A Day in My Plant Cloning Lab (rare plants & new experiments)

[00:00:00]Welcome back to the laboratory, my lab rats. Today, we have quite a few things to do. So, I'm glad that you're all here. The first thing we need to do is make some Monstera media so that we can subculture all of my Monstera Bulbasaurs. I believe this will be either their second or third subculture.

[00:00:19]I think second. While the media is in the autoclave, I'm going to subculture some roses. A few weeks ago, actually months ago, I went to Trader Joe's and bought a bouquet of roses, which I then took the nodes from and put them into tissue culture so I could clone the bouquet of roses. It was successful, so now I have to keep subculturing them.

[00:00:39]So, today will be the second rose subculture. And then the third thing I need to do, plant-wise, at least is subculture some African violets. I've been subculturing these inside of a Stella airbox exclusively now. I think this will be either their fourth or fifth subculture. I started this project last year, the end of last year. A lot of people start out doing tissue culture with a still airbox, which is awesome.

[00:01:06]But sometimes I do get negative comments that say, "Oh, you can't do TC in a still airbox. It doesn't work. Stop telling people to use a still airbox."

[00:01:13]So that's the main reason I've been subculturing these inside a still airbox, cuz it does work. It's just harder than using a flow hood. But obviously when someone's just starting out in a new hobby like TC, not everyone wants to go out and spend like $5,000 on a new gigantic flow hood. So I'm a big supporter of the still airbox.

[00:01:35]Originally, I thought that I would be putting some beonas into tissue culture today. Me greenery export had reached out to me maybe a month ago and asked if I wanted some free plants and I was like, "Hell yeah, I do." So, they sent me a Beonia maculada, the variegated version, which has been at the top of my plant wish list for probably almost two years. So, I was really psyched about that. They also sent me a Beonia Loon Mint, which unfortunately did not ship as well as the maculada.

[00:02:10]I'll partially take the blame for that because I left them in the box for like two extra days after they arrived. So yeah, I do plan to put both of these plants into TC. Probably the maculata first once it puts out a new leaf. The Lamoon Mint I will probably give some time to recover. I have them both in a rehab facility right now. Uh and by that I mean they're in a terrarium. Randomly yesterday, I came across a Reddit post by the person who hybridized the original beonia lmoon. And they posted this really incredible photo of a whole bunch of different beonas that all came from the exact same seed pod as the beonia lmoon. And you can see a lot of them actually look quite different from each other. I think sometimes because most of the plants that you can get at the store are clones, either propagated in tissue culture like this or just propagated asexually by taking cutings.

[00:03:06]We forget that plants in nature look a lot different from one another. For example, if you were walking through the jungle in Ecuador and you saw a monstera and then you walked 30 ft and saw a different monstera, they could look pretty different from each other, assuming that they're both seed grown.

[00:03:24]But when you get a monstera at Home Depot, they're all clones of each other.

[00:03:28]So like effectively thousands or maybe like even millions of people are growing the same exact plant. So we're used to plants like looking a certain way and not having a lot of variability. Okay, what am I doing? Let's get started.

[00:03:41]Today I'm going to be making 2 L. So I'm going to start with like 1,600ish milll of DI water, but you could also use distilled water as well. Ideally, you wouldn't really want to use tap water to make tissue culture media because you're going to have issues with minerals in the tap water. And really, the MS or your basil medium has all of the minerals that the plants are going to need. So, that's why you want to use either deionized or reverse osmosis or just distilled water.

[00:04:15]Since I'm making 2 L, I'm going to need 8.86 86 gram of MS. Oh my god, who needs the lab spatula?

[00:04:26]Not me.

[00:04:29]Now, I'm just adding 50 g of sugar.

[00:04:37]Next, we're going to need 10 ml of BAP, which sounds like a lot, but we are making 2 L. Monsteras definitely require a higher concentration of hormones than any other plant I've ever worked with in order to get good multiplication rates.

