 My name is Mike Metzger and the Vice President of Agriculture and Research at Midnight Farms Cooperative and how the pleasure of moderating our mid-warning session. Our first speaker today is Dr. Melissa Wilson from the University of Minnesota at St. Paul and I'm going to talk about liquid-separated area manure in a sugarpine rotation. Dr. Wilson. Thank you, Mike. So they didn't put me after lunch today, but they did put me after break, so I'll get to talk about all of your favorite topics, Menor. We know that we have a lot of some of the larger dairies coming into the sugar beet growing regions, so I figured why not see if the dairy manure is good in a sugar beet rotation. And one of the interesting things about these new dairies is that they're using a, not really a new technology, but new to large dairy technologies, where they are separating the liquids and the solids. The solids actually go back into the barn for bedding and then the liquids are primarily what's applied to the fields. So we're going to talk about our research using that material. So we're doing this at two locations. One is in West Central Minnesota, just west of Wilmer, we call that our Murdoch site. And then our second site is located up near Nashville, Minnesota. And we started our West Central Murdoch site, this is our second year, so we started this in fall of 2019, and then I've had two growing seasons. And we just applied manure in the fall of 2020 at our Nashua site. We had actually intended to get both of these sites started in the same year, but the Nashua site in 2019 was flooded, because if you remember, it was very, very wet that year. So we weren't able to start the trials then. And then here's a photo showing what the liquid separation process looks like at the dairies. This is the solids part that then is then taken back and put into the dairy barns as bedding. So the liquid part is what actually gets sent out to the fields. So our plot set up in our first year is when we applied manure, and then we actually grew all three crops in a rotation, there was sugar beet, soybean, and corn. And we kind of wanted to apply some of the rates that the dairies are offering. They're typically offering a high rate. In this case, it's about 15,000 gallons per acre. It changes from year to year depending on what their nutrient content looks like. And then the low rate is roughly about 10,000 gallons per acre. And then we compared that to a spring fertilizer. For our manure plots, if we needed to add additional fertilizers to balance out the nitrogen needs for the various crops, we did do that. So like our low rates didn't have as much nitrogen applied as the high rate. So we did add fertilizers to kind of offset that. Then in our second and third years, we actually were able to just apply fertilizers. We took a nitrogen credit for our different manure plots. And then we also did soil testing as well for PNK. So we did adjust the PNK fertilizer based on soil tests for the different treatments as well. Here you can see my team is hand applying the manure. This is at our Nashua site. In this case, because we were doing small plots and we wanted to try to be as consistent as possible, we hand applied it. And that basically means we filled up some totes with as much manure as we wanted per plot, lifted it in the air, and gravity fed it down a tube. And they are smiling. I didn't like force them to do that. It was actually quite pleasant because this is the second year we did it. We knew what we were doing. It worked out actually pretty slick. So all the manure then was incorporated within 24 hours. At the Nashua site, we got it incorporated that evening. At the Murdoch site, we actually incorporated it the next morning. Here's an aerial photograph of our Murdoch site. You can see all of the different crops within each row of crops. We have a corn, soybean, or sugar beet. And then we replicated those four times. So to keep things short, I'm mostly going to show a few photos and then get right into some of the yield data. This is July 2021. So our Murdoch site was in its second year of the rotation. I presented on the first year at this presentation session last year. If you remember, in our first year, we had some IDC, iron deficiency chlorosis issues with our soybean on our manure plots. And this particular year, we saw some of that, although it was a lot more random across our field. Though we did see one in particular where the low rate of manure seemed to affect the soybean more greatly than other places. So we were still seeing some of those iron deficiency chlorosis issues in soybean. Our sugar beets looked really good. Our corn looked really good. Remember that this is the second year. So the fertilizer was applied across all the plots. We did vary the fertilizer based on manure, nitrogen credits, and soil test P and K. Nashua, this is taking the same day earlier in the day. Again, this is after the first year of manure application. So our corn looked really good. Soybean this year at this location looked really good. It turns out the pH was a lot lower at this site. It was about 7.3 versus it was about eight at our Murdoch site. And we can tend to see more iron deficiency chlorosis issues in soybean at high pH, especially when you add