Seed Inoculation and Microbial Biology: What Microbes Do for Stand Establishment

0:00 All right I think we'll go ahead and get started here. I want to welcome everybody to our fourth webinar of our biological webinar series. Just some housekeeping notes that everybody is muted. If you have questions you can type them into the question box and at the end of our presenters presentation we'll go through those as many of those questions as we can. And so with that, this will also be recorded and put on our YouTube page for later viewing as well. So with that Keith I'll let you know today.

0:40 All right thanks Dylan I appreciate that. Well hello folks and welcome again to our webinar series. We appreciate everyone joining us. Our guest today is Mr David Olson from California. Now David is relatively new to our network of experts that we've been working with and he'll explain a little bit of his background, but he's certainly not new to the whole field of microbes and microbial amendments for the soil. David is a sixth generation farmer from California so they've been farming out there since 1850, so he's got a long rich heritage of working the land and the soil and preserving that. Obviously they've done a great job of preserving that for their generations. He was mentioning earlier that his dad actually was one of the first ones to work within what would later become IPM, the integrated Pest Management side of things. So he comes from a long line of innovative farmers who have taken what they've learned and have turned that into things that other people can benefit from. His company Sustainable Growing Solutions produces different inputs, microbial and biological inputs, and they are one of the suppliers of some of the key ingredients that we're using through our Elevate AG products and that is ending up on our seed as some of our seed treatments as well. So many of these things are coming from David's company, from David's experience and all of the things that he's learned over the years. So with that I'm going to turn it over to David. He's going to be giving us a lot of information about soil biology and how the microbes affect all that. So David, Jonathan or Dylan and I are going to kind of hide ourselves and go ahead and share your screen and take over.

2:40 Well thank you very much for the introduction Keith, appreciate that very much. Okay and did that come across? Yep yep there you go, we can see you good now. Good.

2:52 Well thank you. The microbiology and soils and plants is a huge topic so what I really wanted to do was focus today on the importance of the roles that microbes play in seed germination and stand establishment. So that'll be kind of the focus of what we'll talk about. So we'll talk about what are the differences between a healthy soil and just kind of an average production soil and why is that a problem. Talk about again the functions of the microbes and seed germination and why that's important. Talk about some of the different inoculation options and methods that we have for you and show you some example crop programs.

3:34 It's a little bit of background. So plants and microbes co-evolved. They have worked together in an intricate system. So their interactions are bidirectional and profound and intricate. So when we're missing our soil biology we're really missing out on all the benefits of what the microbes do for the plant. Something that I think dictates some of our thought process is that when you have a plant or a seed you have a certain inherent genetic potential yield that the plant has. And really as a farmer our job is to mess that up as little as possible. And when I hear a grower say that they're going to make yield it kind of makes me cringe a little bit because that implies that you can force the plant to do something that it otherwise would not have done. And that's almost backwards from the appropriate or I think the more productive relationship, which is that we're trying to be the stewards of the plant to make sure that it has everything that it needs in the right quantities at the right time. And if we do that, working with the biology to deliver that, things growing actually gets a lot simpler and we have a lot less problems and we tend to have a lot higher yields.

4:56 So when we're working with the microbes, we build the micro population, make sure they've got the building blocks to deliver to the plant and then the plant puts out exudates which direct the biology to deliver what it needs when it needs it. So that's kind of the overarching philosophy. So a lot of the problems that we see are things like over fertilization. So again the mindset of it going to make that plant have yield if you put on too much of anything it really gets in the way.

5:27 Way of things and it ends up building weaker tissues that are less robust and more vulnerable to pests and diseases. So as we can back off on our fertilizer inputs and rely more on the biology to deliver what the plant's asking for, we have a lot of those problems go away.

5:47 The process is pretty easy. You build a large and diverse high functioning micro population, you make sure they have all the nutrients they need to deliver the plant, cut back the fertility a bit, and you can do that by monitoring with sap analysis. Sap analysis gives you kind of a pretty close to real-time assessment of the nutrient status of the plant, so it gives us a chance to course correct and so it's not like we're flying without a net as we start to cut back on some of these fertilizer inputs.

6:20 Microbes will compensate for a lot. If there's a base saturation problem with the chemistry of the soil, they can compensate for some of that, deliver more balanced nutrient status in the plant. Same goes for plant hormones that they will deliver plant hormones to the plant, but not more than what the plant wants.

6:40 Here's just a quick example or illustration of how microbes delivered nutrients to the plant. What you're looking at here on the left is the tip of a root hair and so the blue are the cells of the root and the green are microbes, and you can see that there's more microbes than there are plant cells. As I mentioned, the plant puts out root exodates in the form of sugars, and sugars of course are carbon, and that's the core of the carbon cycle in the soil, and carbon sequestration happens through that mechanism.

