 Well, thanks everyone for coming and sitting in on this. My name is Chris Scram. I work for South Dakota State University, but I manage a farm, a research farm over on the western side of the state near Rapid City. So seven and a half hours or so from here. I'm an associate professor in the plant science department, and my background is really in more in the soil health and in microbiology. And so I'll get into a little bit about that later on. I was asked to talk about some of the work we're doing with nitrogen fixation. But I thought while we're here, I got a little time and talk about just pea production in general and then get into some of that. And so I'm just going to introduce some of the work that we do in South Dakota. I think, you know, obviously North Dakota is a pretty major player in a lot of the pulse growing and so much bigger than South Dakota is. And so we learn a lot from NDSU and a lot of the growers here as well. But yeah, so I just thought I'd talk a little bit about what we're doing. But at any time, feel free to interrupt me and ask questions. Certainly no problem. I kind of like having that back and forth anyway. It helps, I think, with the discussion rather than just a one way monologue. So this is actually a picture of our farm here on the left. And you can see the background there is bare butte if you're familiar with Western South Dakota. So that's kind of where we are and right to the west of us is the Black Hills. So we're pretty high elevation, much higher than here. A bit shorter growing season but obviously similar climate. So, you know, I think we've talked a lot about this probably throughout the day and a lot about where are these things going. And so the peas in general, there's two market classes. They have greens and yellows. And I think that trips people up sometimes. What's the difference? There's not really much difference in terms of growing them. But they have different end uses sometimes. And so obviously a lot of these are exported. A lot of peas are exported for edible food and then exported for food aid. That picture in the top left there, I was in Ethiopia a couple years ago doing some work and there was this massive warehouse. And it was just stacked with these bags of split yellow peas. And so I thought that was pretty interesting. And so it's a big part of food aid forage obviously. And then this newer market where it's fractioning for protein. If you look around like at Target or, you know, Safeway or something. Whenever you see stuff that says fortified with protein or whatever, it's very often it's peas. Or, you know, there's a few others. There's pea milk now, that picture, that ripple there. I took that in Target. And of course the meat substitutes. And so there's a growing market I guess for this fractionated pea protein. Pea, you know, it's roughly 23 to 27% protein. And so they seem to like it for that. And of course those snack foods and things keep expanding. So this is kind of the, this is the growing area for much of the country. Certainly doesn't stop at the border there and it's pretty significant growing region in Canada. But you can see it's up here in the Northern Plains, Montana, North Dakota. And it's starting to migrate south more and more each year as those markets expand. You know, and for those of you that don't grow peas, if you grow spring wheat, it's very similar in terms of timing and things like that in the growing season. Very much like spring wheat, planting in the early spring. You know, for us it's early April, maybe it's a bit later here. And then it comes out about the same time as spring wheat does in the summer. Seeds will germinate above 40 degrees. You know, so that gives you a sense for when you might be able to start growing them. And they're hypogeal, which means that the growth point stays below the ground during the early part of the growing season. And the value in that is that it can tolerate colder, you know, when you get these cold snaps coming through. You might get some leaf burn on the top, but you know, the growth point stays below ground where it's a little bit more insulated. And so it can tolerate frost pretty well. So like I said, we get a lot of questions about green peas versus yellow peas. And I think by far yellow peas are the dominant pea in the market. And so these are two box plots that I'm showing here. Can you see my mouse here? Not really. Okay, well I'll just have to point to it. So you can see on the right hand side there's the box plot with a line in the middle. That's our average yield for yellow peas over quite a few years. And green peas on the left. And so the thing really to pay attention to is just that thick black line, that's the average. And it's generally a few bushels more for yellow peas than it is for green. And sometimes that's just the result of the varieties that we're testing. But often yellow outperforms green a little bit in our trials. And that's the average protein there too, 26, 27%. It's not much different between the two. So we get a lot of questions as I said. And so that's kind of the standard answer. Yellow peas, traditionally it was a lot for the dog food market. And so the yellow peas always went into the dog food market. And that kind of dominates a lot of the breeding that's going on. These are just some seeding rate studies that we've done. And the top bar that you can see, well, I'll back up. So