organic matter. So we didn't seem to have that problem. We also chose a better soybean that had more iron deficiency chlorosis resistance too. We did see some stand issues with sugar beets. I think we found out like a day or two before planting that Callisto had been sprayed on this field the year before for the corn. So we anticipated that there might be some carryover residual issues. So we did plant it a bit of a higher rate. And because it was so dry, we definitely saw some of those issues early in the season. It does seem like it kind of outgrew those at some point though. So getting right into yield, we'll have our corn yield here on the left and our soybean yield on the right. This is year two after manure application. And our fertilizer yields. Well, first, I have to mention that our corn yield ended up lodging right before harvest. So we had to go in and hand harvest this. So this is all hand picked green. And our yields ended up being 157 bushels per acre where fertilizer was applied. With our low manure rate, it was about 10 bushels per acre higher at 167. And our manure high rate was our best yield at 185 bushels per acre. So looking at the statistical analysis up here, I have letters and any of the bars that have a different letter above it are statistically different. So for our fertilizer plots, the only statistical difference was that manure high rate. That was statistically higher than the fertilizer only plots. A lot of potential reasons for that one manure has a lot of nutrients besides what we would apply in fertilizer, has micronutrients, has some of the secondary nutrients that we hadn't necessarily applied. So we think that those are some of the reasons why we were seeing these yield differences. With our soybean, there's error bars here you can see that are really large. And that's because of some of the IDC issues that we saw randomly across the plots. So we didn't see any statistical differences in yield. But we think that's mostly because it was just really variable across the site this year. Our first year, our fertilizer plots yielded a lot better. I think it was about 40 bushels per acre and then we had like 13 and 8 bushels per acre. It was really abysmal where the manure had been applied. In our second year, a lot of the treatments were very similar. But again, high variability. Here's our data on sugar beets. We have a table because I have yield and this is root yield, extractable sugar in pounds per ton, extractable sugar in pounds per acre and sugar purity. And then we have fertilizer only in the top row, low dairy manure rate in the middle row and the high dairy rate in the third row. And again, remember, because this is Murdoch, this is our second year after manure application site. So overall, our yields were very similar across all treatments at about 40 tons per acre. Our extractable sugar, again, very similar across all of our treatments, averaging about 271 pounds per ton. Just see a little bit of a difference in extractable sugar in pounds per acre. But overall, statistically, they're all similar. They're from 10,844 pounds per acre up to 11,051. So not giant differences. And our sugar purity was interesting this year. The fertilizer only was about 92.2%. And both of our dairy manores were in the 91%. But again, we didn't see any statistical differences across this. So we did see a little bit of a decrease in purity. And we did see that that first year as well with our manure plots. But we do tend to get similar or higher total sugar per acre. So at Nashua, this is where manure had been applied the previous fall. Our fertilizer yielded really well 230 bushels per acre. Our manure low rate actually yielded a little bit less at 209. And our manure high rate was 228 bushels per acre. Again, statistically, we didn't find any differences. But the high manure rate and the fertilizer were all very similar to one another. Interestingly, the soybean yield were quite different than what we had experienced at Murdoch. Where we applied manure, we actually had tended to have better yields. Again, there was not a statistical difference, because there was a lot of variability. But we tended to have 56, 57 bushels per acre compared to the fertilizer only plots, which were only about 49. And then here we have our sugar beet yields for the first year after manure application. Again, we have nutrient source yield, which is root yield, extractable sugar and pounds per ton, extractable sugar and pounds per acre and percent purity across the top. And at this site, we were pretty pleased with the yields, actually, because again, we had seen some of those stand issues early on in the season. But our high dairy manure rate actually had the highest yield or tended to have the highest yield at 41 tons per acre. And our low dairy was actually the lowest compared to the fertilizer. Our extractable sugar was a little bit lower with the high rate of dairy manure at 271 pounds per ton versus the low dairy. The low dairy and the fertilizers were about 282 or so. Our extractable sugar was all very consistent across all the plots. However, ranging from 10,700 pounds per acre up to 11,200. So not a huge range in sugar. And percent purity, what's interesting here is the low dairy manure rate actually had