7:17 As the plant is feeding the microbes, it also puts signalers into that which direct the microbes to deliver what it wants. So we knew that this was happening for quite some time, but only recently has science been able to explain exactly the mechanism of how this is happening. So in the very end of the root hair, there's basically a portal, and the microbes go into that tip. Inside the tip of that root is a superoxidase compound which literally melts the shell off of the outside of the microbe. The microbe goes into the cell of the root, and so now it's in the cytoplasm of the root and it's delivering its nutrients directly to that cell.

8:02 Once that exchange has happened and there probably is more going on there than we even currently understand, but at least we do know that this is happening. After they do that process, then they're ejected back out. They reform their cells, they collect more nutrients, and they take the right again. This is why it's important to have a great soil biology, because without it you're missing out on this.

8:30 Here are all the functions that the microbes do for the plant. They fix nitrogen, they cycle nitrogen, and they deliver nitrogen to the plant in the form of amino acid, which is much more energy and water efficient for the plant and less susceptible to encouraging certain types of pests and diseases. They also solubilize phosphorus and potassium, making sure that the plant actively gets delivered those more efficiently.

8:59 There are some fungi that are endophytes, meaning that they have part of their body inside the root, but they will have a hyphae that will go out and act like a root hair except for it's about 200 times smaller than a root hair. Some of those actively collect things like phosphorus and potassium, and what they're doing is they're mining interstitial spaces in the soil that the root hairs can't get to. By having this relationship of these microbes, especially the fungi with the roots, you can double or triple the effective volume of soil that those roots are mining.

9:42 Of course the microbes are translating the nutrient potential of the soil into actual nutrient availability for the plants. So they're doing things like chelating cations so all of those cations can be pulled out of unavailable form and put into plant available form by the microbes. Whereas microbes are also part of the immune system of the plants, so they suppress diseases either through direct predation, quorum suppression, which is a huge topic that I can talk about an hour on just by itself. They also compete for food and habitat, so they're a great part of the protection system of the plant. Without them, they're quite vulnerable to all sorts of diseases.

10:24 They also increase the sap and sugars in the plant and make the plant less attractive to biting and sucking insects. So there's all sorts of reductions in our pesticide bill as we get the soil biology built up, and of course the biology also delivers all sorts of plant.

10:47 Growth regulators and so if you think about all the different physiological stages that a plant goes through through its life cycle, you know every one of those life cycle stages where switches from vegetative to reproductive, there's a whole bunch of hormonal things happening. Plant biology in the soil contributing those hormones to the plant makes the plant much much more efficient in making all those changes. So those changes make a difference in what nutrients the plant wants, what the ratios of those nutrients are, where they're being delivered to within the plant and what tissues are being built at the time. So really profound differences in what's happening with plant and all dictated by these different types of plant growth regulators. All this goes much much better as these are being supplied by the biology to the plants.

11:37 So let's talk about what a good optimal soil would look like. That should have about a billion microbes in a gram, so something that takes up that much space has a billion microbes in it. And should have up to about a thousand different microbe species in it. And of course in the world we don't even know how many species of microbes there are, but it's popularly believed easily in the multiple millions of different species. So there's lots of different microbes. The problem is with production soils, many soils have a thousandth to a millionth of what a healthy population would be. They're just beat up and you can imagine in any kind of natural system where things have a huge set of interdependencies, if you have a thousandth of what a normal population is, it's broken, it's not working anymore.

12:30 So our overall arching objective in our crop programs and in our products are to restore all of the soil biology. So we have a very broad spectrum set of biology that we bring in our products, which is very different from how most other biological products function. Even in a field that's been farmed regeneratively, a lot of the time what we find is a good established bacterial population, but oftentimes very lacking still in the fungal population. And that's another thing that we do with our programs is to bring up that other set of biology back, that has so many things to contribute.

13:14 So what I did is put together kind of a series of Venn diagrams here just to kind of conceptually represent what a healthy soil would be like versus what production soils look like in several scenarios. So if you visualize this, you know large dark circle is the darkest color means that there's a high population size. And the larger size of the circle just means that there's more species and a higher sense of diversity. So use this as kind of your frame of reference of what optimal would be.

13:52 So in a typical production soil, instead of a billion microbes per gram, we might have a million. So that's one thousandth of that optimal soil. And instead of a thousand species, we might only have a hundred, so it's a tenth of what the species diversity would be. And then this is actually maybe even slightly optimistic in a lot of cases and pretty much assumes that we have a decent set of crop rotations and even to have this kind of soil.

