Selecting phenotypes and sectoring mushroom mycelium on agar (breeding mushrooms from spores)
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- Опубликовано: 3 янв 2022
- This is a very exciting video for our 2022 mushroom breeding project when we select the healthiest mycelium to grow out new phenotypes.
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5 day read on plates (peroxide experiment) • Germinated Mushroom Sp... 
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12:30 Colorado Oyster Isolate Selections (for those later viewing this over again)
Here's a tip for you if you want it cheap extra incubator well you can build them but if you don't want to go through all that hassle get a dehydrator they're like a hundred bucks on Amazon for a decent size one and put plates in a plastic bag set your temperature and let her go restart it after 24 hours cuz I think that's a maximum time on most of them but I mean it's the perfect incubator they're very accurate the temperature controllers on the most part and if you don't trust it try it with a second thermometer or some kind of thermocouple but I found they're very accurate and they work perfectly hell of a lot cheaper than a commercial incubator and they hold quite a few plates. I mean with the plastic bag it takes up a little more room but it's still well worth it for a cheap solution
Hi Gary, I'm an italian PhD students and I'm working on mushrooms breeding. Your videos are very interesting because they are very specific, useful and pratical. Thank you so much
It’s good to see I’m not the only 1 who gets some wrecked agar plates.
Holding life in your hand is something special . So much opportunity.
This channel is so comprehensive and detailed. This is an amazing resource for beginners like myself. Great job sir and thank you!
I've watched lots of your work and this by far is my favorite video!! Intense bro!! Freaking awesome 👌
Im studing books, but a video like this makes it so much clearer, thank you
Very thankful for your time you dedicate to the love of mycology.
so scary having that trich plate in your awesome lab. MUSHLOVE!
hah it was instantly disposed of - deep cleaning in progress 😂
Love your vibes and videos :) makes my day !!!
Your Channel is awesome Dude. Very valueable content - thanks for all the effort!! Learning A LOT from Ur vids. Best wishes from Stuttgart, Germany mushloooooove🍄🍀
Hi Gary been following you for a while loveing you shearing your skills great info thanks so much
Thank you for all your videos. New to this and love how you teach us
Love you gary!
I like mycology, with your knowledge, I still like better, Thanks very much Gary, you are great.
thanks for watching and following along!
My dude! Great work. Easily the most informative content I’ve found on RUclips 👌
Congrats!!!! ☮️🙏🏻🍄
Useful video as always!:)
Glad the peroxide worked out, somewhat. God bless!
Amazing information as always thank you so much!
Oh wow, i have got to learn a lot more! I have been experimenting with 'segmenting' without fully understanding what's going on. Sheesh! Thanks for this vid!
Just realized who you remind me of, Dave Foley from KitH. I'm a fan of Kids in the Hall and of your stuff so it is by no means an insult.
A huge thanks to you and other folks out there who teach people.
I see it a bit. I used to absolutely love Kith
I FRICKIN LOVE YOUR ACCOUNT THANK YOU A LOT FOR SHARING!
thanks for watching! 🍄❤️🙏🏻
Excellent clinical grade work. I hope to be able to copy this thanks.
Really appreciate the info my good sir. Gracias
good stuff bro. this isnext level
Got your culture syringe yesterday. About to start a couple projects from it. Thx man
I swear I grew cordyceps from a liquid culture syringe that was over a year old!! I used the egg substrate for 6 jars and rye for 1. Lost 1 jar out of 7 contamination. But, the rest are going strong and I'm about to take spores tonight from the first and best jar to mature. The fruits are all very short, thick and wild looking. I'm delighted they did as well as they did with a culture I almost threw away! #MUSHROOMFANTASIES
Cool techniques
Very helpful video. TY
thanks for watching!
Yo bro... you are the coolest nerd!
Thanks Gary. Once again, some great info. I know you are super busy, but if you get a minute or if others have the answer. In some cases, you sectioned out just the mycelium edges vs. what I'll call an entire body (colony?). Was that selection choice a function of Isolated vs. mated or was it just a function of size of sample?
Dear i am from pakistan learning hybread your videos are interesting explain hybrid for learners. thank you
Do you have a video on your huge flow hood? Thanks man, keep up the good work!