slightly better percent purity than our fertilizer only plots. But again, it wasn't statistically different. They're both about 91%. And then our high dairy rate manure, we did see a little bit of a hit at 90.8. So overall, the yields and how this factors out is gonna kind of depend on if the percent purity really factors heavily in your payment system, then you might see differences than if it doesn't. So I wanted to keep it short and sweet and it looks like we're running a little late. So hopefully we should have plenty of time for questions. I'd like to thank the Sugar Beet Research and Education Board for funding. I'd like to thank the MnDAC growers as well as the Southern MnGrowers for helping with these research plots. They're amazing teams and have been great. And then we also had some custom people come in and harvest for us. So thanks to those groups as well. Follow me on Twitter. And I do have an update that I'll be kind of going through the next few weeks and updating it at z.umn.edu slash manure sugar beet rotation 2021. Hopefully include a lot more details in there that I wasn't able to cover here today. So make sure to check it out. Very good. All right. Thank you guys. Okay. Our next talk is sensitivity of CLS, the Full Year Fundersides in 2021. The talk was prepared by Gary Sikor, Viviana Rivera and Dr. Melvin Bolton. Many of you may not know, Gary was actually my advisor for many years during my graduate school. And like any good mentor, his influence still holds sway many, many years afterwards. And he had a personal commitment today. Apologize us for not being able to be here. And I will do the best I can in his stead. This last year, Gary and his team did 592 samples, circus per samples. This is just shy, just half shy of what they consider normal for any year. And the reason for it, it was the COVID supply chain, specifically the specialized plastics. They could not get petri dishes, pipette tips, all these other kind of specialized things to do what they did. So they asked all the cooperatives to cut her back and scale back a bit. What you're looking at here on the graph is looking at field incidents in the blue bars, which is the number of fields with resistant isolates present within their borders. And then the sport germination of those isolates. This is population-based data. Gary always has said there's a fitness penalty with tin. So what that means is when their tin is not used, like you see in the middle here, right around that 2005 and 2007 era, there's a lower incidence and a lower percentage of spores present. When it's used highly, like it has been in 2016 forward, you can see it ramping up there. That's because it has been really a staple in all the fungicide programs and all of the cooperatives. That's the reason for the increase. What will be interesting going forward is to see where this lines out with the introduction of the new CR plus varieties. Will they require a lower number of sprays and therefore a lower use of super tin. So given the date on the last slide, it's no surprise that the level of tin resistant increased in each factory district. The most significant jump came at Southern Minnesota, or excuse me, in the Crookson district, Southern Minnesota got the silver medal and Mindak the bronze medal for tin resistance. So switching over to the DMIs now. The way Gary has explained DMIs is he uses something called the resistance factor. The resistance factor is the EC50 value divided by the baseline sensitivity. He does this in order to make comparisons of population shifts from season to season and compare between them. The EC50 value is the concentration of a fungicide that inhibits mycelium growth or spore germination of 50% of that population. The baseline sensitivity then are how sensitive the isolates from 1997 and 1998 are to the same fungicides. Again, that divide the two out, that's where you get the factor. Now, while all the DMIs have resistant factors around their eyes, proline still continues to be one of the better ones and stands out from the others. So when we look at these individually, you can see there again, proline, so the lower the bars, the better off you are here. Proline continues to be the best followed by eminent. It's interesting with eminent given, you would think that given its low level of use up and on the valley over the last few seasons. What is interesting is inspire and provisal two of the more popular ones are on the highest level of increase. It's also interesting that they share a very similar pattern because it has been shown that they exhibit cross resistance between each other. So what's good for the goose is good for the gander in that case. Looking at the percent of isolates with eminent resistance across the factory districts. Now the way Gary does this is he uses a 10-fold scale over here. The higher the level of resistance, you're the more colorful the bar, the more resistant that particular AI is to a specific fungicide, in this case eminent. So it's virtually the same across all factory districts but it is encouraging that there are very few isolates with what he would consider 100% resistant to this chemistry. So what about inspire? Little more red across the board with southern men in Mindak showing the highest levels of