14:22 Here's kind of a visualization of why we do crop rotations. So the larger bubble there you see, that's slightly darker, that's what the crop that's growing there now and the microbes that are associated with that plant in the soil. To the left of that is a smaller lighter circle of the crop, kind of the residual micro population that's there from the crop from two years ago. And then so on with last year's crop. And of course as we're doing a crop rotation, you want to build more biology in the soil, but those parts of the circle that are not overlapping, you are hoping that is basically where your pathogens would be. Because if the pathogens are in the area that the circles intersect, then you're actually carrying that pathogen around the rotation. Of course, it's one of the reasons why we do rotations is to not have that happen.

15:31 Now we look at a lot of soils that have either effective monocultures or GMO based genetics for the plants and we find that we have even a much more severely affected biology. So now the circle is much smaller because we have less species and it's less dark colored because there's much smaller population. So we have a millionth the size of an optimal soil.

16:02 Terms of population and a twentieth of the size of population diversity. So at this point you're treating your soil like it's basically a sponge to put stuff in for the plant to pick up and it's just physically to hold up the plant from falling over because it's effectively a dead soil that's not really doing any beneficial functions for your plants.

16:26 Yeah, the idea of having a cover crop is you know here we have different species or hopefully even different genus of cover crops and different microbes that associate with each one of those different species. So now by having a diversity of plants to colonize and have complementary soil biology with, now we have 10 million microbes per gram of soil which is only a hundredth of what the optimal soil is but it's certainly way better than the other options and now we have a hundred species of different microbes. So you can see again the more different, or the more diversity there is say in the genus of your cover crops, probably the less overlap you're going to have in these circles and so the more effective building of the soil biology and function you're going to get out of that.

17:23 Now here's a conceptual representation of those same cover crops but now inoculated. So we have some seed inoculant treatments that we have that have hundreds of different species of microbes. So you see that each one of those circles got bigger because they have more species that now have colonized them. They're also darker because we now have higher populations of those and then we have kind of this over background circle which we've built more biology that's just inherently in the soil and not necessarily associated with the plants themselves. But now we've got a tenth of what an optimal population would be rather than a hundredths and so we've got a much better situation.

18:11 So this is what we're after here. So as you get your cash crop and you interlace this with the biology that's been built by the cover crop and the inoculated seeds, this is what we're trying to accomplish.

18:29 Here are all the different opportunities to kind of work biology into your production program but not all of these are for everyone. But you kind of look and pick on the ones that work the best for your operations. So we can inoculate the soil at bed prep. So if you're listing up and doing any pre-plant fertilizers, that's an opportunity to get biology going in the soil. The more time you give biology to do its job for the plants, the more effective it is.

19:00 We're going to spend a lot of our time talking about the seed inoculation part of it here so I won't go into it more. But again, the more time the biology has to work for the crop the better. So seed inoculation being that first opportunity, so it's a critical one and makes a big difference. Then we have transplant drenches and over the seed line. Between seed inoculation and planting over the seed line is probably the best value in terms of bang for the buck in getting a lot of value out of the micro materials you're putting on and the response that you'll get out of the crop from it.

19:39 And we have various other opportunities like side dressing or top dressing, irrigation applied, foliar, and also crop decomposition which is also another excellent one for bang for your buck. Does a ton of stuff for you but we probably won't go in any detail on that today.

19:59 So microbes do a ton of stuff for the seed germination and stand establishment. And that interestingly, the first relationship between the microbes and the seeds is the fact that the seeds actually have a lot of microbes in them. Some recent university research has indicated that as many as 2,000 different microbe species have been discovered and documented inside seeds. So if you want to have a strong plant it really needs to come from strong seeds. Strong seeds come from plants that are vigorous and have a healthy microbiome associated with them. So when you see seeds that have microbiology in them, that actually occurs during the pollination process. So you want to have a good healthy biology in the canopy of the parent plant as they're setting those seeds so that you have seeds that have good biology to start with.

21:00 So when we get to the germination process, here's what's going on. So when the seeds first starts to hydrate, one of the things it does is it puts out an exudate through the skin of the seed and it signals the microbes to come colonize it. And so it's literally an invitation, it's putting out food for them to kind of participate. So when they do, the microbes come and they provide a lot of different metabolites and enzymes. So the first thing they do is they provide some enzymes that soften the seed coat.

21:31 Allows moisture to get into the seed more efficiently. They provide amylase enzyme which converts the endosperm of the seed into the starch and the endosperm into sugars, and it uses that energy for the seed to germinate and grow.

