Making Mushroom Grain Spawn in the Lab with Mushroom Liquid Culture
We're in the lab today. We're going to be doing some some work or creating some generation one grain spawn bags. So, the generation one grain spawn bag is basically a bag that will inoculate straight from like an agar wedge directly into sterile grain. Once that bag is colonized, we can create generation two bags. So, each of those bags is utilized to inoculate 10 more bags.
And those bags will be used for production. You can keep going to a generation three. So, one agar wedge can become two of these spawn bags. So, these bags were actually just inoculated about four or five days ago with a couple agar wedges. And we can see the mycelium starting to weave through the grain.
This is just a hard red winter wheat. You can use a variety of cereal grains. We like to use red winter wheat or white proso millet or milo. But yeah, this bag will get broken up into 10 new bags. So, one petri dish becomes 10 bags.
Those 10 bags can become 200. Each bag can be utilized to inoculate 20 to 50 production blocks. So, that 200 can become quite a bit of mushrooms. 200 spawn So, yeah. I'll put these away right now.
Kind of get set up. We have some fresh sterilized grain in the sterilizers. So, I'll just get myself set up real quick. Making sure I have uh some alcohol on these containers. These come in handy just for washing your hands throughout the process.
Easily between transfers or between handling bags. Just like to get a nice douse of isopropyl and and utilize it. We have some liquid culture that we just inoculated also with the agar wedges. So, you can kind of see the mycelium kind of growing through the liquid. This is just about four or five days old at this point.
And we can just use the magnetic stir bar in there to agitate the liquid or we could even just swirl it around like this. You can kind of see the mycelium is nice and developing really well throughout the nice media broth. We'll be able to extract this in just a few days. So, we just extract the liquid through the injection port. And you can use that to also inoculate your sterile grain.
Um We also were experimenting with these lids. And they have basically a place where you can connect your syringe and just uh suck the mycelium straight up out the jar without having to really tilt it. So, this actually came from a company called Agar Addicts. So, really cool jar lid. Um Yeah, seems to be working really good.
Written protocol
Southwest Mushrooms Genetic Library
The strains themselves, the same cultures used in this footage.
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So, these sterilizers come in handy for making like the generation one spawn bags. Then we can use the autoclave that's built in to make the generation two. Do about 80 bags a run and use use eight spawn bags to inoculate those 80 bags. And there we go. So, actually just give ourselves a good misting of the sterilizers.
Now, I'll just do a quick wipe down the inoculation area or our surface area in front of the flow hood. So, I'll go ahead and vent out the pressure. I'll just do it one at a time for now. And as soon as the pressure is released, we can just go ahead and open up the lid. And just do the same thing over here.
So, we're sterilizing about 50 lb of grain in each of these sterilizers. With three of them running, it gives us about 150 lb of fresh grain. And we can just use that for like I said, generation one bags. We also like to use these for making like liquid culture or agar media. All right.
Go ahead and close that up and make some room. I'll set myself up for a nice inoculation. Go ahead and these petri dishes have been examined for just optimum growth. So, we can just see that the mycelium is healthy. It's colonized across the petri dish.
We just have a multitude of varieties that we're going to be using. So, this is like the HE0. So, this is a nice lion's mane strain that has real long teeth. It kind of really resembles the or closely resembles like the bear's head or Hericium americana. So, it's really uh really good strain that I've loved growing for a while.
We also have a blue green oyster, which is a really really beautiful oyster mushroom. Colonized is oyster mushrooms in general typically take over your agar plates within a matter of days. So, this petri dish is only about like a week old. So, we can see some nice growth. Couple shiitake strains.
Nice shiitake right there. We have like another oyster variety. So, yeah. You'll notice just different different kind of myceliums depending on what the species or strain is. Like the pink oyster for instance will develop a pink hue to it.
So, like that's our pink oyster right here. You can notice a nice little pink hue to it. A real nice cottony growth. And really aggressive. So, strain that grows in a really high temperatures and is really forgiving.
Even if your growing environment might not be adequate or proper humidity, the strain still develops really well. The only thing is it just has a short shelf life. So, it's great for like drying and stuff like that and uh using it as a dried product. Or if you just know people that are ready to use them right away, then it's perfect. We just go ahead and unwrap the petri dishes off the parafilm and just set them back there.
Just go ahead and toss that aside. So, I'll go ahead and kind of label the generation one bags which with a sharpie that I have on hand. That way we don't get confused or any spawn mixed up. Have my scalpel nearby. Also something to sterilize my tools.
I really like this. It's just like a magnetic tray. So, everything sticks to it. It sticks to the table. Works works really well for keeping all the tools safe.
Go ahead and remove the lid. I'll just touch the scalpel right to the middle of the plate. And just I can just let it inflate with that sterile air. Just check it for a quality seal. And I'll just move that aside while I inoculate the next bag.
Usually 10 seconds or so does it. Now that the wedges are in the in the sterile grain, we can just break them off. And that'll actually break up the mycelium and allow it to spread it further throughout the grain. Now there's just an increased colonization rate. Make sure all of our moisture gets redistributed.
THERE WE GO. WE HAVE A NICE inoculated bag of grain. After the lab is done, uh we'll go ahead and place these in a in a section of the facility where they can incubate and go ahead and colonize. Then we can uh monitor them and bring them back in for uh further transfers. So just do the same thing here.
Yeah, I found that just breaking up the agar wedges uh that could be the benefits of using bags um for colonizing spawn just because you get the agar wedge breaks up into a tiny little piece uh a bunch of little pieces, hundreds of little fragments. And uh as it's shaken up and mixed with the grain, it's spread throughout the sterile grain increasing colonization times. And you can actually just do both of the bags at the same time. Usually what I do. Whoop.
I'll just have to replace those petri dishes, but we just had a couple fall. And at this point, they won't be used anymore. So they're basically uh contaminated. Contaminated. Um It's just not worth messing with at this point.
So we have more plates in the incubator. We can just go grab them and uh replace them. So that's why it's always good to make several uh plates for the same strain just so you have them on hand and back them up uh religiously. So.