typically, you know, seeding depth is a big seed. It needs to buy a lot of water to germinate. And so it needs to get down into moisture. So we typically seed them a bit deeper than we inch and a half to two and a half inches. And in the target seeding rate that we've always used is 350,000 seeds per acre. And, you know, 1,600 to 3,000 seeds per pound depends on the variety. So if you do the math there, you're handling a lot of seed at 350,000. It's a bulky product. So the seeding rate trials that I'm showing here in that box plot, basically you can see from left to right on the x-axis you got 250,000 seeds going all the way up to 400,000 seeds. And if you look at those dark lines there, the average is it's not all that different, really. You don't see a big difference. At, you know, there's a slight jump, maybe a couple bushels increase at 300,000. 350,000 are about the same. If you look at the, these are called box and whiskers plots. And so you can see the lines that are coming off of the bars. So that kind of gives you your 75th and 25th percentiles. So basically if you have bigger boxes and bigger lines, that's more, the more spread in the data, it's uglier data. But I also have the points there. So you can see there's a big range within each of those categories. Yeah. Do you find you get more weed suppression with the higher planting rate? So that's, yeah, that was kind of my argument. This is not organic. This is just traditional. So we use, we have herbicides in this. And so my argument for keeping that 350,000 is certainly, the biggest issue that I see with peas is, you know, let's say you try to plant it at 40 degrees. It might sit there. It'll germinate, but it doesn't go fast in the early growing season. And so it might sit there for 12 or 14 days before you start to see any growth. And the, the, the result of that is that it doesn't compete well early on in the growing season with weeds. So, yeah, I think, I'm glad you bring that up. I think that's a really, it's important to have that canopy closure as fast as possible in these things. And the value is that, you know, the peas have the tendrils. And so they'll tend to, they tend to grab onto each other and they form a pretty thick mat. And so you can grab, you can grab some peas and shake them. And you can see a whole, you know, 10 or 15 feet shake with it. And so it does form a really nice canopy. It just takes a long time in the early growing season. So, yeah, so the, you know, the short answer is, yes, I think there's a lot of value in keeping a slightly higher seeding rate. And then protein is there on the bottom. That's the second graph here at those different seeding rates. And that's not really affected either way. And it's not like wheat, you know, wheat has a protein market where you get docked or you get a premium every once in a while. Peas doesn't have that yet. And it would be great to see. I think it would make this a lot more interesting. But at this point, it's just, there is no, there is no premium for growing more protein. Planning date matters a lot. And I know this is context specific, so this is down in South Dakota. But these are two planning dates. One is April 23rd and one is June 3rd. And so that's a month difference. But basically, there's a massive fall off as you start getting, I think our last planning date is about May 5th anyway. So this is just, we were doing some other work and we were looking at this. And there's some varietal differences. These are all, each one, the blue and the orange is the two planning dates for each variety. You can see there at the bottom. And so there is some different varieties that will respond differently. But overall, it was roughly a 65% reduction in yield as you start getting out. Anyone want to guess why that is? What the yield reduction comes from? Yeah, exactly. It's that flowering period. These are really sensitive to heat and flowering. And so when you push that, that planning date out, you start to push it into that early June, later June, when it starts to get 90, 100 degrees. And that really stresses those flowers out and the pollination just doesn't happen. So yeah, so the take home here is early planting is generally beneficial. And this is kind of talking about some of the heat effects on seed set. And so I have a couple of things here. Roughly 40 to 50 days after emergence is when flowering begins. So you can kind of do the math based on when you plant an emergence. So 38 to 45 degrees, it takes about 17 to 21 days to emerge. Whereas at 50 to 55 degrees, it's 10 to 14 days. So you might have a weak difference in emergence. So there's some room to play with there. And this test on this table here on your right hand side, it just shows that once you start to get up above 92 degrees or so at 97, your pollination is really affected. So we have a lot of days where we're sitting out there taking measurements and it's 98 degrees and you can just see those flowers wilting. So it's really critical to try to get those in, like I said, early as possible. I will say though that there's a big difference in these tests. So at 91, it wasn't much affected, but once much above 95 degrees, that's where it really starts to affect the plants. So we do a lot of variety trials similar to NDSU. We stick to the western, you know, similar I think, in a lot of ways to North Dakota to the east of the Missouri River for us is it's