21:48 Then there's a bunch of hormone things that are happening. Some of the microbes do produce these hormones and those hormones are used by the seeded in the germination process.

21:59 Now this is very complex and I'm very greatly simplifying and generalizing about this but jasmonic acid actually helps the seed come out of dormancy. It's the signaler to start the germination process. It does that in collaboration or in conjunction with also gibberellic acid. And oh, the jasmonic acid also is responsible for the primary root initiation, so super super important because that's basically initiating the tap root.

22:32 Indole acetic acid cell division, so again that's the spark of the seed to build itself into a plant. Gibberellic acid is responsible for cell elongation. So if you think about that cotyledon and its strength of pushing through the crust in the soil, gibberellic acid gives it a lot more strength to push through a crust. So you see a much stronger, faster emergence because of that.

23:01 The microbes of course are also feeding the seed already the amino acids that I talked about, the phosphorus and the cations. They're also feeding other root related and growth related hormones like auxins and cytokinins. And as I mentioned, the endophytic fungi, the ones that partially live inside the root or colonizing the roots at that point, and they're extending the root zone more quickly.

23:31 So if you think about your fertilizer placement relative to your seed location, the endophyte fungi are going to help that plant get access to that nutrient set much more quickly.

23:44 So what happens when we have that level of micro participation in seed germination? Well, we have an increased percent of seeds that actually germinate. There are quite a number of studies that have been done on that. We get a faster and more uniform germination, so we have less lag. I'll talk about why that's important in a minute. We get a better stand establishment, more uniform. We get a stronger push through the crust, so we have less emergence failure. We have less mortality due to damping off and other types of diseases. You get larger root systems faster, which mine nutrients more effectively and efficiently as well as water, so it makes our seedlings say less drought.

24:32 One thing that happens here to us and some of our field crops here is we'll have the appropriate moisture in the soil for planting and then we'll get a very very dry north wind and all of a sudden it'll pull the moisture out of the first couple or three inches of soil. So in some cases that can really really set back a crop, but if we've got a stand that has more deeply rooted itself faster, it's much less vulnerable to that. I'm sure many of you have similar types of challenges.

25:03 So of course with all the above, you get a higher stand population, you get better nutrient status, and you get a more vigorous plant.

25:11 So when we're talking about lag and emergence, and this there's quite a bit of variability from crop to crop, and so again this is pretty highly generalized, but we did some current emergent studies a couple years ago where we're looking at the time lag between the first seed that emerges and how many of those emerge within the first 24 hours of that happening, and then how many emergences happen the second 24 hours and the third 24 hours. And what we saw was 17 to more seeds or more plants germinated in that first 24-hour set, and less than the second 24 hour, and much less in the third 24-hour set. So we're kind of moving everything forward to much more uniform.

25:55 Well, and again it varies from crop to crop, but there's a lot of different research on this, but a five to ten percent incremental loss and yield for every 24 hours of delay for emergence. So right there, remember what I said is a plant has a certain amount of genetic potential for yield. Our job is to screw that up as little as possible. So if we have made the emergence much more uniform, we've already screwed up less. And things happen really fast with a crop. A lot of the principal things that define what your yields are going to be happen very very early in the season. So if we have inefficiencies or imperfections in what the status or condition of the plant is, we really get dinged for it hard out of the gate.

26:45 Inoculation, so we have several different options for doing seed inoculations. Again, pick the one that works the best for you. We have a dry inoculant powder called foundational fungi and it's really a neat product because it is truly fungal dominant and it comes from a different set of biology than pretty much any other product that we know of. So it's very complementary to some of our other products and it does bring that fungal component which is missing from even some of the best regenerative soils that we've seen.

27:20 Scene inoculation is probably one of your best bangs for your buck that you can get because you use so little material and because it gets to participate in that germination process it has so much more opportunity to beneficially affect the outcome of a crop. So we're only using two to four ounces of this product per 100 weighted seed. There's any number of opportunities you can put it on—you can put it on during handling or bagging or even just pour it over the hopper on the planter.

27:51 So typically you might be looking at anywhere from two to eight dollars per acre for something like that or for either one of these options for the inoculation. So if the powder's not right for you, we also have liquid inoculants, so these are again very high populations, very diverse biology. The metagro F is actually the liquid version of the powdered foundational fungi, so same biology just in a liquid form. And then we have the metagro 5X plus, which is our concentrate biology. It's a different set of biology than either the metagro F or some of our other products, but of course then we've got some options for how we apply liquid products there.

