Shaping Fire Ep. 64 - Feminized Seeds and Female-Only Breeding with guest Ryan Lee

As more and more people begin to grow their own cannabis, the popularity of feminized seeds has risen remarkably. As new growers come to understand the inconveniences associated with using regular male female seeds, feminized seeds have become an increasingly attractive option. If you are a listener unfamiliar with feminized seeds, these are the three most common reasons people choose feminized seeds. Number one, in most cases, a patient is legally limited to how many plants they can grow, so each plant is exceptionally valuable. Generally speaking, cannabis seeds will germinate and become plants that are 50% male and 50% female. So when you pop your regular seeds, you should expect about half to be female. However, that 5050 only plays out statistically over thousands of seeds, so it is entirely possible for you to pop six seeds and have four or five of them be male. It is exceptionally disheartening for a patient to discover that so many of their plants are male. When they need female plants to make their medicine and get relief. After all of the soil and nutrients and care and love the patient has given the plant, it is a significant disappointment to have to destroy it because of this. Many patients love feminized seeds because it removes the risk of cultivating plants that end in wasted effort. Number two, Cultivators who are growing more plants than a patient but less than a commercial cultivator often need to make clones to extend what they can do with a small number of seeds. This often means going through the arduous process of popping the seed, letting it grow, taking clones, flowering a clone to determine the sex of the plant, and then planting the clones that prove female while culling the males. That process takes some time. It is far more convenient to pop a female seed and just start cloning, so confident that the new clones are all female. Number three, commercial scale growers are now embracing feminized seeds too. Commercial cultivators have a choice of either to use clones or feminized seeds. Fem seeds are easier to store and have that vigor that comes with a seed instead of a clone. For now, though, feminized seeds are a bit more variable than clones. So many still use clones. No doubt there are plenty of great reasons to grow male female regular seeds. This episode is focusing on the benefits and challenges of feminized seeds and female only breeding though, so we won't be talking about the reasons we like regs. This is an episode Exalting the Female if you want to learn about cannabis health, business and technique efficiently and with good cheer, I encourage you to subscribe to our newsletter. We'll send you new podcast episodes as they come out, delivered right to your inbox, along with commentary on a couple of the most important news items from the week and videos too. Don't rely on social media to let you know when a new episode is published. Sign up for the updates to make sure you don't miss an episode. Also, we're giving away very cool prizes to folks who are signed up to receive the newsletter. There's nothing else you need to do to win except receive the newsletter. This month, regenerative farming nutrient company Everflux is giving away their full line of products to one lucky subscriber. To the newsletter you'll receive a full sized bottle of their bioflux fermented plant Booster, their bamboo Wood vinegar biostimulant, and a big bucke of Terraflux, their infused biochar blend.

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Make sure to listen to their commercial during the first break today and go to shapingfire.com to sign up for the newsletter and be entered into this month's and all future newsletter prize drawings. You are listening to Shaping Fire and I'm your host, Shango Loess. Today my guest is Ryan Lee. Ryan is a professional cannabis researcher and plant breeder specializing in cultivar design and the inheritance of therapeutic compounds in cannabis and including cannabinoids and terpenes. His background training is in neuroscience and studying the endocannabinoid system with a strong focus on genetics. His post grad work at the University of British Columbia is all about plant breeding and biotechnology. Ryan Lee is founder of Chimaera Genetic Resource Management and the Chemovar Corporation. Chimera is essentially a seed bank and a cannabis germplasm firm operating in Europe.

For over 20 years.

ChemoVar Corp. Is based in Canada and focuses on breeding specialty cannabis varieties, laboratory analytics, cultivation, consulting, cultivar selection and germplasm sourcing, licensing and importation for Canadian LPs. Chemovar has guided clients worldwide to navigate international regulations surrounding the movement of cannabis genetic material. In addition to sourcing importing cannabis starting materials as the foundation for corporate cannabis production, academic research and and large scale breeding programs focused on next generation cannabis cultivars. Today we're going to talk about the creation and genetic makeup of feminized seeds and we're going to do a dive into female only breeding too, which is super interesting. Stick around.

Welcome to the show Ryan.

Thanks Shango. Really appreciate you having me on.

Excellent. Glad to have you here. So let's start with some vocabulary, right? Because we're going to go through a lot of the various myths around feminized seeds today, but one of the first things is like pretty much a comprehensive confusion between the vocabulary of feminized seeds and self seeds. So will you clarify and tease those apart for us?

Yeah. Okay. So feminized seeds is a word that really, I think was kind of coined or made up in the cannabis industry. It's not a word that we use in botany, but what it really means is seeds that you plant and they grow out to be all females. And, you know, this discussion is really going to come into some words that people might have heard when talking about buying seeds. Different cannabis seed types. So we have monoecious seeds or dioecious seeds, and hemp tends to be monoecious, which means that we have both male and female flowers on the same plant and drug. Cannabis is kind of at the other end of the spectrum where we have male and female plants occurring on separate individual plants. And that comes from the root word, the Greek word oikos, which means home. And so monoetius means both of the sexes reside in one home, whereas dioecious means the sexes reside in two separate homes. The word feminized really is kind of a more user friendly term for what botanists called gynoecious, which means the female house or all the seeds in the population being from the female gender.

Wow, that's actually really cool. So if feminized seeds are gynoecious, where in that model does a self seed live?

Yeah, so a self seed is a specific type of feminized seed where the pollen donor and the plant that actually holds the seed, which we call the seed mother, are genetically the same plant. And so it's the most severe form of inbreeding one can do on any given species. And it's something that's quite unnatural for cannabis. Cannabis in its natural state of dioecy, where we have, you know, if you were to come across a wild stand of cannabis somewhere, walking through the valley, you know, a valley, Tibet or somewhere, approximately half of the plants would be female and half of the plants would be me, would be males. So it's a natural condition for any plant to leave its offspring that it has to mate with another individual of the same species, kind of like humans. And that's really quite rare in the plant world. And it has a whole bunch of consequences for genetic diversity and a whole bunch of other things that we're going to talk about over the next hour. But for example, tomatoes, they naturally self pollinate, so it's a natural state for tomato to, you know, for a bumblebee to come along and play with the flower on the tomato plant. And that causes a self fertilization where the male parts of that individual flower interact with the female parts, or the bumblebee transfers the pollen from one part to the other, and that creates a fruit in which there are seeds growing. And so if you, you know, if you buy heirloom tomato seeds off the Internet and the company sends you a package of seeds, and you take those seeds and grow them into a full plant, and you harvest the tomatoes and you take the seeds and you plant them again the next year, the plants from the next year and the fruit from those plants from the next year will be near identical, if not identical, to the seeds that you grew out on the first year. And if you were to take those seeds and continually save them and plant them in your garden over the next 10 to 15 years, you could expect that those tomatoes are going to be the same every time. Whereas, you know, we've come to see that's not the case in cannabis. And that really has to do with this natural state of breeding where every individual must interact with another individual to create progeny, kind of like humans.

So, so are self seeds all female?

Yes. Well, not necessarily, because you can actually self. We have a little trick for making female. So maybe I should talk about first how a feminized seed is made. There's a couple of different chemicals, but the one that's most commonly used is a chemical called silver thiosulfate. And silver thiosulfate blocks one of the five major plant hormones. And when that happens on cannabis, when you take a female cannabis plant and you induce it to flower and immediately spray it with silver thiosulfate, the silver ion blocks the ethylene in that biological pathway that leads to female flowers. And the consequence is that this female, genetically female plant will produce male flowers and thus they will produce pollen. So that's a trait of a male plant to be able to produce pollen. And it's very atypical for a female plant to be able to produce pollen. The difference being between male pollen and female derived pollen is is that the female pollen has no Y chromosomes. It has no uniquely genetically male contributions that are being passed on to the next offspring. And so what it allows us to do is mate one female with another. Or in the case of self seeds, we can mate one female plant with itself. And so self seeds are really, we do see a little bit of them appearing on the commercial market, but it's really a breeder's tool for doing what we would call severe inbreeding.

Right on. So the only time that I have come across, you know, in my own hands, S1s self seeds are I'm a big fan of Caleb over at CSI Humboldt, and he was on the show and he said that one of the reasons that he likes self seeds is because it kind of gives you a decent snapshot of.

