Cordyceps militaris spores last long? Can you use them to make a spore print on foil and store it for years? Or should the spores be planted immediately since their shelf life is short?

How do you know its a haploid, and two didn't mate? Is it very obvious? Does the look vary wildly across species? Thanks

Ol big brain over here flexing. Many thanks homie 🤙

Hello! Do you think this is possible to do in a still air box?

What type of agar is that? It's so pretty

Who taught you all of this? very cool

Question - that was a pretty big gap between video 1 and 2 - how did you isolate the haploid mycelium and know it was haploid? I saw you mention in another question that you either have to fruit it out or genetic sequence. Which did you do? Seems like a lot of work/waste to "fruit" stuff out to see if it doesn't fruit to make sure it's hapoid, especially when you know nothing else about the strain.

Good job! Please can you tell me souce where I can find information about making powerful mycelium with good crop. I know how to make pure culture on agar media from spor or tissue but how I can know that it's profitable strain?

How do you know if you have a haploid though what if two spores just stuck together after diluting and you didn't know? Can you tell?

instead of using all those scael blades couldn't you have used flame to sterilize 1 blade? ty

Have anything against high-temp sterilizing your blade and just using one?

OK the numbers are the order in which they formed the colony, what is the letter code?

I want to know these referrals and their amounts?

Hey, Thanks for this! quick question - How can I tell if my culture is haploid or diploid without a microscope?

This makes me wonder if a microtubulin inhibitor like oryzalin could produce polyploid fungi.

what did you seal your dishes with?

Do you do microscopy to confirm monokaryons?

I hate to nag, but could you do a lab walk through, with afterthoughts? I'm starting my basement lab and would like tips from someone that's proven successful 👍 I've gotten a reply in regards to the flow hoods too, prices were on point 😊 what width is yours?

Is this the morel mushroom production process?

I hate how life doesn't allow me to be first on these videos 😅

Is every colony from spore plate from streaked solution a haploid colony definitely?

Providing a link to the conclusion that would be good idea

What results of isolation in part 1?

It would help if you showed some results of your experiments

How can I modify the genetics of my mushroom?

what are the materials used

I think I'm finding out that the cotton looking mycelium is from too much moisture . And the (slow growth/growing more in a mound /not stretching) is because u have too much nutrients for the mycelium to eat

is diploid mycelium and dikaryotic mycelium interchangeable?

Отлично брат, ты супер!!!

you don't ever worry about contam with handling those with bare hands?

Can you ship to Canada?

Great content, but honestly I wouldn't waste so many blades. Just flae-sterilize the quickly or with ethanol.

I'm still unclear on how you made sure the haploids were truly single haploids and not 2 or more or already fused diploids. Did you just lower the concentration of spores so much that they were too dilute in your streaks to meet and fuse before you separated them?

18:55 yeah close call. I even held my breath when I saw that slip.

This is how you cross breed lots of different isolates of sepperate strains, love the lab coat and the apple earphone but you're not fooling me. Using 6 different scalpels when you could have used one and flame sterilized it each time which is what you should be doing if you hope to achieve selecting isolates from the agar...

Yeah, I'm so lost.

Bro, you really counted all the permutations? There's a whole field of math called combinatorics and you can calculate that with a simple equation: N x (N-1) / 2

Let me get some things straight, why are you assuming these are haploid isolates? As far as i can tell you just streaked out from serial dilution and the thing with mushroom spores is they stick.. so chances are each of those colonies are actually dikaryotic (not diploid like you said) unless you confirmed they were single spores through microscopy. How are you confirming these were actually single spored

God I'm really scratching my head right now . So I'm watching your videos in for some reason I'm having a hard time grasping and taking it all in maybe I'm stressed. And I've been looking up a lot of research on you know mixing mushrooms and some people say snake that some are even saying utilizing penicillin I believe. I was watching one of your videos and I'm not sure if I grasped it right but it seems like you were taking two different species putting them inside the same petri dish and if the mycelium (connected in the middle You're taking a sample from the connection) And if the two mycelium mats were not compatible clearly divide down the middle. Now with that being said let's say I wanted to crossbreed Panaeolus cyanescens with something like albino penis envy. Could I just take the two mycelium grow them out in the dish and And if it connects in the middle take a sample and put it into a different dish and then grow it out? My question is from that connection sample from ape an pan can is there a chance I could crossbreed or make a hybrid penis envy/pan . Or I might just completely wrong about why you are putting two different mycelial mats together hoping for a connection? I'm sorry I'm just trying to take this all in

Meow.

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and you should close your petri dishes between transfers

Damn....what happened to this guy's flow hood? Looks like someone kicked the livin' shit out of it. Disgruntled employee? Bad day at the office? Too much whiskey at the staff Christmas party? 6:01

biggest waste of plastic and blades... use pp5 dishes and reuse them !!!

I heard genetics can't cross that way.

dude, you really need to try and practice more sustainable less wasteful methods. I appreciate your videos, but the amount of plastic waste is just ridiculous