28:32 So we've done a number of studies about what the biology does. We're actually trying to re-establish the biology of the soil. So we go from a very impaired production soil and then we do some inoculants on it. So in this case, this is a study where we looked at the base condition of the soil and its beneficial functions that the microbes are supposed to do for the plant. And then we did one treatment of our biology and then we came back and said, okay, how much have we actually improved the function of the soil? Because we wanted to quantify that.

29:04 So we had 190 percent increase in nitrogen fixing, 420 percent increase in phosphate solubilizing—so really making some big differences there. Double the species diversity. 73 percent colonization rate, so of all the different biology that we have identified in our product, 73 percent of that we actually were able to find in soil afterwards. The cation chelation, I've talked about that, so 270 percent increase in that function in the soil. 190 percent increase in the plant growth regulators. 500 time increase in the systemic acquired resistance or the induced systemic resistance functions of the biology. So in this soil that function was nearly absent, and here we brought it back to, I think, a very highly functioning level.

29:58 So what it does is of course makes your plant much more resilient to pests, diseases, toxins, stress—you know, heat or cold stress or drought. So then a 540 time increase in chemical residue degraders, so breaking down all those old chemical materials in the soil. 140 time increase in organic matter decomposers, so naturally we're having our crop residue break down much more quickly. A 93 percent reduction in plant pathogens, so in this case fusarium was the dominant fungal pathogen and in the subsequent testing it was almost a non-detect. And then a 99 percent reduction in anaerobic microbes.

30:45 So my apologies for making this slide sideways, everybody—tilt your head to the left a little. And so you've heard me just discuss at length the fact that we can actually restore the function of the biology in the soil, but the critical thing is what does that actually translate to in terms of plants? How do they respond to it? Well, this is a sap analysis from a couple years ago with labs. And what we did is we had the treated—the grower standard program—and then we had our biological program. And in the biological program we cut the nitrogen back by 50 percent, so way less nitrogen.

31:33 So we're testing a lot of things here. It's like, okay, so everything else is the same except for nitrogen. What was the nutrient status of the plant? How much additional nutrient were we able to get into that plant because of the function of the biology we've established here? So at the bottom of the page here is three bars or three sets of bars that look at nitrogen. So the middle one is total nitrogen, and so that's going to represent amino acid form of nitrogen predominantly in this case. And so even though we put on 50 percent less nitrogen.

32:09 Nitrogen in the program, we had 20 percent higher total nitrogen status in the plant, which is excellent. That means that not only we save all that money on the fertilizer, which was more than enough to pay for the biological program, but we have a healthier plant in a higher protein content plant. And the other part that's good about the nitrogen set here is we had a substantial reduction in the nitrate concentration in the form of nitrogen in the plant, so a 56 percent reduction there. And that means that with less nitrate concentration, there's a bunch of biting and sucking type insects that will be much, much, much less interested in feeding on this plant.

32:54 And then the ammonium form has almost disappeared of nitrogen, and ammonium forms in a plant when a plant is under stress. So you know how we say that mites like stress plants? Well, the reason why it might select stress plants is because they have higher ammonium in them. By nearly eliminating the ammonium form of nitrogen in this plant, that means that mites are not going to be interested in this plant at all.

33:24 Then looking at the very top set of bars is our total sugars. So we've increased that by 272 percent. So the sugars are the carbohydrates that are formed by the plant from photosynthesis. So we've increased the photosynthetic efficiency of the plant, but it also means that we're building and storing more energy in that plant in the form of those sugars. Those sugars being in the sap also as a deterrent for insects and insect pests because insects don't have a pancreas, they can't process sugar. So as you get the sugars up in the sap again, much, much less interest from pests and bothering that plant.

34:03 Then pretty much across the board, you can see for all of our principal nutrients here we have moved the needle a lot. A 29 percent increase in phosphorus. You know, I have a lot of growers that complain to us, and it kind of draws a lot of people into a biological program with us, that every year they take tissue samples on their crops and they see that the phosphorus is not at the target levels that they want, and every year they look at their soils and they have tons of phosphorus in the soil. And the PCA comes back every year and tells them to put on more phosphorus, and they know that that's not the solution because that's what they did last year. So by having the biology there, we're actually getting the biology to get the nutrient potential of the soil successfully into the plant. And so we're seeing that across the board here in all these sap analysis.

34:58 But again, if we saw something that was lagging in nutrient status, especially as it comes up on an important plant growth stage which relies on that nutrient, this tool gives us the opportunity to intervene and put on a foliar and address that lagging nutrient content.