Now the year is over. Would love to hear your thoughts on the breeding programme as a whole. Which of the phenotypes are still in production and what you may have done differently to maximise your results. Looking forward to seeing what 2023 videos you have in store for us eager viewers
I have a bunch of cordyceps phenotypes that are currently being crossed on agar. I really refined the gourmet’s and my interest is going to be companion mushrooms- what ones fair well together in the same parameters 🙏🏻🍄❤️ stay tuned!
I wanted to go to school to become a mycologist yet they said I needed to get a doctrine which is alot of school facinating about peroxide I've done some crazy peroxide experiments as well. Thank you!
You can learn whatever you want! Check out Evergreen State College in Washington I believe they have an undergrad program for mycology - 🙂🍄❤️
Hey Gary, is it good when there are mated pairs? Will you be selecting from those regions? Or what are the benefits/disadvantages to selecting from an area that has mated pairs vs not?
Hi Gary! Thanks for posting all your work, I love watching and learning from what you’re doing. I would like to know if you use glass or plastic dishes? and if glass, do you sterilize and reuse dishes? If plastic, where do you source your sterile dishes from? Thank you 👍🍄
we use plastic dishes from wherever has them these days ha here is a link for some decently priced ones amzn.to/32QV4rX
Whats the ratio of h2o2 to agar here? Does it actually kill the unwanted contamination or just stall it? Do you think h2o2 could be used in a bulk substrate to help cut down on contamination?
Do you think you can do a video specifically on haploids?
do you label each bag in your fruiting chamber (M)1, 2, etc. and what is your method to keeping all of this data organized?
Whenever I rehydrate my grow boxes to initiate a second flush. I always add about 10% peroxide to the water
Peroxide kills the “Spores” not Mycelium, so Shouldn’t it been better to not add it Until After Germination??!
Hey Gary, I'm wondering how one can tell what regions of mycelium are "strong". It's very hard to see what you're seeing in the video, as all of the outskirt/periphery regions of mycelium look the same all around, rather than any one part looking different (or strong). Could you please describe what to look for visually? Is a stronger region denser, whiter, or have longer-reaching hyphae? Many thanks for the video!!!
From my personal experience and what I like to do in my mycology studies is to go straight to grain with my spores and grow the mushrooms out first then pick a fruit to clone and then isolate my mycelium that way.
ruclips.net/video/-GWXVGYzFs4/видео.html watch this one and maybe you will change your approach (it’s ok to do as a hobby but not ideal for scaling up)
@@FreshfromtheFarmFungi how do you know what phenotype you are taking out when you isolate before growing? This is the reason I go straight to grain, only enough for a few tubs. My spore prints are taken and stored in a clean room so very low risk of contamination. Seeing every variation before I start to pick my ideal fruit and then take samples to clone and then isolate.
@@allduhcheese6762 this makes sense for pheno hunting when you have a good, clean print - but when they are from the wild and easily contaminate it's not a viable option because the spawn will contaminate every time
@@FreshfromtheFarmFungi I hear that for sure. I do the same with my wild picked fruits as well. Take them into the lab ASAP and take a tissue sample, clean out any contam and then make bags or jars. I will then grow them until I see another good looking mushroom that meets my criteria.
How you work with the peroxid ? Do you mix it abd sterlise it ?
Well I would be doing this totally wrong. I would try to grab this big oyster ones that are clustered not the edges that looked cloudy. Then again it might not have looked cloudy in person.
You’ve moved since your video where you fruited in a plastic greenhouse, right? Would you still do a method just the same to check what the fruiting bodies of these mated pairs look like?
Yes I will be running these through fruiting and then cloning the strongest
Hey Gary.
What % peroxide do you use in your agar plates?
Mush Love
it was 3% - I normally don’t do this but thought it would be neat to test
Also a really good cross breed would be branched oysters with golden or pink cuz they get huge I don't know if you ever grown branched I had a wild species strain that were as big as dinner plates just awesome. And they seem to absorb or create I should say a lot of vitamin d which is critical to my health I wish I still had that strain but I had to stop growing mushrooms for like 2 years so I lost all my work. You wouldn't have a species of branched that I could buy off you guys would you?