more or less Iowa in terms of crop rotations. It's corn and soybeans. To the west, we're a lot more diversified and it's a lot more cowboys first and farmers by necessity. But so the growing regions for pulses are generally central and west for us. So recently, this is just some of the trials from this past year. We do 40 to 50 varieties of tested every year. And the companies will send this stuff in. They'll just send in whatever they think might be a value or might make it in our area. And so it was a rough year, but it was also a strange year for us. Sturgis is all the way out to the west. We had 13 bushels average. And then wall, which is the one in the middle there was hailed out. And then the one in the center part of the state near Peer had 40 bushels. And so there's this huge gradient in terms of weather and drought. And so just highlights the value of having multiple sites. Otherwise we lose a whole year of data with this weather. So this is just some of our products, you know, we put out every year we put out these these tests. And so these are all the varieties that we tested. And we generally highlight the ones that are doing better. And we'll put like a three year average to your average, you know, NDSU does a very similar thing. And so we have protein as well. So now I'm going to transition a little bit and talk about why I was asked to come here was more about the nitrogen fixation process. And so this is a lot of work that that we do. And ultimately what we're trying to do is develop new rhizobia species that we can use for inoculants. And so how many of you are familiar with the nitrogen fixation process? I don't know how long it's been on the slide. Okay, most people are. So I'll move a little bit quickly on this slide. This is basically explaining what the fixation process and we never heard of it. I think it's not a very intuitive term. And so I wish we could we could come up with better terminology. But anyway, the irony of the whole situation is that the atmosphere is made up mostly of nitrogen. And the irony is that you can't plants can't use it. It's not in the form that plants can use. It's a triple bonded nitrogen, very stable. And it's hard to break that triple bond. But legumes, peas, lentils, chickpeas, soybeans have developed this amazing process that they can form this symbiosis with the rhizobia, a certain type of microbe, which can, these microbes can fix nitrogen. They've evolved the ability to do that. So basically what they do is they take the nitrogen out of the atmosphere, break that double bond, attach some hydrogens to it and make it available to the plant in the form that's available. And it's not a free process. So the plant essentially nodulates, meaning they send out these signals, say, hey, we're open for business, certain rhizobia will respond. And so it's very specific to the plant. So like a soybean and a pea will send out different signals and they'll attract different rhizobia, meaning that you can't use the same, you can't use the same inoculant that you would for soybeans as you would for peas. They have different species. And so you look at the roots and you see these nodules and inside of the nodules is where these things are fixing nitrogen. And in exchange for the nitrogen that the rhizobia send up to the plant, the plant gives the rhizobia carbon energy that it fixes from photosynthesis. And so it's an expensive process for the plant. It's roughly 30% of all photosynthesis goes to the rhizobia. So it's energetically, it's a very expensive process. And so ideally, though, it's a mutual benefit. And so that's what we're going to talk about in some of our work in trying to supercharge these rhizobia. So it's hard to overstate, I think, that the nitrogen fixation process and how amazing it is in my opinion. And so what we do in industrial processes is the Haber-Bosch process. So these guys won a Nobel Prize for developing, and it's essentially nitrogen fixation, but it's at an industrial level. They do it and they form urea. And so I wanted to just, I just wanted to, I tried to break this down and see how much energy it actually takes to do this process. And so it takes 18 gigajoules of energy to produce a ton of nitrogen. So one ton of this, of what eventually becomes the fertilizers that we're pouring out on the ground. Globally 160 million tons of nitrogen is produced by Haber-Bosch every year. And so for every ton of nitrogen produced, the energy required, and this is usually from natural gas, but the energy required could power an average home for almost half a year. And so I was just doing the math this morning, and the total energy used to make nitrogen globally could power 80 million homes for the entire year. And so this is this massively energy intensive process. They use some metals to make it happen. And then it's also, it's extremely, there's also a high CO2 emission from that as well. So it just, the part about making or powering 80 million homes from all of our fertilizer use, it's interesting that these little bugs in the ground have evolved this process to do it inside of a plant nodule. So, let me see what time we have here. This has done it at four, okay. Good, we're good. We're good, okay. So as I said, legumes have evolved to take most of their energy or their nitrogen needs from the atmosphere eventually. On average, it's about 40 to 70% of the nitrogen in the plant is from