35:19 So what makes our biological products a little different than everybody else? Most other biological products just have a handful of microbes in it. So if you remember those Venn diagrams that I was pointing to earlier, if you have say a product that has a half dozen species in it, it's not going to change those diagrams at all. So an acid inoculant that has a half dozen species might give you some of those functions, but it's not going to rebuild your soil or rebuild the function of your soil.

35:49 So our approach is to have a very large and very diverse set of biology. So we have up to 20,000 different species and 100 billion microbes per milliliter. So that means for every drop of our product, you have 5 billion microbes in it. So very, very different. And our objective is to restore all of the soil biology, not just a handful of species. And for every one of those plant beneficial functions that I listed before, I don't want to just have one microbe, I want does it do it.

36:24 Our products are shelf stable, which means that you can store them for years, and you can also mix them with synthetic fertilizers with no or very little mortality. And there are other approaches that I think is different is that we take, we look at the entire crop cycle and all the different growth stages, and instead of just focusing on our own products, we integrate our products with some other products to build a comprehensive crop program. Because again, we're trying to get the biology there and functioning and then delivering what the plant needs at the time.

37:03 So here's just an example of what a crop program looks like, and again, not every opportunity fits in every operation, so these are all optional. Obviously they work better together. The more of them you can do, the more benefit you get. Some give you a much better response than others. Some of them are better if they're in combination. So we already talked about the seed treatment. In this case I'm talking about the powder foundational fungi. We've already talked about what the benefits are of that.

43:39 Around right now on the plant so it's almost like the difference between you know the sap analysis it was kind of like a blood test. And the analogy that I make between the two is this: the tissue analysis is like trying to drive your car by only looking at the rear view mirror versus a sap analysis at least you can see the front bumper. So instead of being reactive we can be a little bit more proactive with it especially if you're taking successive samples and looking at the trend lines of what the nutrients are and then being cognizant of what the peak nutrient demands are for each crop growth stage you can really pay attention to the right nutrient at the right time and then make sure that the plant's getting at the right quantity so that's what allows you to use this as more of a proactive tool rather than being reactive and kind of being behind the curve.

44:37 Yeah and they got a question here from Stephen. Should we be making an extract and spraying it on the seeds or is the inoculant that you know we at Green cover or are putting on the seed is that sufficient? So maybe is can you get too much on there? Is there going to be any different effects?

44:58 Yeah as long as you know what the characteristics of the material you're putting on I'm always in favor of greater populations and greater diversity. So if you have say your own compost tea or something that you're doing I think that definitely would complement everything that I was talking about.

45:23 So our and then and your products where do those microbes come from? Do they come from California and maybe would microbes from your own farm ground be better suited for your you know whoever your contacts is wherever you're located? Yeah that's a great question because there's a lot of perception around that. A couple aspects to it: one is that we our microbes come from several a bunch of different sources. We use earthworm cultures and some various mother cultures that we've constructed by collecting biological and soil samples from literally all over the world. So we've collected biology in 17 different distinct different kind of biological conditions and so then we continue to nurture and propagate those over time and so we've really built up some unique biologies. So in addition to the earthworms we also have another set of biology that comes from a different trophic level of animal on the soil which brings it an entirely different set of biology and it's very complementary. So when our diversity in our product is I think second to none out there.

46:50 But then the other way that I would respond to the question is all biology is everywhere all the time. The only thing that varies is the population size and it's representation of the population because with a single particle of dust you can have thousands of microbes on it and if you have a dust devil or something like that that picks up a bunch of dust the dust gets up in the air in the jet stream loaded that particle of dust can be anywhere in the world in three days. And then as it falls out of the jet stream you can think of it as we have this constant rain of these dust particles that are from all over the world that are raining down around us. So this the biology of the soil you know on your farm may be unique to your conditions based off of the combinations of history of what's happened to it but the biology is not unique to anywhere else.

47:49 Okay, very good. I guess I would say one thing about biology and fitness that lab raised microbes are much less fit and successful at colonizing in the soil and performing their identified beneficial function than a community-raised microbe. So if somebody's got a compost tea or something like that you may have the same microbes in that or some of them maybe the same as what's on some microbinocular label but the ones that are grown as a large culture I think are going to be much more suited much more likely to successfully colonize been much more likely to do their beneficial function than something that's been raised in a lab in a Petri dish in isolation from like all other species.

48:43 Okay, very good. What makes your products shelf stable and is that just you know in the jug or how does that relate to being on the shelf or even being on the seed? You know what makes them stable kind of the you know what's the lifespan of

49:00 Microbes are survivors and quite a number of them have the ability to sequester themselves when conditions aren't conducive to them. So if there's no food or the pH is wrong or there's a toxicity or something like that, there's quite a number of different species that have the ability to go into different forms of hibernation. So there's at least four different ways that a microbe can either stop or substantially reduce its level of respiration. And then at that point they become extremely durable.