I'm having a problem with some tubs. I'm only getting growth in the shrunken edges of the tub I've never had this before I've been growing mushrooms for 10 years I think I know what I'm doing I have 80% humidity 80° temperatures I mean I got enough light I found that white plays a very little part in most mushroom species but I mean I just can't figure it out. Are certain strains super dependent on co2 for pinning? I just don't get it I've never had issues like this in the past it's really really discouraging so much work I got like five or six tubs that are hardly growing anything.
How do you know what piece mycelium that you can salvage if there is bacteria
Also digging the hat
Is sabouraud agar worth using?
How can you tell if the mycellia have "eaten" or otherwise nullified a bacterial colony? I'm thinking that might be one to select for growing for reclamation or improving the environment.
You need to confirm with microscopy or other identification methods - I would recommend using sterile mycelium over reclaimed
I let my yellow Oyster spore syringe's rest standing up-right over night. I noticed today that a purple-ish colored sediment was collected at the base of the syringe's. Is this normal for Yellow Oyster spores?
Those should be spores my dude
Hey was wondering if it’s necessary to always use parafilm after each time u open one?
no but it helps keep them from drying out which is a big deal here in CO
I'm trying to start some wild reishi ( tsugae) from spores. I'm a noob and wondering why one would need to sector from one batch of spores. Aren't they all genetically the same? Being from the same mushroom. If anyone could explain or tell me of a video explaining this to a beginer I'd appreciate it. Thanks
Gary, I noticed that your agar plates have little to no moisture in them. Whats your process to manage that? Starting to get frustrated with the large accumulation of moisture in my plates. Keep up the great work you do. And I hope to take one of your classes in the near future.
watch this one ruclips.net/video/Rfd6F3Ac8OU/видео.html it’s all about temperature control - the condensation happens when the air inside the dish is warmer than the air outside it
I am also enjoying your informational videos, Gary. I think the lack of moisture has somewhat to do with the Denver weather as well. The humidity is very low there compared to mine in Florida. Finding that sweet spot (temperature)to pour can be tricky.😎
I'm novice, but I'll flip my upside down diagonally, and repeated flick it, not full strength!, and leave upside down, but I think that causes my fluffy mycelium to reach to the water...?? Lol like I said I'm new, but at least you can view what's growing.
How are you able to guess what was mated versus what was isolated? Thanks ❤
I would like to know as well! Hopefully I can find out. Have you found out yet?
What temperature do you normally incubate the dishes? define room temp if thats the case.:)
72F
Wisconsin accent?
Also luv your content. Mush luv ❤️
Buffalo/Colorado accent now ha it’s similar - thanks for watching and following along!
What should I consider when doing "strategic crossing of isolates for mated pairings"? What do you look for to make it strategic?
selecting for speed of growth, resistance to contam, pinning qualities, yield, co2 resistance, fruiting qualities, pins to flush time etc.
@@FreshfromtheFarmFungi rad, thanks! So when youre crossing isolates, how are you supposed to identify those character qualities ahead of time if theyre only haploid?
Breed haploid A out with haploid B, take note of the qualities in fruit, assume the quality exists in both, then breed haploid A again with Haploid C, and see if those qualities arent there anymore... Then you know those missing qualities were unique to Haploid B?
Lemme know if that makes sense
Doesn't peroxide also kill mushroom spores?
not sure - someone suggested this early on so thought I would test it
@@FreshfromtheFarmFungi I've heard about putting a 3% hydrogen peroxide solution into the agar after pressure canning and then it kills all fungal spores. Mycelium supposedly converts it to water pretty easy. Anyway, like the vids, nice and informative.
Edit: it's 3ml 3% peroxide to 500ml agar media after pressure cooking/canning. It kills all airborne mold spores but unfortunately kills mushroom spores as well. Doing mycelium transfers isn't a problem. Mycelium deals with it in a day or two and converts it to water and grows normally afterwards
Do your blades flake apart after flaming more than once? Always what to do about the black soot on blaes caused by flaming?