resistance to inspire this past season. Proline, there's a lot more red or again resistance when evaluating proline. However, the jury is still out on this data set is not that they can only test sensitivity to what's called pro-phioconazole, that's the active ingredient in proline and they can only do that in vitro. And in other plant systems, there is a desthiometabolite that is produced that does the heavy lifting at the fungal target site where inhibits the production of agrosterol. So the $64,000 question is, are they testing the right component, pro-phioconazole or the desthiometabolite? And the jury is still out on that, like I said. However, there's a fellow named Dr. Nathan Wyatt at the USDA in Fargo. He's gonna look into if sugar beets are able to produce the same phenomenon. So stay tuned on that. Percent of isolates with provisal resistance. Again, this looks eerily close to the inspire data set but given the cross resistance between the two AIs, it doesn't come as much as a surprise to anybody. And the way Gary explains it, DMIs are like pistons on an engine. When one is high, the other seems to be low and eventually we start using that one and then the other one shift. And you kind of go in this back and forth pattern and everyone eventually takes their turn on top. So what about headline, if I can get it up here? All right, headline in the QOIs. Basically same story, 10th verse. Now this is population data similar to 10. However, the differences is done with a molecular test. So it's done on more spores and it's done in a more accurate fashion, one could argue. What you're seeing on the bar on the far right there is that 95% of the isolates tested this last season had at least a 50-50 blend of resistant isolates with the G143A mutation present or higher than that. So a lot of resistance out there. What they are working on is trying to get this same type of molecular technology transitioned into the DMIs. They're just not quite there yet. How they do it now is they put technical grade funericide into the auger, sterilize the auger, put a single spore on it and then let that spore grow out for three to four weeks before they take the rating. It's a very labor intensive and very timely process, time and long process. So what the Dr. Bolton lab at the USDA has done, he's identified an L144F mutation. This works on eminent and they can directly correlate through PCR work comparing the EC50 values to the PCR CT values and come up with how resistant that funericide is or is not or the spore is to that funericide. However, what works on eminent does not work or appear to work as well in some of the other chemistries. So they're only to get a 60 to 70% correlation amongst the DMIs. That's where it is today. And Gary must have been watching Anchorman on Netflix because he quote, 60% of the time it works every time. So sensitivity, again, the headlight looking across the factory districts. Now what's interesting here, MNDAC has been one of the lower ones since 2016. And that's when we stopped using headline for sarcasm or control there. So in a sense, over the last five years, we've really kind of been evolving them, those spores out of the population. But when you take a look for between 2020 and then 2020-21, this is the first time since 2016 that we've actually seen an increase in that funericide sensitivity to headline at MNDAC. Now at MNDAC, we've been preaching to our growers to spray early. And Joel Hastings is in here somewhere. He has the best quote. If you think it's too early to spray for sarcasm, you're probably right on time. And he's exactly right with that. In spraying early, we've preached to the growers. It's the practice of spraying early, not specifically the product. And this last season, we had a few guys that had headline, some Praxer, in the shop yet that they wanted to go through and they went out and applied that for the very first funericide application for plant health benefits and some other things that have been documented with that. We need to follow up on this, but this is one hypothesis as to why we are seeing that increase and we haven't seen it before now that that funericide was being used again. So if that is the case, it shows how fluid this population could be even with the introduction of a funericide. So again, stay tuned on that. Now these are some slides that Gary asked me to add. At MNDAC last year, we were about 60% of our acres were planted to the new KWSCR plus trade. We had a lot of success with it. It was a very good thing. When we went and sampled and submitted our samples this year, half of the samples that we submitted, it's a 48 of them taken by the staff were CR plus, half of them were what we would call the traditional varieties. And then indicated as such and Gary ran them. What's interesting is that on the headline here, so this is the bar on the two on the left, CR plus on the left, traditional on the right with the multicolored bars. The CR plus 84% of the isolates there still had levels of that greater than 50% of the spores were had the G143 mutation present. And on the right, 89 of them did. So really not a lot of difference there from an initial look at it. Tin almost identical, 64 and 64 across the board. So what about the triazoles? Now the