[00:04:56]And then I'm going to need to get the smaller pipet out because we only need 0.2 ml of NAA, which is our oxen.

[00:05:09]And it's just a tiny little amount.

[00:05:12]Now I'm just filling it up to the 2 L mark. Uh pro tip, turn off the magnetic stirer while you do that. The last thing I need to add is my AAR. And I'm going to measure that out and add it directly to my 2 L media bottle. I like to use for the Monster about 6 g of AAR per liter.

[00:05:34]Close enough.

[00:05:36]The last thing we need to do is just adjust the pH to around 5.7. So, I'm just adding a couple drops of pH up solution at a time until I get to approximately 5.7. And you always want to adjust the pH while the magnetic stirer is stirring. I'm at 5.74. So, I'm going to stop because that is pretty dang good. Oh my god, I'm spilling the media everywhere.

[00:06:09]So that is I forgot I spilled it everywhere. Okay, so that is our monstera media.

[00:06:29]Now I'm going to be subculturing my roses. But before I do that, I just wanted to show you cuz right next door I have my gymnocalism calacium. I'll just call them gymnos cuz I'm probably saying it wrong. But my gymnos seeds have germinated. A few weeks ago I got really into Palm Street while I was working on a video about tissue culture sales on that platform.

[00:06:54]Um, and I started just buying plants for fun. I don't know. I've had to delete the app like twice now and somehow now it's back on my phone again. But one of my favorite sellers that I found on there while I was working on this video is Epic Succulents and Cacti. He sells all of these imported gymnos from Thailand and they're these very bright, vibrant varieties that I think are really fun and I'm surprised that they're not more popular. I've bought a number of plants from him at this point.

[00:07:24]I got some gymnos. I got one called like jelly mutant that was kind of cool, but it had a lot of variegation. So, I was like, "Oh, I should graft this to another plant." So, I tried to graft it to my monstrous totem pole cactus, which is not like a plant you would typically use as grafting stock. This feels so wrong.

[00:07:46]So, I cut off the head of that cactus.

[00:07:50]Now, I'm going to cut off the butt from this cactus.

[00:07:54]I'm just going to line up. Oh, this looks so cool. Okay, I don't regret this. So, I'm just going to tape it to this cactus.

[00:08:05]Okay, apparently God looked down and saw what I was doing and decided that I had finally gone too far because my totem pull cactus now looks like this.

[00:08:17]Anyway, the reason that I'm telling you all this is because from Epic Succulents and Cacti, not only did I buy some gymnos, I also bought these seeds that I just showed you. This one is LB hybrid, pink diamond, baby pink, and pink panther. If you need a really easy first TC project to get your feet wet in the hobby, highly recommend germinating seeds on TC media. It's really easy.

[00:08:47]What I did was I added 5 ml of 3% hydrogen peroxide to a centuge tube, a 50 ml centuge tube. And then I filled the rest up with water and I added the seeds. Then I just basically left them in the solution for 24 hours. I rigged this up to just agitate the seeds in the solution a little bit overnight. I'm going to leave this running until tomorrow. And then when we come in tomorrow, we will plate these bad boys onto this media that I have over here. I left them on my orbital shaker, but you could also just flip the tube upside down and just leave them in the tube overnight. I've done that before, too. I plated them on seed germination media using a sterile pipet tip that I had autoclaved, but you can also just pressure cook it or put it in an instant pot. Um, and then using the pipet tip, I picked up some of that solution that contained the seeds and plated it on the TC media. I was in the flow hood obviously, but you can also use a still airbox for this, too. I have done it in the still airbox before and found it not that difficult to do. Okay, I fear I'm procrastinating. Let's subculture the roses.

[00:10:03]So, last time I subcultured these roses was May 31st. So, it's been pretty much exactly 4 weeks, which is the ideal kind of range where you would want to subculture your plants. More consistent subcultures is always going to result in more consistent and better growth for your in vitro plants. Oh yeah, also pro tip. I don't know if other people are doing this, but I'll just put like the whole kit of what I need into the autoclave.