Of.

Many of the different fenos that are in the parent plants. So whereas with regular seeds, when you get them, you'll have a differentiation of fenos, but you may not get everything. That's to be kind of sloppy with my vocabulary in the line of that parent. Whereas when you have an S1, it kind of creates. Normally we go for seeds that are all the same. Right. So that. So that all the seeds you get out of a pack are kind of similar and similar to the breeders cut. Right. Which for many people would consider like the gold standard. So you've got your breeders cut and then you try to make seeds that exemplify the breeders cut. But S1 self seeds, people seem to kind of go at it in a different direction. Whereas I've got the Skittles S1 that Caleb made, and I know that if I had enough packs of that S1, I only have one, but let's say I had four or five, I would have kind of a snapshot of all the variety that's in the skittles line. Do I have a general accurate understanding of that snapshot of the genetic line? And if not, where am I off?

Yeah, well, I'm going to say yes and no. Okay, fair enough. And so an S1 is what we call a recombinant generation. And what that means is I have to, again, I have to back up a little bit and to try to explain this. And we'll use humans as a way to do that because I think it's easier for people to think about sex in the terms of humans. Everybody has a mother and a father, and a lot of people have a brother or sister. So if we imagine that the mother is the ideal type, or as you called the breeder's cut, our idea is that we want to make. We want to mate her and have the progeny end up being like her. Right. And so in plant breeding, and really in all breeding, we have this basic tenant which is like, begets like. And all that that means is if you take a plant that has blue petals, it's more likely that the offspring of that plant will also have blue petals. Okay. But as we know from, you know, our experience with humans, you don't look like your mother or father. You might have some of their traits, but you're not an identical replication of the mother. Right. And so typically when people have made cannabis seeds, what they do is they look through a population of any type. Maybe it's a land race, maybe it's a seed pack they bought from Holland or California, and they grow these seeds out. You select an ideal plant that is, you know, excellent or good on a whole series of combinations. And if a breeder wants to then make seeds from that plant, you have to go looking for a pollen source, or traditionally you've had to go looking for a pollen source. And so what people do is they take their own favorite line and they use that plant to pollinate their new selected mother plant. Well, the problem is, is that, you know, following the like begets like tenet that I mentioned earlier, we're now introducing trai from a father type that might not be identical to the mother type. And there's one, you know, the easiest plant, the plant that would be most like the mother plant would be the mother plant itself, if we could find a way to produce it or to make that plant produce pollen. And so we use this chemical trick where we spray this ethylene blocker on the female plant when it's entering flowering. And that allows the plant to produce male flowers from which we can harvest the pollen and then self pollinate the plant. But still, we have to remember that that mother plant came herself as a hybrid from two different plants, maybe those plants were almost identical. And if they were almost identical, if we self the plant, that those self seeds would be very similar. Right? They might, they might even all be identical to the mother plant. But if the, if the mother plant that we're using is herself a hybrid, meaning she has, you know, genetics from two different types. When we, when we self that plant, it's going to expose all of the genetic combinations that exist within her genome.

And that's exciting, right? If you, if you're a pheno hunter, you know, if you're looking for unique traits. That sounds exciting. Now if you're looking for consistency in seeds, well, then you'd want the first version. But the idea of being able to see all of the wonderful varieties also sounds fun. Depending on what your goal is.

It really is. And typically, traditionally in the plant breeding world that, you know, that pheno hunt is that you're talking about, it really has been the goal of the breeder, the breeders goal is to put out, you know, as a commercial seed buyer, you want all the plants to look the same, or I would think that you want all plants to look the same. You know, you might be interested in looking at the variation, but I think for most people that are buying commercial seeds, they expect, like you said, a representation of a described plant, AKA the breeder's plant.

Yeah. Most people want consistency or what a lot of people call a stable line. Right. Whereas people will say, oh, no, this is a breeder breeders pack, because there's a lot of variety for you to fino hunt in.

Yeah. And that's kind of. I think that honestly, breeders have been able to hide behind that fat, that type of marketing for, you know, the last 30 years. And breeders have typically sold the variation as a positive. When I think that really it's a.

Feature, not a flaw.

Yeah, exactly. And that's, I think, you know, that's partly part of the reason, is that a lot of the people working with cannabis are, unlike Caleb, are not real plant breeders. They're just people making seeds to sell for money. And plant breeding is a skill and there's an education to it. And you have to learn the laws of how plant breeding and biology and meiosis work in order to be able to use them as tools to your advantage in creating a true breeding line. Self seeds are a way to get to that true breeding line, but they in and of themselves may not be a good way to produce stable seeds. That doesn't mean that they don't have their use, but to me, selfing is a tool in a breeder's toolbox. It's not the end result. It's not the end result.

Fair enough, Fair enough. So let's talk about the mechanics of using silvertheosulfate. One of the things I like about our conversations before today is that, you know, you've got the education and you are a scientist and, you know, in my experience, you describe things pretty well. So would you take us through the process of using silvertheosulfate to actually make a femini seed? I know this may seem awfully basic for you, but most of us can't get a good description anywhere because people describe things kind of sloppily. You know, even Caleb, right, who you and I both agree is a great breeder, when he was on the show, he's all like. He's like, man, I kind of just do it, you know, and. Fair enough, Right. So his, his expertise is making seeds, not necessarily, you know, Teaching it. But. But I'm gonna put that on you. So.

So.

So, like. Like, once we've scored some silver thio sulfate, you know, also what, you know, people call it feminization spray sometimes. Like, what do you actually do with it?

Okay. Well, I think it's important to understand how it works. So you don't need to use the silver thiosulfate. The thiosulfate component is kind of like a buffer or a transporter to help bring the silver into the plant. When the first researchers were looking at these things, I guess it's important to say, too, there's five major plant hormones. In humans, obviously, we have many, many more. But in plants, there's about five hormones that regulate most of the dance of life of the plant. And one of those hormones is ethylene. I think most people know the consequences of ethylene by taking a green banana and putting it in a paper bag and kind of cinching it closed, and the banana will go yellow. The reason is the paper bag is trapping that ethylene gas, which, in the case of bananas and many types of fruit, it causes ripening in flowers. It causes flowers to stay open. In the cut flower industry, you'll actually see flower producers typically all over the world, but they're shipping flowers from, say, Costa Rica to California for the market. One of the tricks to slow down the ripening process is to spray silver thiosulfate on the freshly harvested flower buds. That blocks the action of ethylene, stops the ripening, or slows the ripening process, allowing for a longer harvest window, you might say, or a longer transportation window. It just provides shelf life stability. So plant people spray these types of chemicals on plants. And I guess a researcher was trying to look at the effect of silver nitrate, which is very much like silver thiosulfate. They sprayed it on the cannabis plants, and they realized that this action would turn female plants phenotypically male. The female plants would produce male flowers. And it does that by blocking the action of ethylene at this ethylene receptor, much like Narcan does with the opioid rescue drug. It blocks the receptor and stops the action of the fentanyl of the heroin and essentially revives the person. So silver thiosulfrates does much of the same thing, except the end result is that the plant, about two to three weeks later, will start producing male flowers rather than female flowers. And we then harvest that pollen, or we put that plant in the same room or a pollination chamber with either another clone of genetically, the Same plant for making self seeds. Or we could put another plant of a different genotype in the same chamber as well to make hybrid feminized seeds.

So you'd put another female in with your female that you are applying the silver thiosulfate to?

Yeah, we call that plant the donor plant. And so the donor plant essentially becomes your, you know, air quote males, your air quote male plant or your pollen donor plant. And any other plants that you want to receive that pollen, you essentially put them in the same chamber, the same grow chamber, and they will also be pollinated from that one pollen donor plant. And so you can, you can make self seeds and you can make hybrid seeds at the same time.

For home growers who may not, who want to give this a whirl, but they may not have, you know, dedicated seed chambers and enough room for multiple plants. Would it be legit to spray the silver thiosulfate on say for example, lower branches and then just let the, you know, have a fan in there so that the pollen goes around the tent and then self pollinates the female flowers that are up higher in the plant?