that synthesis symbiosis process, but there's a huge range. And so really anything that affects photosynthesis affects how much nitrogen is fixed every year. And there's roughly five to 20 genes responsible for the fixation process. And so it's really hard, it's hard to manipulate the fixation process because if you manipulate just one gene or maybe a couple genes, it's hard to optimize that because there's a lot of different stuff that's being upregulated and downregulated at the same time. And so from a breeding aspect, it's difficult to do this. And so that's why we focused on the rhizobia instead. Typically, you know, for most of you that grow, you've seen these, there's different inoculants, liquid, peat, and granular are the two or the three that are most common. Any of y'all have any experience with those? Anything good or bad? Yeah, yeah. And granular, yeah, he said granular is a lot easier to handle and that's by far the one that dominates most of the markets. And the difficulty with the, I mean, so you can throw those in a cedar box or they're pretty easy to apply with the seed. All it takes is just a little streak on the seeds and that's enough to get the rhizobia started. But I think sometimes people forget, you know, it's not like fertilizer, this is a live product. And so anything you do, if you leave it, you know, sitting out on the hood of the car or anything you can kill, and the liquid is just very sensitive to environmental conditions. So it's a lot harder to use. So granular is by far the most popular product. Nodulation typically takes place about six to eight weeks after, I want to see one slide here, okay. Six to eight weeks after planting, you start to see these nodules show up on the roots. And we never recommend any additional N when we, some people prefer that. And anyone have any thoughts why nitrogen would be problematic for this nitrogen fixation process? Exactly, yeah, plants get lazy. And so if the plant can take the nitrogen up from the soil, if there's a lot of nitrogen floating around in the soil, it's, energetically it's easier for it to just absorb it through the roots rather than going through the process of symbiosis. And so typically what we start to see is maybe 40 or 50 pounds of nitrogen in the soil and you start getting reduced nitrogen fixation. Because like I said, it's just energetically it's much easier. And so in fact adding nitrogen can be problematic if you're really trying to maximize your nitrogen fixation. So this one might be hard. No, it's not too bad. This is just a table of a bunch of different legumes. And it shows both the P fix in that first column there. That's the percent of the range in the percent of nitrogen in the plants that they've observed. And then how much that's in kilograms per hectare. So if you took 90% of that, that would be pounds per acre. But you can see there's a huge range. And so chickpeas, you know, between 8 and 82% of the nitrogen that is in the plants is fixed from the atmosphere. And then between 3 and 140 kilograms. So different species, different legumes have different capacity for fixing nitrogen. Peas are, let's see, the third, fourth one down there 23 to 73% and then 17 to 244 kilograms per hectare. So there's quite a lot of capacity for them to fix a lot of nitrogen. Fava beans are one that's thought to be pretty high, maybe the highest. And then in your warm seasons, generally a little bit lower. Ironically, soybeans don't, I mean that range that they show there, 0 to 450 kilograms is enormous. But soybeans aren't great nitrogen fixers relative to a lot of other pulses. And common beans are generally thought to be one of the worst nitrogen fixers. So this is just some studies out of Canada where they looked at the yield effect on different types of inoculants. And that one shows quite a dramatic increase from using the granular versus the peat and liquid. And in fact, the liquid, they really didn't find any difference than the uninoculated control. I'll say in our studies, we tend to see similar results from liquid and granular. But I think it's just a matter of how it's handled. And even in the shipping process, you know, to the wherever you buy it at co-op or whatever could have a big impact as well. So I'm not really promoting this type of assessment, but it gives me an opportunity to talk about a couple things when you're looking at nodules. So I said, if you want to check, if you want to go out and check your nodulation in your peas or any legume for that matter, there's a couple things to look for. You know, so four to six weeks, I would say six to eight weeks in a lot of cases for us is about the time you can go out and visually inspect. The nodulation process. And so these folks broke it down into three different categories. Obviously, there's the top. There's the above ground that you can see. I mean, it's pretty evident right away. If you're in a low nitrogen situation, your nodulation doesn't work for whatever reason. You'll start to see chlorotic plants pretty quickly. Chlorotic meaning yellowing, poor growth. But the nodule color and position. So that's one of the easiest ways to check these nodulation. And so in the process of fixing nitrogen, there's an enzyme, a small protein that the rhizobia used to break that triple bond. And that enzyme is very sensitive to oxygen. And so if there's too much oxygen in that surrounding environment, it