[00:10:34]So like the tray, multiple pairs of forceps, and then today I'm going to be using scissors. So that's what's in here.

[00:10:43]If your trays are sliding around, I don't know if people know I do this, but I literally just like tape the tray down and then it won't move.

[00:10:53]So, what I'm doing is just removing any dead tissue that I see. For example, brown stuff like that, brown leaves.

[00:11:03]You really don't want any dead tissue in your tissue cultures. This expplant looks okay, but it's breaking apart like super easy. So yeah, I could really separate this into two different plants, but I'm going to leave them attached for now. And then the next time I subculture them in about a month, I will separate them.

[00:11:36]So there's our first rose plant. This is also by far the biggest one.

[00:11:43]A few of the other ones that I have growing like this are basically still nodes. And honestly, like what is going on with this one?

[00:11:52]So, like something like this, I'm literally just going to take and pop it into just some fresh TC media.

[00:12:01]Just like that.

[00:12:07]This one is just creating a bunch of callous.

[00:12:12]Idk. What to even say about that one.

[00:12:32]So, we have all of our roses replated onto fresh culture media. This one by far looks the best. So, all of you guys need to try a little bit harder. I'm just going to put these guys back under the lights so that they can continue to grow. I don't know why these are up here, but I'm going to move them.

[00:12:55]We have the roses back over here on the shelf. And now I want to take it right next door to look at the African violets. Now, you might look at these and say, "Laura, those look contaminated." When you don't subculture your plants enough, which is what has happened here, clearly the last time I subcultured was March 5th. It's now the end of June, so it's been officially just way too long. When you don't subculture your plants enough, you get buildup of ethylene gas, which is what is in these containers, causing this discoloration in our tissue culture media. Obviously, some of the containers look worse than others. And in fact, I do see a contaminated culture right here.

[00:13:41]That, my friends, is slime mold, which means this one's going to go in the trash. Honestly, the longer you spend between subcultures, the more likely you are to get contamination. So, I'm going to subculture these today to give them a new lease on life. But the positive is that the multiplication is looking really really good in a lot of these containers. I've got my still airbox set up over here. So I have the I don't want to point cuz I'm holding the mic, but I have the fresh media on the left and then I have a little jar of um isopropyl alcohol 70% that I'm going to be using for my forceps when they're not in my hands.

[00:14:23]So, I'm going to start with these six containers of Xplants. And then the reason I have more media is because some of these are taking up a lot of room in the culture containers. So, for some of these, it might actually make sense to split them into multiple different containers so that they can grow better and have better access to the nutrients that are in the fresh media.

[00:14:48]So, it's a little lower tech, but the same basic process applies for what we did in the flow hood. You just move the trash down here.

[00:14:59]Uh, which is just basically that we're going to take this out, put it into fresh media. That's it. So, I just loosened the cap of the container that contains the expplants. And then I loosened the cap on a container that contains fresh TC media. And then all I'm going to do is take my forceps, remove the expplant or a portion of it, and place it into the fresh TC media.

[00:15:29]Generally, one thing that can help when you're working inside of a still airbox like this is not using that much agar in your media. Uh, the less agar you use, the more money you save, lol. but also the easier it is to press the expplants into the media. As you can see, I'm having a little trouble pressing them in.

[00:15:50]There we go.

[00:15:53]So, that's it for our first container.

[00:15:55]You can also shove your trash into an old container of media. Now, it's contained.

[00:16:01]Okay. So, I'm going to loosen up the cup that contains my expplants. I'm going to loosen the container of fresh media.

[00:16:09]grab my forceps again and quickly place it into fresh media.

[00:16:15]Again, this would be way easier if our media was a little bit softer.

[00:16:20]And then I'm going to repeat that for the rest of that expplant selection that's in this container.

[00:16:37]You want to just do your best with it.

[00:16:38]It's never going to look perfect in the still airbox, but you can see we have good contact with the media, which is the main thing that we're looking for.