Yeah, you can do it that way. I think most people that are playing around with breeding at home, they don't really need. I mean that could potentially produce many hundreds or even thousands of seeds. I think for most home growers, you know, 20 or 30 or 40 seeds is probably sufficient to play with. So yeah, you could do it. You could spray one branch if you wanted to, but then you risk pollinating the whole plant. It's probably wiser to spray a small clone that you can isolate from the rest of the plants. And then you, then you can collect the pollen from that newly male plant or the pollen donor plant and then you can just take that pollen and either apply it by hand or with a paintbrush to the lower branches of your choice.

Is the only downside to pollinating the entire plant just that it's gonna decrease the potency of the smokeable flower? Because as far as I'm concerned, if I was gonna go to all this frickin trouble, I would want to pollinate that whole donor plant so that it makes the most seeds possible for the labor that I'm putting in.

Yeah, you can, but it's again, it's not a very, you might want it, you know, in a small plant with say four or five large main colas, you could really produce a few hundred seeds of four different types. Right. If you put a different pollen donor on every branch. So again, I think for most home growers it would be more efficient to just pollinate single branches rather than the full plant. Because even if you just wanted to, if you say you're growing a couple of plants outside, you could just pollinate one branch by creating a small pollen. We call them pollen bags, and they're essentially just little parchment paper satchels. But you can put pollen in that and then tape over. You know, tape the end, the open end over a bag and it's white, so light still gets through it. Because it's parchment paper, it still breathes. And that way you can actually create seeds on one plant, or, sorry, on one branch of one pollen type without pollinating the rest of your plant.

Let's, let's talk about that. Let's talk about that pollen bag for a second. I'm familiar with taking the pollen and painting it on with a small paintbrush. Right. And that's fun and easy to do, these pollen bags. So if the pollen is in the bag, the pollen can't go through the parchment. That's good. And you're going to put it over the branch and you're probably going to tape it off to close the seal on the far end. What do you do, like, shake the bag once a day? Because I'm just imagining the pollen sitting in the, in the lower part of the bag.

Well, the lower. Yeah, it's. When you make the bag, it's in the lower part of the bag, but you turn the bag upside down as you put it on the plant. Right. So it kind of comes. You know, I would think of it like a sock being put over a branch.

Sure.

Right. The, the pollen is down at the toes when you're starting, when you're, when you're creating the bag, you're making it. But when you put the sock on the, the branch, so to speak, the bottom becomes the top, and you just cinch it with a zap strap or an elastic bag or a tie or whatever you want to do.

But you'd still want to, you'd still want to, like, jostle the bag occasionally to kind of re. Remove the pollen around. Right. So you get more chances to pollinate after the very first attempt. Right.

You can, it's, it's probably not necessary. I mean, when you're, you'd be surprised how many pollens or how many pollen grains fit into a, you know, for example, a teaspoon of pollen. So if you were to create one of these bags and put a teaspoon of pollen in it, there's a more than enough pollen to Saturate the entire branch. There's a great picture of these, actually on Rob Clark's book Marijuana Botany. Anybody can find that picture online. From the front cover in one of the four corners, there's a picture of a cannabis plant with four or five different branches on it with the pollen bags.

Okay, so if we follow this strategy of using sts, this silver thiosulfate.

What.

Are our percentages of success? Like, what's the likelihood of there being a male seed produced in this? I mean, it's less than 100%, right?

Yeah. I think the true answer to that, Shangha, is that we don't really know. I would argue that 99.99% of the seeds that are produced from these combinations are going to be genetic and phenotypically female. Now, we all know that there's anomalies in gender expression. That's a big topic about today, whether men are men and women are men or whatever. But there's definitely a small subset of the population where the genetics just don't match the phenotype for whatever reason. And those things can. Yeah, they probably can happen. But I think that it's much more likely when you find male seed in the, in a female, in an all female seed lot or a gynosis or feminized seed seed lot. It's more likely that that male is derived from a pollen contamination event back when the seed was making seeds or the plant was making seeds, rather than a female female breeding resulting in a male seed.

That, right, that right there is some good insight. I mean, you're in the scene, you know how often. I mean, it's not like it's incredibly often, but it happens often enough that somebody plants a feminized seed and they get a male and they totally lose their shit. And it ends up being blamed on the feminized seeds at a hole, when really it's probably an accidental pollen particle that contaminated in the breeding room.

It's much more likely. And like I said, there are anomalies everywhere in nature. Right. I was in Morocco with Rob Clark a few years ago and we were walking through these fields with thousands of plants in them, and we found a plant that had male, female, it had male flowers. But for every other discernible characteristic, this plant could have been a female. It was what we called a very female male. It was a heavily female trait male plant. There are some subtle differences between males and females. You know, for example, often, and this is not. It doesn't always hold true, but often when plants go into. If you Grow regular seeds out, dioecious seeds, and you put them into flowering, often the males, will they become taller right before you go into flowering. And you can think of that like an evolutionary trait, kind of like corn. You know, corn, the tassel that drops the pollen is at the top of the plant, right. So when that pollen falls, it's falling towards the female reproductive organs below. And so what cannabis does is it shoots up maybe an extra foot and so that the male flowers are physically above their female counterparts. And when they open the male flowers and the pollen falls down and yeah, it's caught by the wind, but it's still blowing slightly downward. Right. So it's an evolutionary advantage for the males to be slightly taller. And also that stretchiness that allows the male to stretch out and become taller. Also the same aspect happens to the flowers. And so a tight floral cluster really becomes much more loose and open with male plants than the female counterparts. And that allows the male. They kind of look like little grapes when they come out of a male. When they come out of their pre flower state, they drop and open and that, that loose, it's called like a panicle. And that loose floral structure, it kind of gives freedom for the fear for the male plants to swing around and be captured by the, the wind which jostles them and releases the pollen. When you spray STS on a female plant, you'll notice that the plants don't look like even though they're male flowers rather than female flowers on the plant, the rest of the stature is quite stocky and short and unlike a natural male. So you might have to, you might have to break up the female or the male flowers by hand, for example, to release the pollen.

Wow.

All right. So I've got a much better understanding of how to use silverthiosulfate to actually make the femini seeds. And what my strategy would look like before we start talking about accidental stress causing a female plant to produce male flowers that have pollen in them. You know, people have told me that, oh, there's lots of different ways to make feminized seeds. But what, you know, in my research, I pretty much find there be there is silver thiosulfate and then a bunch of rumor. So is there another scientifically reliable method to create feminized seeds other than using sts?

Yes, there are other hormone blockers. STS works the best. I think what you're really referencing is there's a couple of techniques. One of them was called rotalization, which is really not a good technique. And Dutch Passion, who first commercialized the feminized seeds also used a pretty suspect technique when creating the first feminized seeds. That really led to feminized seeds having a bad reputation. If you remember before I said like begets like. And all that means is that if you take a plant and it has any given set of traits, those, those traits are going to be likely to be represented in the offspring or more likely to be represented than the offspring. And so the way that the initial companies made these feminized seeds is they took plants that would say, Normally flower in eight to weeks, and they flowered them for 12 to 14 weeks. And when plants get way past sexual maturity, some of them will start producing male flowers. And so that those male flowers, obviously you can harvest that pollen and then also use that pollen to create new female seeds. But the problem is, is that those plants were, you know, some people would call them unstable. We, I call them intersex. It's so remember at the beginning of the episod, we talked about dioeciousness versus moneyciousness, where you either have plants that have separate sexes on separate plants, or we have the other end of the spectrum as both sexes on one plant. And so I would think of that like a, like a spectrum or a continuum where that one end we have complete intersexuality or monoecious plants like hemp that are, you know, they produce many, many male and many, many female flowers. And that doesn't matter because we, you know, with hemp, we want it to be seeded. Usually they're harvesting the seeds to be sold. But that trait in a drug, cannabis plant is really not a good characteristic because you can have a plant spontaneously produce male flowers and pollinate the entire crop or at least that region of your greenhouse or grow room. And that really obviously devalues the, the selling price of the cannabis if it's full of seed. So what these Dutch companies are predominantly Dutch companies did, is they used plants that would naturally produce. Keep in mind, this is before people realized that you could use silver thiosulfate. And so these plant breeders would harvest the pollen from these intersex plants and use that to fertilize their females or to pollinate their females. And those plants would then inherit that intersex or hermaphrodite characteristic. And so the first feminized seeds that were available on the market, we had a lot of reports of Hermes or more appropriately, intersex plants in the seed lots. And I think that that's one of the real reasons that feminized seeds kind of have A bad reputation on the market is because those early lessons, they left a deep impression in the culture. And the truth is, when you're taking a plant and you're selfing it, you're reinforcing any characteristic that is within the plant. So if you're. If your chosen mother plant has a slight tendency to be intersex or to go intersex, you know, on a. On a given set of stresses, ph, light cycle, you know, whatever it is, those stresses are likely to create a trigger. Right. To induce those male flowers to appear. And you really don't want to use. When you're selfing a plant, you don't want to reinforce that type of trait.