49:42 To give you an example of how durable they could be, in a core that they took out of a sea bottom into a strata that they knew had been geologically isolated for 23 million years, they were able to pull up microbes. And as soon as those microbes had suitable habitat and food, they woke up after 23 million years. So microbes are survivors and they're extremely durable and they're opportunists because as soon as there is food and habitat they wake up. Not all microbes have that ability but a lot of them do.

50:23 So would that make sense then why in a conventional system of high synthetic use, whether synthetic chemicals or fertilizer, the microbes are still there, maybe they've just gone dormant because of those products being used?

50:43 Yeah, sometimes that's the case. If you had a sub-lethal application of say a fungicide, the fungi might be in a spore form which might not be vulnerable to that level of toxicity. So yeah, you're right, it would just sit there and wait it out until the level was something that it can tolerate, then it would come right back out.

51:11 Yeah, I guess that kind of works into this question from Lisa: how do seed treatments, especially fungicides, interfere with micro colonization and germination? How many days of growth until that seed treatment is diluted with root growth and colonization can then occur?

51:32 Well, good question, and that is so challenging. I know that so many seeds—it's hard to even find naked seeds that haven't been coated with some sort of fungicide. And you're right, it is very counterproductive to us getting that seed colonized. So if you don't have a known high infectious pressure of a particular pathogen you're trying to address by having that fungicide seed coat, if you can avoid it, don't do it. Because all you're doing is suppressing the biology and its ability to assist the plant.

52:12 And as to how long that lasts, I don't know that that's really an answerable question. I don't think it's going to be—it's so situational. It depends on the soil type and moisture content and maybe temperature, and even the community of microbes that are there to potentially colonize. Some will have a lot—actually some won't be bothered at all by it and they might be able to colonize right away. Other ones will never colonize it.

52:46 I think that transitions us into our next question: how does air temperature impact efficiency? Is there a minimum air or soil temperature for these biologicals or microbes in general? And maybe even on the maximum end of that too?

53:05 Yeah, there's microbes. Their metabolism is definitely affected by temperature. So as it gets colder, their metabolism slows down. They're doing less, consuming less food, producing less metabolites, and they'll be less active. We certainly pay a lot of attention to the temperature ranges in which we propagate our biology because we do think it can affect the composition of the community. But microbes—there's a lot of microbes that will be reasonably active all the way down to say 45 degrees. And of course you can freeze them and you don't really lose population, but they certainly become very very slow at that point.

53:53 And then on the upper end of the scale, we certainly won't propagate microbes above about 80 degrees because again it changes the population. If you're applying them, I'd say it's a lot less sensitive to that, especially if you're putting it through irrigation. I wouldn't say there's any upper end temperature cutoff for irrigation applied. If you're fully implying—I definitely don't like to fully imply anything above about 90 degrees.

54:27 If you are applying microbes, make sure you put microbes on with food, so either one of our wettable powder foods or fish, but also something that creates additional habitat and UV cover, so either usually for UV cover a humid kind of some sort.

54:54 We got a question here from Gene. What percentage of the plant nutrition comes from the rhizophagy cycle, the oxidization of the micro shell and stripping out the nutrients? I don't think anybody has that answer. I had the picture of the microbes going into the root hair and that whole process. I don't think that's been documented even. The process has been documented for say more than about five years. In terms of quantifying it, I think it would be different depending on the biology and the variety of the plant, even as much as the species. The GMO plant might be much less conducive to that relationship than say some other species of plant.

55:55 The other type of microbe feeding of the nutrients is those endophytes that I was talking about, the fungi that colonized partially inside the root. They're actually part of the vascular system of the plant at that point and gathering nutrients and transporting them through those hyphae and then delivering them directly into the root. So there's two very different kinds of modes of delivering nutrients and I couldn't even hazard a guess as to the relative proportions between those. I'd say it'd be much more likely that the bacterial function on that root hair would be the thing that you probably see the most commonly. Less commonly the fungal relationship, mostly because there's not that many production ag soils that successfully have that fungal representation.

56:52 I believe we have a YouTube video with Dr. James White that talks about the rhizophagy cycle so maybe there's some more information on that you can get out of that video as well. I love that video. It's so illustrative of it. They've done such great work there and that's what I'm describing is their work.

57:23 Got a question here from Chad. How does electric conductivity of the soil affect microbial population or inoculation? Sure, yeah. The electrical conductivity is a common measure of salts. There are some species of microbes that are pretty salt tolerant and then there's ones that are not. As you build more salts in the soil you start to lose some component of the biology. There are some species that are very salt tolerant and ones that are not.