I use fresh blades between cultures or an induction heater like the one in this video it seems to help! ruclips.net/video/fmC38jxPbQc/видео.html
Will haploid mycelium fruit without pairing with other mycelia?
no it needs another haploid to be viable
You mean that the Petri dishes came with some contamination and only the peroxide is able to eliminate this? Aren't Petri dishes supposed to be sterile?
no if you watch the first video you can see - I inoculated the plates with spores, some got contaminated (4 of the blue oysters) I then put peroxide on two of them and left the other 2 to see if I could recover any mushroom mycelium Im doing a whole stitch together video on that separately (ruclips.net/video/nK8AghZs5ts/видео.html)
What do you mean hydrating the spores? I have not heard of this
when spores are left on prints they often dry out. Some spores have lipid bilayers some have thin layers and if they dry out they become unviable. It’s a good idea to hydrate them first though with water which allows them to germinate easier. This is why spores are not germinating all over at all times. they need water
Howd u get single spores onto that dish?
ruclips.net/video/GaO2q-mTvJg/видео.html watch this I explain everything
Need a light table
Watchout! That sucker might have teeth! What you feed that bacteria?!?
One more thing could you offer a brief definition of the various technical verbiage? Can be a tad tricky to comprehend without understanding the terms
any terms in particular? Ive been engrossed for so long I don’t know what is technical anymore ha
@@FreshfromtheFarmFungi I'm referring to the terms for the various spore types. I'm trying to learn how to make new strains with the cordyceps. But feel like I don't know how to ask the questions in my mind. Plz talk about ascospores vs the other types.
@@FreshfromtheFarmFungi endospores, ascospores, pheno type. Those kind of words that relate to mating the Cords. That's what I'm looking for. Thx man, I feel smarter every time I watch you work :)) #MushroomFantasies2022
I'm surprised about the way you work! I always pour, transfer and seal my plates very careful but bacteria/yeast is always a problem, even working in front of my flowhood and flame sterilizing with 86% industrial alcohol. I also use alcohol/bleach or ammonia in my hands. Where am I failing :(
I love your videos 8)
it could be many factors - hand positioning, breath, flow hood cleanliness. try leaving a plate open in front of the hood for 15 minutes it should come out clean. Next, try wearing some ppe - it might help. Also avoid crossing in front of the plate it might be blowing particles onto the surface of the agar from your clothing, skin etc.
Check your filter in the flow hood. Maybe it needs to be replaced? Also consider air currents and how they shift because of our movements. Always have the cleanest material in front. There should not be anything between the front of the flow hood and your cultures or new plates.
Use 70% alcohol. It takes longer to evaporate hence sterilizing better.
First!
nice!
when you run that blade through mycelium from one plate, then take another transfer from another plate without sterilizing the blade, you've just wasted all your time.
yes this is a critical mistake in this video - I usually do serial dilutions, germinate one spore per plate and change blades between each transfer.
Why not just throw the contamination away instead of putting peroxide to heal it, and just use clean non-contaminated substance,
this can work but often when contaminants are disturbed they release a plume of spores and it makes things worse. If you are very careful and precise it can work in a desperate scenario. I usually just toss them and try to repurpose the plates
That's a lot of contam bro
the cost of making videos and talking through. Overall I get about 1% contamination, the goal is definitely zero
I hate that RUclips keeps recommending your videos… This channel is totally mind numbing. The amount of talking about stuff that’s not necessary usually takes up half the videos…
This video could’ve been 3-5 minutes long with all the info you shared if you left out the rambling.
Aside from that, the monotone voice is unbearable to listen to. I’m not saying be all hyper like me beast or other “influencers” with those thumbnails that all look the same.
But, I can guarantee you that the amount of people you have leaving the video around the 20-60 second mark would change drastically if you shortened the video and talked like you are enjoying it instead of sounding like this is something you don’t want to be doing.
You would probably see a huge increase on people watching until the end.
Your stats aren’t looking great, but you obviously have the knowledge about this that not many others have. You just gotta change those couple things and your channel would grow insanely fast.
Right now the 2 most popular channels according to their stats are the channels with 3-15 min videos that have time stamps and explain everything in a fast and entertaining way.
Don’t take what I said the wrong way. I’m not hating or anything, but I’m just sort of pointing out some flaws that could be easily fixed and make your videos so much better!
Best of luck my friend
I appreciate the honest feedback. I mostly do this channel as a hobby and passion but as it gets more popular I should consider dialing in these things. Right now my audience retention is 68% for this video which is above average but can definitely improve. I will take this to mind when making future videos I just like to get my thoughts out there to the community however it comes off the dome hah 🙏🏻🍄❤️