deactivates the enzyme. And so what happens is the plant has developed a leg hemoglobin, which is the same as the hemoglobin in our blood. It does the same thing. Basically it attaches to those oxygens and it ferries it out of that environment. And so what I say that's the same is because it's got a similar iron content in the protein. And so it's red. It's that red color that's same as our blood. And so when you see nodules that are actively fixing, you can break them open and you see that bright red color. And so that's how you know it's healthy and it's working. Sometimes you see a greenish color, which usually means it's stressed. Maybe it's water or something else, maybe salt. And then there's white, which means it just hasn't started yet. And then there's black, which means it's dead. So those are kind of the four main colors that you see. Kind of gives you a sense for how well your plants are doing. And then nodule position. The ones that are thought to nodulate the most are those that are nearest the crown. So right at the, just below the soil level, you get these great big nodules that form around the crown. And then one thing we see with granular is you get a more diffuse nodulation. So on these lateral roots and some of these feeder roots, you see a little bit more spread in the nodulation, whereas the liquid seems to concentrate right in that crown area. And so you can kind of get a sense for where all these nodules are occurring within the plant. So we get this question every once in a while. So what happens if your nodulation fails? If your inoculant doesn't work, can you put nitrogen out there and rescue the plant? And there's a little bit of work out there. It's not all that convincing in my opinion. But this is showing on the left there, that left bar is zero in added and then uninoculated. So that's their control. And then there's nitrogen at seeding, nitrogen at four weeks, six weeks, eight weeks. And then the one on the farthest right is no in, but just inoculation. So a proper inoculation. And they found that you could maybe rescue, if you applied nitrogen at four to six weeks, you might be able to rescue it somewhat. But the problem with four to six weeks is what did I say about when you can start to see nodules in the plant? Four to six weeks. So, right, yeah. And they also, you know, that also in our country requires some sort of moisture to get the nitrogen down into the seed at that early time. And so it's not all that encouraging in my opinion. Okay. So some of the factors that affect nitrogen, I've kind of hinted at it already, but, you know, one, as I said, it's a very energy intensive process. Energy in the plant comes from ATP. And so the P in ATP is phosphorus. And so phosphorus is generally pretty important. We occasionally will put phosphorus down as a starter with the seed. Sometimes we let it go depending on cost. Obviously soil pH is a big problem. If you had to choose between high and low pH, lower pH is generally more detrimental to the rhizobia than higher pH. Where I am, we have a lot of calcareous soil, so we're seven to is pretty normal for us and much higher in some cases. And so as I said, there's a lot of iron in that in that nitrogenase that that enzyme. And so when you get into higher pH is iron is inhibited. And so that's one of the problems. Calcium is also important. So that's where you have problems with your low pH is and then high clay content. Tends to decrease nitrogen fixation mostly because of water. Either it gets waterlogged or we have a lot of inhibition by the clay for the roots to grow and expand. I think I'm going to skip this. This is I was talking about lazy, lazy nitrogen fixers, lazy, lazy bacteria as well. And all this is showing is that long-term fertilizer use tends to create lazier bacteria in some cases. Okay, so this is where I'm finally getting to some of our work. And what I'm going to be showing is from here in the central part of the state. And so the concept or the idea that we were tossing around with this is that if you buy an inoculant rhizobia from the store, it'll be the same species that they would sell anywhere. A company will sell one species, whether it's in North Carolina or North Dakota. It's the same one. And coming from a plant background, it didn't make sense. You'd never grow the same varieties of whatever corn that you would down there as you would up here or wheat or what have you. And so it doesn't make sense to us that they would just sell one variety. And so it turns out that rhizobia is everywhere in the soil. There's native rhizobia all over. And so we started to look around and we started to go to the prairies. I'm in the badlands and so there's pretty rough conditions. We wanted to go to places where there's a lot of stressful conditions. And we started culturing the rhizobia from all these different sources. And we just wanted to compare them and see, well, how do they do versus these commercial inoculants? And so one of the things that we were looking at a lot is drought stress. And so can we, we thought if we could go to places that experience a lot of drought stress. Well, maybe there's the bacteria that are in the soil there are adapted to that environment. And they might confer a bit more drought stress to the plants as well. And so what you're looking at there is a graph with there's 20, 21 and 2022. So there's two different years. And