58:08 That probably goes along with some of the reduction of fertility with high salt nitrogen or phosphorus products. Actually, I'll take that as an opportunity to say something. The synthetic fertilizers tend to disrupt that relationship between the plant and the biology where the plant is feeding the microbes sugars and signalers to collect nutrients for it. Because if you're already putting synthetic fertilizers in ionic form, the plant doesn't really have the motivation to invest energy into feeding the microbes.

58:54 As long as you're not using prilled or particulate solid type fertilizers, those I definitely have a preference if our growers can find ways to migrate away from those products. Because every one of those little particles when it lands on the soil is basically this little salt ball. You've got this gradient from right next to it that almost none of the microbes can tolerate because it would dehydrate them just literally through osmotic pressure, pulling the liquids right through their membranes. As you go away from that in distance you can see more and more biology being able to tolerate that little salt bomb. To the extent that we can use liquid fertilizers instead of those particulate ones, we can do ourselves a lot of favor in terms of the conditions for our biology in the soil.

59:54 Got a question here from Jim. Do protozoa and nematodes populate as well?

1:00:00 With your products, there are some protozoa in some of our products. Those would be the intestate amoeba and the flagellates. Very few if any ciliates and no free-form amoeba. Those are really the recyclers in the soil. And those typically will be pretty well represented in the soil and as soon as you build the rest of the biology, you've created the food source for the protozoa, so those should really re-establish themselves pretty easily. And then in terms of nematodes, we do have one product that has nematodes in it. And of course they do a lot of important nutrient cycling and cleanup work in the soil as well.

1:00:58 Another question here: when applying the products to the residue products, is whether a factor in success of those products breaking down that residue. Metabolism is somewhat dictated by temperatures, but the decomposers do pretty well in cooler temperatures. And that's one of the reasons why if you looked at that part of the crop program, the example one there, there's a little bit of fish on there with them. And as well as food, and what it's doing is kind of giving a jump start of the population to really get deep into those tissues and have a reasonable carbon and nitrogen ratio to get the population rolled, so that'll sustain a higher level of activity through the winter.

1:01:52 I think we have a few more questions here, so we'll continue on if that works for you David.

1:01:59 So is there a good way to test the microbes that are on the seed and how much we'll see growing in your gender system compared to seen grown in a conventional system? You know the microbial difference maybe there. I'll tackle the first part and then maybe we can talk our way through the second one. So if we wanted to quantify and characterize the biology that's on a seed, we could actually do probably take an untreated seed or some seeds and then take some treated seeds and then send them through metagenomics. So if you're not familiar with metagenomics, it's a lab technique that they use to strip the DNA out of the microbes and separate out the viable DNA from the non-viable DNA so you know which ones are actually kind of living biology. Then they sequence that DNA so create this really long string of all the DNA, they pattern match that to a database, and then you can if you get a match, you can name, you know, what's the name of this microbe and what does it do. There's processes you can also do that are quantitative enumeration, so you can actually get a population. So if we wanted to say what's colonized on this, you know, on the seed treatment and then how much of it is there, we do actually have a tool that would do that. And actually I think you probably would be able to get the endogenous micro species as well, so those ones that are on the inside of the seed, which would be pretty cool.

1:03:37 And then the I think the second part to this question or maybe it's a separate question: how much better will a seed grown in regenerative system be than a seed grown in a conventional system? Yeah, I guess in general I would say my expectation would be that if you have a regenerative system where we've built up all this complementary biology, you have so much more opportunity for things to go well. You know, you should have better soil physical and chemical characteristics, you should have more supportive biology and more of it than a conventional system. So for all those reasons I would have high expectations for the regenerative set of conditions to be much more favorable.

1:04:24 I think we have one last question here: what type of soil test was used to get all the results that you showed on the treated versus untreated in the soil case study? The restoring actually, that was metagenomics. So we took some soil that was untreated versus the treated soil, sent both of those to metagenomics, and again it gave us a quantitative enumeration in all the species. And then from there we kind of broke it down as to what their functions were.

1:04:57 Very good. Well David, thank you for giving us your time for today to invite us on into this microbial biological process in the soil and also on some of the products that you carry. And I want to thank everybody else for attending today. This will be again recorded and put on our YouTube page. Next week Wednesday at the same time, we will have Scott shimer and some of the elevated AG team talking about the hyper girl product and what they've seen on I believe Scott's farm. So join us next week, and again David, thank you. Everybody have a great day.

1:05:42 Thanks so much for having me, appreciate it.