comparisons you can make here, the CR plus varieties are all the bars on the left, the traditional varieties on the right. And if you wanna make comparisons, for example, taking a look at eminent each one, compare the 11.9 bar to the 8.7 bar on their traditionals. When looking at this, and again, this is just a quick first glance, there's several patterns that develop in the dataset. And this really speaks to the consistency of the work that Gary and his team do. You can see the eminent increases on the CR plus, the eminent increases all the way up to the greater than 50% of ice is sampled having some resistance present. You can see the exact same thing on the traditional. Look at the provisal, low, high, medium, and then high, the exact same thing. So when looking at this and evaluating it, there's two key things to note. One is that nothing really jumps out. And that's a very good thing. It does not appear that using these type of genetics that we are selecting for or promoting resistance to any specific AI. That's a very good thing. And then the other thing is while these genetics stay very clean and stay very green long into the season, the spots that do develop on the CR plus varieties can harbor spores that have a level of funder side resistance. And therefore it really hammers home the need that under no circumstances should a single mode of action be applied. You have to tank mix every time. One to protect the chemistry or one to protect the chemistry and two to protect the resistance that's built in there. So they have to have a tank mix partner, absolutely no exceptions. So to kind of wrap it up here, Gary has tin as our best weapon, something only the US has. It is increasing in both incidence and levels of resistance, but that's likely due to it being in rotation with the triazole, tin triazole, tin triazole back and forth. It'll be very interesting to see what the CR plus varieties have in store for us. Topsin, they did not test Topsin in 2021. And that's because they know the context of the book before they even read it. The last time they did 98% of the isolates had the mutation to Topsin present. And that's something that doesn't go away. That's present ever present in the population. So short answer there, don't use Topsin, period. Triazoles, this is where Gary says the action is. The EC50 values have increased for proline, spire, provosal, but not for eminent. Again, theoretically lack of use, if you will. The resistance factor increases for eminent, inspire proline, or excuse me, provosal, but not proline since 2017. He hopes there is a fitness penalty. They're seeing if they can evaluate that. I talked about the transition into molecular testing for this. And then always apply it with an EBDC or copper partner. That's kind of the message there. With the QOIs, there's lots of the G143 mutation present out there, a greater than 90% since 2016. I spoke a little bit about the MINDAC. We'll follow up with that and get that information to Gary and Viv. And then again, his recommendation is that QOIs should not be used for CLS management. Still be used for frost protection and the plant health, absolutely. But specific for CLS control, that's just not the best idea. So utilize the stuff that your agriculturalists have been preaching to you from resistant varieties to the cultural practices, and then early funderside applications with TankMix. That's gonna be kind of the take home message. Gary made sure to acknowledge the SugarBeat R&E board for funding of this work. He thanks the member companies here for supplying the technical grade material for use in this experiment. And of course, Judith, Viv, and the whole team of elves as it tells them doing the work. So with that, thank you very much. And next on the list is Dr. Chu. Improving SugarBeat resistance to stochastic relief spot through genome-wide association studies and double haploid. Dr. Chu, he is with the USDA in Fargo, North Dakota. Okay, hi, I just want to talk about improving the SugarBeat resistance to stochastic relief spot through genome-wide association mapping and double haploid. So stochastic is a very severe disease for worldwide. So it's the year of the losses we are known as high as 42 to 50%. And for the host resistance, it can be controlled by many genes or like recently, this is the CR plus assuming like a major gene. So we use the genome-wide association analysis. So in this way, we can find the association between the markers and the resistance. So right now, mostly we are using the SNP markers as a single nucleotide polymorphism markers. And using association mapping, we do not need to make the specific mapping population. You can just use the natural gene-plastic connection and also use the association mapping. We can increase that's the mapping resolution because what we detect is the recombination was occurred many, many years ago. That's all called historical recombination. So once we identified the resistance through this association mapping, and then how we can to apply the resistance to develop the resistant gene-plasm. And using double haploid, it could be a very efficient way to develop this new resistant gene-plasm. So here just let you know a few of the concepts. This is the diploid, that's the regular sugar beads. We