on the left-hand side of each of those is those numbers, the 23.2, 47 and 85 on the x-axis, those don't really mean anything. But those are just, those are native rhizobia that we tested them in the lab and they showed a lot of promise. And so then we compared them to a granular and a liquid and then an uninoculated control. And so there's basically two planning dates. So we didn't know if we were going to get drought when we tested these out in the field. So we wanted to just induce a stress on them. And so we had an on-time planning, which is the red boxes, and then the green one is a late planning. And so we thought, well, like I was talking earlier, if we push that flowering date into the growing season, we'll stress the plants. So we kind of an artificial stressor on them. And the trend that you can see is basically if you go from like this one 23.2, that's the total biomass at the time that we sampled the plants. You see it's roughly little over a thousand. 2021 was a pretty rough year for us, very dry, little over a thousand pounds of total biomass. And then in the second planting, it's about seven, maybe 800 or so. So roughly 30%, 40% reduction. Some of those, they drop a lot less, but they were also not as strong of producers. And so like that 23.2 is one that looks pretty promising. So if you look on the ones on the right, the granular, the liquid and the uninoculated, you can see those are kind of our controls. And remember what I said about those whiskers, that shows the spread of the data. So if you can see the granular, it goes from just above 500 to over 1250. And so there's tons of variability. And as a statistician, it's just a nightmare to try to make sense of a lot of these data. But the idea though, and I'll admit we had some varying success with this, the idea though is that if these are more drought tolerant, we should see a smaller drop off between our plant one and our plant two dates. So let's, this is just biomass. And I'm going to talk more about the nitrogen fixation process. So the graph, these two graphs are showing the amount of nitrogen in the plant that was derived from the atmosphere. And so the scales are totally different between these two. In 2021, as I said, pretty rough year, we fixed somewhere between zero and 50% of the nitrogen. So again, of all the nitrogen that's in the plant, that's how much came from the atmosphere, meaning the rest, the plant just took up from the soil. And so look at those lines within the boxes, that's the averages. And so in general, we have some pretty good looking results, especially in 2021 where we saw, I wish I had a pointer, typically where the green bars, where those averages are, that's what we're looking at versus the ones on the right hand side where the controls are. So we still see that they're fixing nitrogen under very stressful conditions, which is good. It's encouraging. In 2022, we didn't see quite the increase that we saw in years prior in better conditions. So you can see though, in a better growing conditions, we went from maybe 20 to 50% fixation to now we're up 70, 80, even a little bit above 80. So now the plants are getting 80% of the nitrogen from the atmosphere. So inadvertently in this process, we found some rhizobia that can do pretty well, even under more ideal conditions when actually we were just looking for ones that can survive. Okay. And so this is total nitrogen fixation. Again, pay attention to the scales, they're vastly different. So in 2021 under drought conditions, even our best performing ones were only fixing 10, 15, 20 pounds of nitrogen. So the range is not very high to begin with, but we saw a couple pounds difference, you know, so I don't know, for whatever that's worth. But then if you look at 2022, now they're fixing over 100 pounds of nitrogen. And this is where we fix, you know, our better performing ones, we're fixing 30 or 40% more nitrogen in the good conditions. Now when we drop and we stress them out, they didn't change much. And so it was not what we were hoping for, but it's a good result in the sense that we know that we can make better rhizobia. So if we have native soils, then what's out there? Or if nothing else, we can make something that's on par with what they consider their elite strains. And then we started to do a lot more work with salinity, high salinity areas. And so we're breeding, it's weird to say, we're breeding rhizobia to perform well in higher salt content. And so we did the same process. We just went to saline seeps and we take soils and we start to culture the bacteria out of that and see what's in there, really. And so this is just a graph showing how sensitive peas are to salt. So you can see it's a pretty linear, linear decrease on the x-axis. There's increasing content on the y-axis is the percent of normal. And so it just drops pretty rapidly whereas wheat, and in particular barley is one that is pretty salt tolerant. But wheat, you can see it kind of hangs on for a little while with increasing salt and then it drops off. So there's a little bit more cushion in wheat than there is in peas. And this is just, I don't have any plant or field data for this yet. It's still in the lab, but this is just showing the range. So each one of those blue, orange and gray bars is one variety with just higher salt contents. And what it's showing is like the one on the left there. In good conditions, it's all