Wow. Yeah, totally. And so this is so interesting because this actually answers my next question I was going to go to. Because as we know, some cultivars, when you put them under stress, they will throw male flowers. And you know that for most of us, that's a negative happenstance. We don't want our female plants to throw male flowers. But then some people are like, oh, sweet. You know, I was hoping to get these flowers to smoke, but now they're feminized seeds. So now they've got this different value. And I was going to ask you, are those feminized seeds, you know, equally as valuable as the kind that we make using sts? And I think the answer would be.

No is because, no, they're much less.

Valuable, much less stable, and they're gonna. They're gonna include that likelihood that they will hermy or go intersex on you if it gets any kind of stress. Maybe not even a lot of stress.

Yeah. And, you know, again, I think the argument could be made that for a plant breeder, maybe using an STS technique or that type of pollination technique is actually a good way to remove the intersex plants. Because if you, you know, if you took a plant that had slight intersex tendencies and you selfed it and you grew it the next generation, any of the plants in the next generation that don't have that tendency, you know, it's possible that those plants have removed that given trait from the mother plant. So, you know, again, in plant breeding, there's no hard and set rules, but you definitely don't want to be releasing seeds like that onto the marketplace where a grower is then going to grow them, expecting that they're all going to be 100% female.

Yeah. And that's one of the challenges, too.

Right.

With the seed market as it is right now, it's so easy for People to make money selling seeds that, you know, you get folks that they take their, you know, favorite strain and they hit it to a bunch of hype strains and then they put the seeds out. Whereas, you know, more of the breeders that I respect, they're actually going to work that seed group to, you know, stress them and pull those out. So those traits are less likely to exist. And I think that's probably the real big difference between, you know, pollen chuckers and people who I give like, you know, the capital B breeder too is whether or not they put that extra effort in to really stabilize and pull out a lot of those unwanted traits.

Yeah, well, and that's, you know, that's just something that again, because of the, you know, plant breeding is typically done by nerds in greenhouses. It's not really a celebrity seeking job, but for some reason in cannabis it is. Right. And that's kind of a weird little commentary on the cannabis industry. But you know, in the real plant breeding world, as much as 75% or maybe even 90% of the crosses that you make don't, aren't, they're not successes. Right. Or they're not the ideal with plant breeding really in my mind should be. Well, I guess there's different goals in plant breeding. Like you said, one goal might be to replicate the breeders idiot type or the ideal expression or the breeders opinion of what that ideal expression is. And you want to capture that in the seed. Right. But there's different, you know, you might have an ideal plant like, I don't know, let's just choose one. The Girl Girl Scout cookies for example. I mean it's a great plant. It's super. It's a variety, it's, it's really high THC content, it's got a lovely flavor profile but it's not very big. So you might want to breed that plant to produce about twice as much as it does. And that takes time and effort and a lot of crosses and experimentation. And realistically, 90% of those breeding experiments are going to be dead ends. They're not going to lead to that next generation or what we would call an elite plant where you're raising the quality to the, to the, to the producer, not only the consumer, right. It, it produces more, it's more mold resistant, whatever, whatever the fix is for that particular line.

So before we go to our first commercial break, I want to go on a quick little fishing expedition with you. So at last, Emerald Cup. I was, I was hanging out in the Dragonfly Earth medicine tent and they were talking about how on their farm up in Canada they make. They have some plants that prod female only seeds without STs. Because they were saying at the time that they're not fans of using that chemical for whatever reason. And I'm all like, so how do you have female seeds naturally coming off a plant? And they said that their best guess is that they heavily green mulch with nettle, which is high in hormones, and that when this plant breaks down and it gets watered in that, at the very least the lower branches self pollinate, I guess is the idea. And then the plant produces only female seeds. So I'm curious to know, does this hit any triggers? And I want to apologize in advance to Dragonfly if I screwed up their story because maybe they don't produce male flowers. Maybe the seeds just come out as female. But I guess the point I'm trying to ask about is, is there a way that adding hormones to the root structure should change the gender of the seeds?

I love Josh and Kelly. They're great, really super nice people. But as a scientist, I kind of. That, that does trigger me a little bit. Yeah, I don't think that that's, I don't think that's likely. I mean, it's, it's. We, we have, listen, in the, in the cannabis industry, we have this long history of speculating and coming up with really lavish ideas. I mean, you know, people were driving stakes through the stems of plants and stuff 30 years ago to try and increase the resin content. We've. Humans have done all sorts of crazy things, but I think that scientifically there'd be. There's a pretty big gap between saying if we put these hormones on the root system, they get absorbed by the plant and they only have an effect within, you know, however far of the, of the, of the root zone.

Well, that, that's too bad, Ryan, you know, because I really like the idea of us being able to, to make feminized seeds in a more, you know, natural way than using the chemical. Right. I really liked that idea and I'm disappointed.

We can do that. Right. And the way that we do that is, is like I said, the way those early Dutch seed producers, the rotalization style. Yeah, yeah. And those, you know what, there's hemp lines that also produce that. There's a, There's a hemp breeder who's now passed, but it was a Hungarian hemp breeder named Ivan Boshka, and he came up with what he called a unisex line that produced nearly all female Plants, we're talking, you know, 990 out of a thousand kind of thing. And he had this method for going and selecting intersex plants, but only the very right intersex plants. Like he had a subset of plants that, that he chose. And when you use those plants to breed with regular female plants, they were, they were, you know, air quote, gynoecious seeds are all female seeds. They weren't all female seeds. They were feminized seeds with a small percentage of intersex plants. So you can do that stuff naturally. You know, I don't think that we need to be scared of chemicals like silver nitrate. I mean we, we feed all sorts of nitrates to our plants as food. It's just silver and nitrogen and oxygen. And I think if you're careful with it and you don't spray it on yourself, there's really very, very, very minimal risk associated with doing that. And I think it's more of a lifestyle choice to say, hey, we're not going to use chemicals like that. But I think that the trade off, if we're truly looking for plants that are sexually stable and aren't going to throw intersex plants, we're better to rely on the chemicals rather than going and selecting for plants that are likely to produce pollen under any given number of triggers, be it low light conditions or like we said before, ph or certain feeding regimens. Right. So I would not do it that way.

Right on. I want to push one more on the, on the silver thiosulfate. Is this, does this occur in nature? Like is it a man made chemical or are we just taking something in nature and applying it to the plant? What I'm getting at is I'm trying to understand for myself. I mean, certainly it's not something as far out as genetically modified organisms. We're not doing genetic line changes or anything. We're pushing the plant to do something with a chemical and many chemicals are natural. I'm trying to decide how unnatural, if you will, using STS is. I mean, is this bastardization of nature or is this just a cool natural trick?

I don't think it's a bastardization of nature. But also keep in mind that I'm, you know, I am pro responsible use of genetic modifications because you know, technologies, you've heard me say this before at the Emerald cup, but you know, technologies like Talin and Crispr are going to change. They are changing the face of plant breeding and allowing us to do things that we could never have done in the past. You know, in terms of our gynoecious seeds. Natural. Absolutely not. You know, there's no all female seeds in nature. There's, there's monoecious seeds which are, I think it's important to say again here, monoecious seeds. They're really all female seeds but that have an intersex trait. So if you look at them genetically, they don't have Y chromosomes, but they're heavily intersex plants. Right. So technically, you know, on a genetic level there they are female plants are all female plants, but they're all female and intersex, which makes them phenotypically monoecious. Right. So yeah, if you want to be hippie minded and only doing things that are found without the help of man, feminized seeds are not for you.