[00:16:46]All right, guys. So, here are all the African violets after our still airbox transfer. I think I'm drunk from breathing in all those alcohol fumes. Is that possible? Let me know in the comments. But yeah, they look way better than before. Time to go get the media out of the autoclave.

[00:17:19]Oh, you know how I'm not working with the scalpel today.

[00:17:50]For the most part, I'm just going to be opening up the containers and transferring the plants into fresh culture media, which is what I just made.

[00:17:59]If they have roots, then I'm going to cut the roots off, which made me just realize that I need to get a pair of scissors. This one does have roots, which I didn't realize at first. But look at that multiplication rate. Now, for other plants, this wouldn't be that impressive, but monsteras do not multiply at the rate that filidendrrons or alocacas do.

[00:18:23]Out of just this container by itself, it looks like we have five plants, which is a really great multiplication rate for a Monsera. Actually, I think I count six with that new shoot on the bottom. I did such a bad job transferring that one, but whatever.

[00:18:41]So, there is our first one.

[00:18:44]We'll do this one next.

[00:18:49]The variegation on this one is really pretty.

[00:18:54]My hands are super shaky today from drinking coffee. I normally don't drink coffee in the afternoon.

[00:19:03]Goodbye to the roots.

[00:19:10]I was talking to some people in my Discord yesterday about future video ideas and someone mentioned that in Japan they genetically modified a Phelinopsis orchid to have blue flowers.

[00:19:25]So the plants got approved for commercialization in the United States last year. When you're dealing with any sort of genetically modified plants, there are a lot of regulations behind whether the plants can be commercially sold or not for a lot of good reasons.

[00:19:43]Uh, but the orchid did get approved. So, I think they're going to commercialize it in the United States. At least, I really hope that they do because I really want one. I did find a guy in Canada selling them. I don't know where he's getting them from. I guess from Japan. So, I messaged him to see if I could purchase one, but they haven't gotten back to me yet. But I would really love to have one for the video itself. So, if you're watching this video and you have a hookup that can get me a Japanese blue jean orchid, I would really like to get one.

[00:20:34]This one I'm literally just going to pick up and stick into a new container of media. I try to fill the media containers a little bit higher when I'm working with monsteras cuz obviously they're just larger plants. for like African violets and stuff. Usually the rule is like one finger width is how high you want your media in the media containers. Yeah, I could have even filled these media containers up a little bit more. So, I'm pushing them all the way to the bottom, but I wish I could even push them a little bit deeper. This one looks kind of messed up.

[00:21:07]This would be an example of a plant that if I was a commercial laboratory selling plants for money, that would be one that I just throw in the trash. But we'll keep it for now and see if it does anything weird. This one has very pretty variegation. Although it looks like the new shoot, at least one of them, doesn't have any variegation at all. It is the risk you take when tissue culturing plants that have chimeic variegation.

[00:21:38]Just cut that root off.

[00:21:40]The next time I subculture these in another month will be when I separate all of these shoots.

[00:21:47]There's that guy.

[00:21:50]Oh, we have a contaminated one. So, here's an example of one that I'm not even going to open. You can see this kind of sliminess that travels across the media here. That is contamination.

[00:22:01]So, ideally, I really don't want to open this one up inside the flow hood. And I'm actually just going to dispose of it.

[00:22:10]And there's my mountain of plants that I've been building. All right, guys.

[00:22:13]That's going to be it for today. I want to thank you all very much for watching.

[00:22:18]If you can help me find a blue jean orchid, as I mentioned before, hit me up. My email's on my website. Also, if you'd like to learn how to do tissue culture at home, I will link a couple of beginner tutorials that I have on my page. I know I'm obviously in a laboratory today, but it is possible to do at home. If you need supplies for doing tissue culture, check out my website. We've expanded our product range a little bit lately, so if you haven't visited plantsandjars.shop in a while, you might see some new things in there. Mhm. Okay. Thank you guys for watching and I'll see you in the next video. Bye.