right. That just shows the growth. And so the second you start to add any salt, it just dies immediately. Whereas the one next to it, you can see it hangs on even through higher salt conditions. It's still able to grow. And so what this is highlighting really is just if you look at a bunch of a panel of different rhizobia, there's a huge range in terms of salt sensitivity. Does that make sense? Okay. And this is just at even higher levels where it's like a seawater. And so some of them do, so we screened probably, I don't know, a hundred different species of rhizobia. And so these are some of the better performing ones. And this is kind of what we did with the previous test is how we screened the rhizobia in the lab to say, okay, let's take this group to the field. Yeah. So is this commercially available? Can I call it and say I would like to buy salt powered rhizobia? No, no. The funny thing though is this is out just in a field somewhere. These are all just native rhizobia that we found in various fields. And so to my knowledge, there's nobody that has a range and says, okay, yeah, so what are your growing conditions? Okay, this is the one that I would recommend, like they would with wheat. You know, they don't have that, that available. And that's what frustrated us and kind of what we were trying to work for. That's a great segue. And so what we're really big on is this DIY microbiology because the whole thing was we thought like, well, what if we started making inoculants, whether it's rhizobia or whether it's some other inoculant from farmers' fields for farmers' fields? You know, why do we need to have this exogenous product? Well, maybe we could make it right here at home. And so this whole thing was kind of a demonstration or a pilot to say, yeah, we can find these bacteria right in our backyard that might be just as well as anything you can find commercially. And so we're really big on figuring out, like, okay, so what can we actually do? What could a farmer do in their shop? How far could they actually get to making their own inoculant if they were so inclined? And honestly, we don't get a lot of farmers saying, I want to make my own inoculants. But maybe it's because we, you know, they didn't know it was available. I don't know. So anyway, the things that we do in our lab, that's kind of the guiding principles. How much can we do just on a very low tech? And so what I'm showing there is a lot of the standard things that you'll see in a microbiology lab. For example, you'll always have autoclaves, big, great, really expensive autoclaves. And so that's to sterilize everything. You know, obviously you need a very sterile environment. And so we were trying out instant pots. I don't know if you use those at home. But it turns out they're a great autoclave. They get pretty high pressure. And it's not quite up to the standard of an autoclave in a lab. But it's close enough where you don't have, it sterilizes everything. And so it does pretty good. We use it for almost everything. In the bottom is a clean hood. It's kind of hard to see in that picture, but we just built. And we put this up online just, you know, like how to build a clean hood. Which is, you know, basically an area that you can put your hands in and you can work in a sterile environment. It's got a fan that blows out and it's got, it's got, I guess it'd be negative pressure on the inside. So it keeps air out and doesn't let anything float in. And then we've been trying a lot of different growth media. And so on the standard, on this left-hand side. So that standard growth media is basically our recipe. When we want to grow the microbes in the lab, that's our recipe. You know, salt, magnesium sulfate, yeast, potassium phosphate, mannitol and DDI water. So just really pure water. And so we sent the, all the, all of our interns went to the grocery store and made up their own recipe. And so we tested them out, trying to grow the stuff in a media that you could buy from the grocery store. And so this is the one that, one in our lab was table salt, baker's yeast, epsom salt, sugar and pediolite. And so we had a lot with Gatorade and all kinds of different stuff. Creamer is so full of, like the process creamer, it's got an unimaginable amount of stuff in it. But you can, you can grow, you can grow a lot of bacteria in it too. It's got, it's not so bad that you can, it'll kill all the bacteria. And then we're even looking at, so nanopore is a newer sequencing technology, genomic sequencing technology. And so their whole goal is to bring, is to bring genomic sequencing to the masses. And so we're getting to the point where, for example, in the summer I was in the Amazon rainforest and we were sequencing all kinds of stuff in a very makeshift lab out in the middle of nowhere. We were sequencing bats and snakes and plants. So it's not quite there where, you know, any farmer is going to have a genomic sequencer in their, in their shop. But it's getting to the point where we could make this a more distributed thing where maybe somebody sets up a lab and we have people send in their soils and they can start making more, you know, making tailor made inoculants and things from farmer's fields. And that's kind of our, that's kind of one of our end goals. And again, I don't know if farmers are actually even interested in that or not, but I think it's a, I think it's an exciting process. And I