Right on. That is very interesting and I'm super grateful to Kelly for, you know, suggesting this possibility in the tent so that I could go down this path. I mean, hell, here we are, we're like a year later and I'm still, I'm still talking about it because it's such an interesting idea. All right, well, we've had a super long first set. Let's go ahead and take our first short break and be right back. You're listening to Shaping Fire and my guest today is cannabis biologist Ryan Lee. While I love growing under the sun, there's a lot of good reasons to grow indoors. And if you're like most folks, you want a lighting source that grows high yielding, healthy plants without using excessive amounts of light electricity. BIOS lighting creates biological lighting solutions that brings the natural brilliance of the outdoors into your grow room. BIOS lighting has the attributes that I look for in a horticultural lighting solution. 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So in the second set, we're going to talk a lot more about feminized seed myths and figure out which ones might hold some water and which ones are just are just holdovers from the early days of feminized seeds when they were made with rotalization. So, Ryan, we talked a little bit already about how poorly tested seeds, poorly tested feminized seeds can go intersex when put under stresses and really high end. I'm not even really high end. Properly made feminized seeds don't do that. And so I guess my question for you is if the goal when making feminized seeds is that there are not going to be males and that they will not be throwing male botanical parts, what can we do when producing feminized seeds? So that doesn't happen because, you know, we have to cop to the fact that sometimes that does happen. I mean, we see the pictures on Instagram. Instagram. But the idea to say, yeah, that was it, that's an untested plant. That's a good position to take. But then what's that testing look like? So that you're not making feminized seeds that crack under stress.

Yeah. So I mean, there's all these conditions that under which even regular cannabis females that are created through regular matings with males and females, they also throw intersect what we call intersex characteristics or, you know, as they're Colloquial, colloquially. Sorry, colloquially known as hermes, which is really not a very good description for them. But these intersex plants, they pop up occasionally. It's just a factor of cannabis. It's part of the cannabis genome and it's present to a degree or less in different cultivars in different varieties. So typically what you do is you expect a plant is going to undergo this stress phenotype of intersexuality. You expose it to those conditions that might trigger that intersexuality to come out. So if you flick the lights in the middle of the night and give them periods, essentially interrupt their dark periods, that can sometimes cause stress. And in some plants will become intersex and some plants won't. Well, the ones that become intersex, they're probably not good breeding candidates because they're going to be triggered by, you know, light interruption. The plants that are resistant to becoming intersex, in those, in that condition, they might be a better candidate to be part of a female, Female. Female breeding program. And so same goes for, you know, temperature fluctuations or ph fluctuations in the root zone and a lot of these stresses that might trigger. Again, we look for the plants that are triggered and we select against those plants using the principle of like begets, like, you know, anything that has that trigger, we want to remove those plants from our breeding population so that those traits aren't passed on. And I think that any, really, any plant that is made to be part of a female breeding program or a self, that those seeds should be examined before they're sold. You know, they should be grown out by the breeder and confirmed that they don't have intersexuality. That's one of the things that we do when we, especially when we use self seeds is you're not just giving, you know, when you use a regular breeding program and you take a plant that might have an iffy characteristic, it's 50, 50 whether that characteristic is going to get passed on. Because it's, you know, there's, there's pollen, there's genetic contributions coming from another individual, which is also going to influence the progeny. But when you're selfing seeds or you're selfing plants, those resulting seed lines, they kind of get a double dose of whatever good or whatever bad traits the plant might hold. And so we really have to go through and do a subsequent screen and make sure that that plant isn't passing on bad characteristics.

When you're doing your breeding programs, what do you choose to do to the Plants to stress them out. Like, you know, it's funny to hear people joke on Instagram about the crazy obstacles and torture they put their plants under to try to weed out the strongest plants. But so far you've mentioned interruption of light cycle and temperature swings in the root zone. So other than that, what other, you know, trials would you put plants through to make sure that when they're under stress, they're not going to go intersex?

Well, I mean, just normal grow room stresses. The problem is, is that, you know, I try to, I try to create an environment that the plants are going to grow in for the breeding. You really want to select the plants under the same environments that they're going to be growing in. And so, yeah, you can torture plants. And I expect that there's a certain degree of torture that plants do go through being grown in home gardens by inexperienced growers in little tents and stuff. You can't, I mean, I guess you can try to build a bulletproof plant, but really I just do selections in greenhouses and expose them to the normal everyday stresses that they might get. So it's kind of a, it's kind of a. I don't really, don't torture test too much. I mean, our torture test is using the plants in production. Typically, I don't use plants unless it's for a very small breeding program that doesn't become a commercial release. You know, something to do with like an obscure chemistry like thcv, for example. Really, a lot of the breeding becomes focused on plants that are accepted by the marketplace. Right. So you want to take the blue dreams and the cookies of the world and the OGs and make them better. Right, Because I think that all those plants, we go through this kind of fashion thing with cannabis. One plant that's fashionable one day, seven years down the road, nobody wants to smoke it. But, you know, if you wait another seven years, it's like bell bottoms, they'll come back.

Well, in addition to the, you know, the belief that feminized seeds are more likely to go intersex, another belief of folks is that that feminized seeds are less vigorous or produce plants that are smaller in size. And, you know, I have produced smaller plants from feminized seeds. I've also produced monster plants from femini seeds. And I've kind of put that more on me and my inconsistent growing. So from the scientific background, are feminized seeds less vigorous or smaller in size than a true reg seed?

Well, no, they don't have to be because again, you could make an F1 hybrid feminized seed line by taking two stable true breeding females and crossing them together. Right. And so the plant would be that, that plant would be an F1 hybrid. So that's, you know, those are typically very high, very high in vigor. Southing though, again it, it we get a double dose of any traits. Any, any, you have an opportunity to get a double dose of any traits that are in the parent plant. And so any plants that are for diminutive stature, they kind of. Yeah, we do see in cell seeds that you have this wide range of, of plants that, you know, some that are very vigorous, some that might not be so vigorous, some that are very small with tiny leaves and some that are normal looking. And that's part of the breeders goal is to select for or against those traits depending on what the end goal is.

But that has nothing to do with the actual fact that they're female seeds. That's just like straight up breeding.

Yeah. And I think that that's really maybe one of the most important take homes that we can, that people can have coming out of this conversation is that, you know, what happened. So when people started selfing seeds, they would take these plants and they'd spray them with STS and then they would grow the resulting S1 seeds. And maybe 50% of the plants were small or 50% of the plants were intersex like we were saying before, you know, and the assumption is, oh well, I use this new breeding technique that's probably what caused the problem. Whereas the truth is is that the breeding technique exposed a genetic weakness that was unseen in the plant. Right. And we have to be able to tease apart the cause of the, the cause and effect of the consequences that we're seeing. Right. Like was it really the sts that we sprayed on the plant that made the plants in the next generation subpar? Or maybe it was that those plants that were generated, they just inherited some subpar genetics that were buried in the plant, which is not only possible, that's in fact what we expect from selfing outcrossing species like cannabis.

And in some ways it's, it can be seen as an attractive attribute because if you are a serious breeder and you really want to pull all of these less than attractive attributes out. Well, you know, creating feminized seeds and being able to see all of the stuff, that's actually a win if you don't want to put out stuff with this, these attributes hidden.

Yeah, it might be a condition that, you know, for example, you know, there's some plants that if you cross a white flower with a. With a red flower, you get a pink flower. But when you try to breed that pink flower, the next generation produces 25% white flowers, 25% red flowers, and 50% pink flowers. Right. So it's like the. If you want to have all of the flowers come out pink, you have to cross the red and the white types together to get pink. Well, you might not like the red and the white types, right, but they become a means to an end. For example, a lot of these traits that we see as favorable in elite cannabis plants are as a result of what we call heterosis, which is a condition where the plant has two separate chromosomes from diverse and disparate parents that are very genetically unrelated. And so you kind of have to go to the ugly before you can breed the beautiful. Right. It's like one is a means to the other, and they kind of exist in a balance. You can't have one without the other.