are growing this. So that has two sets of the chromosome in each cell. And the haploid of course, that's when we have of the chromosome from the diploid. So that's each cell only contact one sets of chromosome. And then we can use this chemicals like a cutscene to double the chromosome number to make this double haploid. So the double haploid actually is based on the haploid. We just double the chromosome number. And so make this back to the diploid. So if the haploid is not useful for breeding because it cannot get a seeds, it was only one sets of chromosome. So we have to make to the double haploid. So make it to the diploid that can make the seeds. And so the double haploid compared to this diploid, so that's the two sets of chromosome that's the double haploid was identical. So from genetics, we say this is completely homozygous. So that's when you grow there, you never get this trigger segregation. So all we can say is genetically very stable. And the most benefit using double haploid is to make this the breeding process much shorter. So here I just give you this as a diagram to show to compare this conventional breeding methods versus DH technique. So if we use a conventional breeding method, you will want to make the final as more homozygous lines. We need at least six generations self-pollination. So it's non-process. I mean, it's pretty for sugar beet. If one generation is more than one year or two year, then we have over 10 years we can develop a germ plasm. But if you use the double haploid method, so we use this tissue culture, we induce this the eggs from each flower and then we can directly using chromium doubling and to make this double haploid. So these only need about one year. And also the home homozygosity is 100%. And with this traditional way, we cannot get like 100% homozygosity. So this was, we can make this process much faster. And in our idea, we want to start from a big connection of this germ plasm. And this including the sugar beet and other cultivated the beet and also the wild resources. And then we do this trait analysis for customer evaluation in the field. And then we use this generic data mostly snips covering the whole genome to do the association analysis. And then from here, we can select the parent lines, particularly for the ones that have very good resistance. We make the populations. And from device, we also can identify the markers associated with the resistance. And so in that way, we can use that to develop markers for marker system selection. And then we can use the double haploid method, use markers to select the station lines and confirm with disease evaluation. So in that way, we can make this new germ plasm with improved cost per resistance. So our objectives, we want first identify of germ plasm lines that had showed the resistance to sarcospera. And then we can use the GWAS to identify the genes, confirm the resistance. Then we can, we use the double haploid to develop this new DH lines with sarcospera resistance. So totally we have almost 2000 germ plasm accessions were collected. And so this majority was the, this bad maritima, this is a wild ancestor of the cartilage beads. And then we have over 1300, this is the cartilage beads, mostly including is the four other beads, veggie beads, naquif veggie and root veggie. And over 1000 sugar beads, all these. And we have some other, this is a wild relatives. And from here, we get the genetic data from 1928 lines, that's only eight lines, we didn't get the good marker data. And overall, all this marker data was very good that we're using the reference genome. So that's the marker was covering the whole genome according to the reference genome sequence. And we did this population structure analysis to all those accessions. We found is the, it has five subpopulations, but when we look at here, like the five sections, only the first one, mostly kind of the color more red. And then from this part, mostly sugar beads, this is maritima there and all this cartilage and maritima another section. So you can see here has more colors mixed together. So that means all those lines in these sections was more too close. So that's like directly all mixed together. Then we use this another software to show with all these clusters. And we can see the here, this was one cluster. Then this is the second class, third, this is the fourth and this five was very big, huge one. So mostly this cartilage, the sugar beads and the other type of the beads. And, but the good thing is we're very happy to see the first cluster. So you can see here has more distance to the rest of them. So that means this cluster was genetically more distinct from this cartilage sugar bead. And so the good thing is if the genetically more distinct we can use this to broaden its genetic base of the sugar bead. So in that way can increase the sugar bead sustainability for production. And when we look at this section of this, it mostly is from Bithmeritima. And it was collect mostly in France, England, Ireland and the Morocco. So mostly was in this reading and here. So, and then we, I look, went to this the National Plant Jumping System, the stat base. We found some lines already have been evaluated by other researchers and see what's your reaction to the sarcospera. And