think it's, it's just nice to show that the step doesn't have to be, you know, secured away in a lab. We can do a lot on our own. And then I'm just going to end. And so the last thing this is kind of a side note is we've been doing a lot more research on winter peas. They're starting to make their way out of the Pacific Northwest and breeding some new varieties that are more winter tolerant. I'm a big fan of fall planting if possible. And so these are planted like winter wheat is. And this is two years ago. Yields were not great only in the 20s, but that was our spring pea average. And so it's right in the ballpark. Some of them are right there. So we're in the kind of the early phases of doing this and screening varieties that will survive in our winters. You know, it's debatable whether North Dakota and South Dakota winners are the same. We're kind of in the banana belt over in the West. And so it's always a little bit warmer. There's certainly a lot of possibilities. And I think having that extra growing season for the peas and just from, you know, like the region egg talk. That's three extra months of a root growing in the ground. And so I'm a huge fan of these. And this is just one area that we're doing a lot of work on protein is pretty much the same, even a little bit higher. So I'll go ahead and break there. You got five minutes, but we're the last session. Okay. Yeah. Well, I've talked to probably enough, but you know, this is just some acknowledgments. This is a Sarah project that we're doing a lot of the riso be a work on. Most of you maybe are familiar with them. It's a USDA branch. And then the pulse growers from South Dakota have a council that funds some of our work. And so there's all my contact is just my first and last name at SD state. But if there's any questions now, be happy to chat. Yeah. And it's a double-edged sword really. So yeah, you do get some residence time. You're talking about from an inoculant or just your. Yeah. Right. Yeah. And so there's been a lot of study on that. And there's two, two things that they're studying. One is if I inoculated two years ago, well, some of that risobius still be in the ground. And the answer is maybe depending on the conditions, you know, it's a typically say, well, inoculant is cheap enough that you might as well inoculate anyway. But two, does it attract the native populations? And that I don't have a good answer for. I don't know that you increase or decrease. And we never really studied the effect of the plant in drawing out the rhizobia. So that's a good question. And I think, you know, certainly the root exudates that are in, you know, that stay out there will, you think would grow the population. Now at the same time, these plants we talked about are finicky and they're susceptible to a lot of diseases. And so the thing that you often grow with in terms of bacteria is fusarium or some of the fungi. And so planting them too often, you know, you start to build up a lot of Montana growers. And even in North Dakota, we've seen it where, you know, they just can't grow peas anymore because disease buildup is so high. And so I don't know that I would even recommend it at least. We always say four to six years in a rotation. And that's a big debate. And I think, you know, so like a lot of people went from, they went from the wheat fallow system to a wheat pea system. And that led to a train wreck in a lot of cases. So what's the magic number? I don't know. I mean, of course, it's going to be context specific, how much precipitation you're getting and a lot of different things. In fact, you know, the farther west where it's drier, I think it's a bit less conducive for some of those things, like some of the bacteria and some of the fungal diseases. But you've got to kind of thread the needle. And if you've grown them, you know that pretty well, I'm sure. Some of them. Yeah. Yeah. And that's why, you know, chickpeas, we think will never work much beyond the river. It just gets too humid for a lot of those. Unless the breeding really ramps up and they get better, better varieties. Any other questions? No, please. Yeah. It's a super simple thing. We just use the plant to trap them. And so we'll take a soil and we just grow peas in it. And then we pull out the nodule. They'll nodulate naturally. And so then we just excise the nodules and we kind of crush it up and make a soup of the nodule. And then we can grow the nodule on those media that I talked about. So really the plant does all the work for us. Because so many of the bacteria in the soil we can't culture, we just can't figure out the right conditions. But in this case, you know, it's actually a pretty simple process. It's interesting though that when we, sometimes we'll sequence the nodules and there's rhizobia, but there might be 20 other things in there. And it's a big debate about what are all these other bacteria doing in the nodule? You know, are they freeloaders? Are they just eating the carbon that's coming down? Are they helping somehow? We don't really know, but when we sequence those things, there's always a bunch of other stuff in there. So that's one of the tricky parts is how do you make sure, let's say you were to grow your own, make your own. How do you know that you have a pure culture of the rhizobia? And that's a bit tricky to figure out. But there are certain ways that we can kind of at least hedge our bets anyway.