That's like dying to be a meme, dude. You have to go with the ugly before you can breed the beautiful.

In a way, it's. That's true. Right? We don't. I don't know. It's. I don't know. We. There's all these cross analogies that you could use, but when you start talking about breeding humans, people get upset about it. So.

Yeah, well, let's just. Let's move right on from there. But I do want to hit on something before we go to the next potential myth to bust. You know, you came up to the point where you're talking about, like, you know, you take the pink flowered plant, and then when you breed that, that only 25% of them or some percentage are going to be the pink, and then maybe the rest are going to be white. I don't know a lot about breeding myself at a fine level, but I have studied a little bit of genetic trait charts and how they're passed on. Do you have a book that you would recommend to people who have not studied this at the university level who are just regular cannabis folk like I am? That would be a good read. That would would explain, like, the next level of cannabis breeding, but without burying me in scientific, you know, jargon that I may not have the education to, you know, follow?

Yeah, I mean, it's an old one and it's not very advanced techniques. It doesn't really discuss self ring or anything like that. But Rob Clark's book on marijuana botany, the propagation of distinctive cannabis. Marijuana botany is just an incredible book, it's a classic.

You know, even though some parts of it are a little dated, it's still like worth every, for everybody to read.

Yeah, that one, the only other one from the cannabis industry that I would really recommend is as Jorge Cervantes, Marijuana Horticulture. It was a green book and I think they called it the, the Medical Growers Bible or something like that. I actually wrote the breeding chapter for that back in 2004 or 2005 and I tried to lay it out in a very simple, goal oriented way of breeding that I had hoped that it was easy for the consumer to understand. And it was a little different than most. Most breeding segments in these cannabis books had been presented in the past. So that would be one worth checking out. And then as far as anything else and really trying to understand the meiosis and the genetic dance that goes on behind, that's really the foundation of all plant breeding. Plant breeding for dummies or Genetics for dummies, something like that would be really great. And they're, you know, 10, 15 bucks online. I'm sure people could find it.

Yeah, they give you a nice broad base. It's not going to be cannabis specific, but you're going to learn a lot from it. Yeah, that makes sense.

I think that you'd learn more than you would from learning, learning from a cannabis book, to be perfectly honest. More principles that really lay the foundation of the science of like you were talking about punnett squares and you know, all that kind of stuff. It really gives you a visualization.

Yeah. Actually maybe read the Breeding for Dummies version of the book and then go read Rob's book. So you already have a basket of knowledge with which to read Rob's book. I was very early in my cannabis life when I read Rob's book and it was like over my head. I would probably actually do way better if I read it now.

Yeah, if you went back and you read it now, you'd get a lot out of it. Everybody has to remember too, that book was written almost 40 years ago now. It's 1981, it was published. So it's not going to be all full of stories of OG and Blue Dream or cookies, but it does really a beautiful job of talking about all the different land race varieties and it gets into some basic trait breeding.

It just crossed my mind, I'm like 81. I mean, wow, that knowledge is as old as when I was listening to Van Halen. That's a long ass time ago.

So.

So the next belief about femme seeds I want to hit is, you know, a lot of people will say that feminized seeds may be great to grow for your own pleasure, for your smoke, or for your medicine. And it's certainly great for patients because they don't have to deal with, you know, male plants, because there's nothing quite as sad as a patient that's growing, you know, six plants for, you know, themselves, medical, and then they end up with four males. Ugh, that's so sad, because not only are patients usually really emotionally attached to each plant, but that really decreases the amount of medicine you've got for your year. So that's a drag. But. But a lot of people say that using feminized seeds to breed is a bad idea. Or so since we've been talking a lot about cookies, let's say, like, okay, you know, you get some girl Scout cookies, feminized seeds. Somebody would say, yeah, if you want to breed with cookies, you need to go and pheno hunt, you know, some cookies seeds that are regular. Or get yourself a regular clone and breed with that. That you don't want to breed with the feminized seed because there is something at its very basis that is, you know, more easily corruptible than using regular seeds. What are your thoughts?

Yeah, I think that that's nonsense. I think that, you know, we try to look at it scientifically again. If you look back, there's that for me, if a plant is good enough to use as a female breeder, like, if a plant is good enough for me as a breeder to say, okay, this plant is uniform, it's stable, it doesn't have intersex offspring, it doesn't go intersex when the lights get flicked on at night or the pH of the soil is too low. I'm confident that this plant is a reliable, solid breeding candidate. For me, that plant is good enough to either be a pollen donor or a pollen recipient. There's no difference. But I think that a lot of people in the scene believe that the STS creates some kind of chromosomal unstability. In fact, I heard a plant breeder say that at the Emerald cup last year, and there's just no evidence that that happens. I mean, theoretically, I guess it's possible that somehow the silver damages the DNA, but there's never been any kind of scientific confirmation that that happens. And we got to remember that these. These techniques are used widely in other crops. All your cucumbers are pretty much feminized or gynoecious lots as well, because it's not efficient for farmers to be growing acres and acres of plants when 25% of the plants are male and they're not going to produce cucumbers, that's a huge waste of time and space. If you're a grower. If you're a grower, you want them to be. You want to fill your plot with everything that you need. I don't think that we've, you know, I don't think that people have been able to tease apart again, it's like we said before, how much of the damage, how much of the problem are we seeing is a result of a bad selection of a parental plant versus the breeding technique that was used to create that, the generation. Right.

Yeah.

And for me, I just don't believe I haven't seen enough detriment across the board to say, yes, I think that this is a result of the technique and not the plants that were chosen. I think that you can attribute all the negative consequences of selfing from genetic weaknesses that are contained within the plants themselves that were selfed.

Right on. All right, so last question before we go to our second break. You know, I know this is going to be kind of a vague question, so I'm going to need you to work with me here. But I want to ask you it, because I see it so commonly just kind of thrown out there on cannabis forums by people who are hating on feminized seeds and they will say some version of you shouldn't use feminized seeds because growing or breeding with feminized seeds limits the size of the overall cannabis gene pool and it's bad for the future of cannabis as a whole.

Yeah, it's a funny statement. And there is a, there is a bit of a truth to it. I wouldn't say that there's truth with it pertaining to feminized seeds. Everybody has to remember any seed or any child can only have two parents, right? So the, the act of taking two plants and putting them together, or taking a thousand males or a thousand females and putting them together, they both result in the same consequence, that every seed came from two parents. Right. So if you're taking two female plants and mating them together, or you're taking one female plant and one male plant and mating them together, you're still doing what I call a one to one mating. Right? And so, yes, one to one matings create a genetic bottleneck. You're narrowing the genetic possibilities that are being passed on. Sometimes as a plant breeder, you need to do that. Right. Because you do need to purge when you're trying to create something new, you have to remove all the traits that you don't want. And sometimes the way you do that is by creating a tight bottleneck. You can always take that bottleneck population and backcross it to the more diverse family. But I think trying to assign the blame of genetic bottlenecking to feminized seeds really isn't accurate because that's what plant breeders, cannabis plant breeders, have been doing since dawn immemorial.

I mean, essentially that's the goal.

Right.

If we're trying to create consistency in our seeds so that when you plant the seed, you get the plant you're ideally looking for, that's actually what happens with then regular seeds as well. It's a honing process. It's like sculpture. It's removing the parts you don't want. Now, if everybody did that with all of cannabis at the same time, well, all right, that may limit the gene pool theoretically, but since that's not actually what happens and we can always out cross the realistic experience we have is not actually a decrease in the gene pool.

Yeah, I just, I don't think, I mean, I do think that we do need to be careful with genetic preservation. All crops are going through this massive narrowing, but I think attributing that problem solely to something with feminized seeds is a bit of a false dichotomy. Right on.

Fair enough. All right, so let's go ahead and take our last short break and be right back. You are listening to Shaping Fire. My guest today is cannabis biologist Ryan Lee. In times like these, when so many cannabis companies are growing their flower in gigantic warehouses and fields using synthetic nutrients, it is good to know that there are authentic California heritage growers using natural farming techniques and sunshine to cultivate cannabis flowers for you. California has produced the best cannabis in the world for generations. And the idea that massively scaled industrial cannabis production could produce the same quality as small batch, lovingly cultivated flowers is just silly. Moontime Medicinals is located in Humboldt county on the lush south fork of the Eel river watershed in the epicenter of the American cannabis heartland. 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I'm your host Shango Loess and our guest this week is cannabis biologist Ryan Lee. So Ryan, in this third set I am hoping that you can take us all down this path of female only breeding. And honestly I do not know a lot about this. I only have two friends that practice female only breeding. But my God, are they like excited about it and they really find it as like kind of like being way more immediate and successful more quickly and I don't really understand why. So I figure since we're doing a show all about female seeds and the producing of them, the breeding with them, let's go ahead and talk about female breeding and kind of, you know, bring everybody up to the state of the art. So why don't you start generally with just what is female only breeding.

So really it's exactly what it sounds like. It's breeding without using any male plants. And and it's a breeding tool or technique that is really incredibly well suited to breeding specifically for drug cannabis when you're breeding, because drug cannabis is the experience is so based on the chemistry of the plant. Really what we're doing is trying to create a special chemistry, both in the cannabinoids and the terpenes and terpenoids and all these other chemicals that the plant produces to create a user experience or a plant with a user experience that we find enjoyable. And when we use traditional breeding techniques, where we use a male cannabis plant and a female cannabis plant, it's really easy to choose the female cannabis plant, because we can use a laboratory or we can use our nose or the way that the smoked flowers make us feel as a guide to which individual female plant is the best or better than all the others. Where we can't do that with male plants because you don't smoke the female. You don't smoke the flowers of the male plants. The smart. The male plants really don't produce a smokeable flower. And because they don't produce a flower, we can't. Or they're, sorry, a flower that produces a lot of resin glands. We can't really use the same laboratory techniques that we use in the laboratory. I mean, I suppose we could, but it would only give you a little bit of a hint as what the plant would look like if it were a female plant. Whereas if we choose female plants based on their bud and their flower characteristics, we can kind of do what I call positive selection on both sides of the cross. We can find a pollen donor plant that has traits of interest, and we can have a seed mother plant that has maybe the same traits of interest or different traits of interest. And that way we're doing positive selection on both sides of the cross. Whereas if we were trying to breed a male with a female, we could, you know, we could understand everything that that plant genetically contributes to the female flower on the female side. But on the male side, we're kind of shooting in the dark.

So. So what. What I'm. What I'm picturing in my head is I'm going to take one female plant, spray her with sts to cause her to give off male pollen, and then I'm going to pollinate that to a second female. And so both of these female plants have got traits that I'm interested in. And so hopefully the seeds from that second female plant, when I grow them, will carry the phenotypes and the traits that I want from both plants, and then I will secondarily choose from those.

Yeah. More or less. Yeah.

All right. So if it's more or less, clean it up.

Right.

Because people are listening and they're gonna want to do this.

Yeah. So if you're listening at home, I always Recommend that we treat our donor plants as if they're on about a three to four week schedule ahead of your, your female target plant. So you always want to take your donor plant and trigger it into 1212 or trigger into flower by putting it in darkness or you know, 12 hours of darkness. And on the first day of doing that, you spray the plant with STS, which becomes an application that I usually do weekly until their third or fourth week. We then wait two to three weeks for our target plants until we move them into the flowering chamber. Because the sds, it does slow the plant down a little bit and I'm not sure why it does that, but there seems to be a delay or there's no synchronicity in their flowering periods if you don't trigger the pollen plants before. But yeah, it's really about, it's really about selecting plants on both sides that you find interesting and bringing them together through this STS process. And there's a lot of benefits to it. One, as you said, Shango, you're more quickly able to move towards your breeding goal because again, you're doing positive selection on both parents, not just the one, but secondly, every time you want to grow out those populations that you're experimenting with, your test crosses, if you're using male, female breeding, 50% of those plants that you have to spend time, energy, nutrients, plants, water, soil, taking care of and bringing them to maturity only to find that they're males is really, it's kind of a drag. In fact, it's a real pain in the ass if you're trying to do it on any type of scale. Whereas when you're using all female breeding, again, one of the benefits is that you're receiving gynosha seeds out of the end. And therefore every seed that you plant is going to be a potential breeding candidate or a winner. You know, a plant that you can look at, it's not just taking up space until you feel, you find out that it's a male. So that's one real benefit. But I think the benefit of being able to really look at the chemistry of both plants before you bring them together is a huge boon. And you know, in our work at Napro, we really, we showed that that was a really fast way to make progress when you're breeding for specific chemical types.

Even though I do like the process of choosing male plants compared to actually using a female plant that is directly displaying the traits that I want, it makes talking about tight internodal spacing and rubbing the stem of a male of a male plant to guess what the terpene profile would be like. It makes it seem like caveman, like pure prehistoric.

Well, it really is. It is really caveman style. And you know, when I, when I do that type of breeding, and it's not to say that I never use males in breeding projects, but typically when I do use males, I won't use a single male. I'll collect pollen from, you know, 10 to 15 plants or maybe more. But you only select, say 10 to 15% of the, the plants that you're growing. And you're really looking at a whole set of characteristics, you know, resistance to molds and mildews and that kind of stuff. And all the, you know, like you said, the leaf shape and the inner node distance and the floral structure. You can kind of get an idea of what you're looking at and use that as a loose criteria to select away 80% of the plants. But I still use the remaining 20%. So I'll collect them all and mix all their pollen together because like we were talking about earlier, that provides some level of genetic diversity. But if we're really trying to get, you know, tight down and dirty with one specific chemo type or one specific smell profile, it's a lot easier to take two plants that already have that smell profile, you know, either as determined by the lab or by your nose, and using those plants to mate together. And you'll see that the end results are vastly different. So you much more quickly approach your goal. If you're using females on both sides.

I can totally see how positive selection, right, Choosing the traits I want in the two females to get there more efficiently. That makes total sense to me. But I know that there's a whole bunch of folks out there who are going, oh, no, man, like, I absolutely need to use a male or it's not, just not, right? So if we were to science that up a little bit, is there anything lost by not using a male?

There is. There's a Y chromosome that's lost, and it's not lost. I mean, we can preserve Y chromosomes in a handful of seeds in a fridge for, you know, 20, 30 years. So it's not like we're in any fear of actually losing the existence of the Y chromosome. But the truth is, even when we're growing those regular seed lots, or what we call regular seed, lots of those male plants don't fall into our marijuana production, right? We don't into the production of cannabis flowers. We just don't use them. Males are essentially irrelevant to that process. Otherwise, you know, cannabis has, in its genome, cannabis has 10 chromosome pairs, nine pairs of autosomes and one pair of sex chromosomes. So when we're using, you know, when we're really, what we're doing when we're breeding is we're just shuffling copies of those 10 chromosomes. But when we, again, when we look at marijuana production, we're only concerned with the variation present on the nine chromosomes and what's on the X chromosome, because what's on the Y chromosome is really kind of irrelevant for drug cannabis production. There is something called crossing over in which Y chromosomes and X chromosomes exchange these little pieces of DNA. And there's something on the male chromosome called the pseudo autosomal region. And all that means is that there's part of the Y chromosome that actually does exchange genetic information with the Y or with the X chromosome. But I don't think that we're truly eliminating anything of value to drug cannabis growers by creating feminized seeds without Y chromosomes.

Right on. Fair enough. All right, so I've got one more. I'm going to ask you a question about sourcing sts. But before we move on off of this, I'm going to, I guess we'll call this another fishing expedition. You know, when I have people on this show that have got a level of technical knowledge that I may not know, I like to ask this question, which is, what am I not asking you about female only breeding that people want to know. Right. You know, I really only, you know, I'm learning about it as you're telling me about it. And so, but you, you are very experienced with it. And you know, what tends to get people excited. So what else is there? What is another female only breeding data point or something that you think is important and relevant that people would want to know?

I don't know. I think that we've touched on most of them. I think the important things to realize is that the two real take homes that I really want people to think about are what blame are we putting on the process of sts that really should be seen as blame of bad selection of plants or plants that had negative traits. And I think the community really assumes that the process of creating feminized seeds does something to the genetics of the plant. And that really hasn't been shown in any type of research anywhere. I think, you know, I think that as breeders, we need to do a better job of testing the seeds and being open about the techniques that were used to produce the products that people are buying and give the consumers as much information as possible. But we also have to educate the consumers so that they know how to deal with that information. You know, it's, you know, it's enough to say, hey, we have these are S1 seeds. But if you don't know what an S1 seed is, I don't think that that information is really too, too helpful.

Right on. So, so, so, you know, there's chances are that there's a lot of people who are going to be all hot to get some silver thio sulfate now. And for me, if I were to do a breeding program like this, I'd probably just call up Matt Riot and buy a bottle of his feminizing spray, which is sts. But I don't really know a lot about this chemical is there. Like, how do we know about the veracity of the STs that we're using? Do they sell it at scientific supply, or do we need to find somebody in the cannabis scene that has made a batch? What's the nature of this chemical and getting it?

Yeah, so typically when you're buying it, you can buy. So if you want to mix your own, there's recipes for it online and there's all sites or all sorts of different types of recipes for it. There's typically two components that you need. Silver nitrate, which is a crystalline chemical you can buy for about, I don't know, it's about 20, $25 for a gram. And then there's sodium thiosulfate, which comes in two forms, which complicates the recipe because there's sodium thiosulfate pentahydrate, which essentially has water molecules built into the crystal and then you can get it without. And the molecular weights are slightly different, so you have to know which one you're working with. If you're trying to mix your own, typically what is done is you make, you measure out a given weight of each of the crystallized substances and you mix them into a set amount of water. And now you have two stock solution which you then mix together and you have to pour one of them into the other and not the other way around. Otherwise they react in a very negative way. Not a negative or dangerous way, but the compounds don't come out as needed. That all said, for most people that aren't playing around with it, you're probably better off to just order it online. I noticed there's a couple of outfits now selling spray bottles on it on Amazon.com and if you're in the United States, there's a company called Hybratech who's a plant breeder friend of ours from Oregon that is, likes to be credited as the first person that kind of created the swell of feminized seeds. Even though he wasn't really the first person to create the product, he did independently develop it and he's doing great work too.

He came up during that same show I referred to earlier with Caleb from CSI Humboldt. They send best practices back and forth.

Yeah, Hybe is, we've always known him as Hybe, but he's a really, he's an educated plant breeder. He really knows what he's talking about and he thinks about plant breeding as a plant breeder would not as of someone that is trying to make money off of seeds. So anyway, the bottom line is that he sells it and it's, it's, you know, it's probably more expensive than you need to pay for it, but it's a ready to use formula that you can just order and it comes in the mail and a little spray bottle and you can just spray that on your plants. And you know, I think if you don't know how to mix lab chemicals, it's probably that that type of product is right up your alley because it is. STS is an oxidizer and, and you know, it's, you don't want to be breaking bottles and putting your hands in it or any of that kind of stuff. It stains really badly. If you, if you, you know, knock it over on a wood table or a white table, for example, it's going to get, it's going to stain it.

For the, for the especially nerdy folks who want to make their own. You mentioned that there were different types of recipes out there. Do you have a, you know, a keyword that you want to drop so that people could use to find a recipe that you find is reliable?

If you type sodium thiosulfate cannabis into Google, you're going to find a whole bunch of recipes. I would mix it a little stronger than most people are making it. Just concentrate it a little bit more. Don't put as much water in. Obviously use gloves, long sleeves, you know, goggles, all that kind of stuff when you're spraying it. Just use common sense. And yeah, you'll be able to find the components online, usually from. Sometimes they sell them at photo formularies, like places for developing film, like old camera film. Anybody remembers what that is?

So earlier when you were talking about the breeding aspect, you said, well, you know, you set the flower or you set the plant to 1212, it starts to flower. And from, from the day that you flip to 1212, you start spraying the plant once a week for the first five or six weeks. Did I repeat that correctly?

Four to five weeks. Usually by that point in time, it's dropping pollen or it's, or the male flowers are well towards development. The key there is that you want to, you want to have. So I call, I have two groups, what we call the target plants and the donor plants. The donor plants are the plants that get treated. They're the ones that are going to become the pollen donors. And so we put the pollen donors into flower two to three weeks before we put the target plants into flower. And that just gives those plants enough time to get well into their male flower development period. So that by the time that they're ready to open and release that pollen, your female plants are then ready to receive pollen or your target plants are ready to see pollen. So that we leave like a two to three week window in between, you know, when we induce the donor plants to flower and then when we induce the target plants to flower.

How much STs do we spray on? I'm one of those people who tends to think more is better. And since I've gotten involved in learning Korean natural farming, I'm teaching myself to not think more is better all the time. But with this. Can you just give us some kind of rule of thumb of how much to spray? We probably don't want it damp, but we also probably need it uniform.

Yeah, it's, it's a little more coloring with client crayons than doing intricate science. You can just kind of spray it. I wouldn't necessarily encourage people to spray it onto the actual leaves of the plant. The sun leaves are, you know, you could, they will absorb the chemical, but it's this, this chemical can be a little bit phototoxic, so when they get exposed to subsequent life, it can damage them. We want to spray what I call the crotches of the plant. So the junction where the leaf petiole or the stem of the leaf meets the branch meets the, either the branch or the main stalk. And that area is where the new flowers are going to grow from. So essentially anywhere that you think you would see, see a bud site on a female plant, spray that bud site or the tip of the, of the shoot at that bud site. But don't necessarily worry about the sun leaf itself. It's more about spraying the ethylene blocker on the area where those flowers are going to produce rather than on the leaf.

Well, I'm glad We hit that because in my head I was actually picturing a pre existing flower. But that's right. If you're going to start this at 1212, you're not actually spraying it on flowers, you are spraying it on future sites. So you're really spray. You're actually probably trying to spray branches at the, at bud sites, but before they're there. So. So it takes a little bit of experience to know where those are going to be.

Yeah, a little bit. And it becomes obvious, you know, after you, after you plant or after you. The plants have been in flowering for about a week or 10 days, you're going to start to see little tufts of flowers. And at that point in time, yeah, you switch to spraying the flowers themselves because we want the. The whole point of the action is for the ethylene to act at that flower site and we want to shut down the production of female flowers and in turn have the plant produce male flowers. So the plant will still grow little buds, but the idea is that those little buds are now made up of male flowers and not female clusters or colas. Right? Yeah.

Right on. Cool. Well, Ryan, thank you so much. You know, you know, I love cannabis in general, but as somebody who talks about and helps patients source female seeds and as somebody who grows feminized seeds, it's always been kind of a mystery how these, how these plants or how these seeds are produced. And I just kind of went on faith. Right. But I feel a lot more, I don't know, empowered, understanding what's going on behind the secret sauce. And also to know that it's not feminized seeds that are often bad. It's actually poor growing, which actually makes, or, excuse me, poor selection, which makes a lot more sense to me. So thanks for sharing your valuable time and your experience and breaking it down in such detail for all of us.

Yeah, it was absolutely my pleasure. Anytime, Shango.

Right on, brother. So if you want to find out more about Ryan Lee and his breeding work, you can go to his new website, which, you know, if you're hearing this the week that it comes out, it's probably. There's probably just a placeholder there for now, but that's going to be@chemavar.com that's C H E M O V A R Keemavar.com is going to be the future home of his company website. And if you want to send him a message or check out his interactions with other folks in the scene, you can follow his Twitter feed, ChimeraGenetics. And that's C H I M E R A Genetics On Twitter, you can find more episodes of the Shaping Fire podcast and subscribe to the show@shapingfire.com and on Apple itunes, Stitcher, and Google Play. If you enjoyed the show, we'd really appreciate it if you'd leave a positive review of the podcast wherever you download. Your review will help others find the show so they can enjoy it too. On the Shaping Fire website, you can also subscribe to the weekly newsletter for insights into the latest cannabis news and product reviews. On the Shaping Fire website, you will also find transcripts of today's podcast as well. For information on me and where I will be speaking, you can check out shangolose.com does your company want to reach our national audience of cannabis enthusiasts? Email hotspotshapingfire.com to find out how. Thanks for listening to Shaping Fire.

I've been your host, Shango losing.