I find the 797 accessions from my collection was has data available. And here this briefly was the reaction to the disease. So they use this zero to nine reading scale, like a zero to three, zero is immune, one to three was the resistance. And so from here, I can see that Meritima has average reaction was lower than this cartwheat speeds. And when we look from this heat scrum, we can see that the Meritima here has more resistance but for the cartwheat speeds, so much less resistance there. So that's good. That means Meritima could be a very promising this is a resistance source for sugar bead improvement. And then I compare to this with, I just curious to see how many lines in this cluster as this Meritima has good over. Then we find this 198 Meritima accession has data available and 44 as rate is one. So that means very resistance and 25, 27 was two or three. So that means you have pretty good resistance there. And then we still have another 160 lines. We have no data available and we can evaluate. Maybe we can find some good results from that team. So just use this historic data. We run this genotype, this is non-white association mapping and we find some locus was more associates with the resistance. You can see many this Chromadon one, this Chromadon four, five, six, seven, eight, nine. So and to my surprise, I have thought of this data was at the beginning I thought that maybe not very good but from the QQ plot, I can see this line means what's the expected but this line here was means observed. So if this more towards the observed, so that means this data was more positive, good. So, but anyway, since this is a historic data that's connected from different, maybe different environments from the different time. So we definitely still need to confirm this results is the new experiment. And then I talk about what's the double haploid system what we are working on. So we use this unfertilized flower and we get to the all wheel and do the culture we induce the callus and then we use this callus to get the seedlings and we plant the seedlings in this soil and then we cut this root chips to the chromosome counting and we find this each cell only carry nine chromosomes. That's a half from the number of this diploid. So diploid is 18 chromosomes. So that means they're haploid. So then we will use this callus tissue or use this haploid seedlings to do the constant treatment for chromosome doubling to make the double haploid. And we start to do this double haploid in January, 2021. So we spend about half a year. So we cut it up about 5,000 all wheels and we successfully get the callus tissue from this 27 individual all wheels. So we obtained a lot of haploid seedlings. So right now we're working on this chromosome doubling. So just quick conclusion and from what we learned, so that in this collection, the genetic base of the sugar beet is narrow. This already indicated from previous reports, but we found this subset of this bad maritima lines could be important for sugar beet not only for this improving the resistance, but also for broadening the genetic base. And we find some of this association between some not reaching with the resistance and from this historic data, but this will be needed to confirm and in our new experiment. So we already have this double haploid system developed. So we are testing that use more genotype to increase this efficiency. And also thanks to my team members and also my colleagues in USDA, ARS and also the NBSU and the University of Minnesota and also thanks to BSDF, USDA, ARS and should be the research and the catching bot for this funding spot. And thank you all. The next topic is going to be factors affecting the CLS for germination. This talk is presented by Ms. Villanova-Harris and co-authored by Dr. Yuri Seaport of Closed State. This is the second part of the study we did last year. So thank you. So this is our first one to do as Maria or what we found last year. So far what we know about the condition for export termination, we know that free water is more important than high level of humidity for termination. Germination occur at lower temperature that previos assume. We find out that temperature as low as 10 Celsius or 50 Fahrenheit, we can see a start of the germination process. The germination process start about two hour of incubation in free water. Germination increases as the temperature and incubation time increases. Export production and germination may occur earlier on resistant isolate than the sensitive one at temperatures of 20 Celsius or about 62. Just kidding, sorry. So this is the summary slide for the project that we need last season. So we can see that the free water across all the treatment and isolate, we see that it's a 44% termination against 2% termination in 100% relative humidity. Now if we look the temperature, we see that it's a steady increment in the percentage of germination as the temperature increase. Now if we look the time for hours that incubating, we see the effect of the time of incubation is higher in the different hours from four to eight, 12 and 24 hours. Also we found that isolate, resistant isolate in general have less termination, mostly in lower temperature that the sensitive isolate. So here is an example of the relative humidity at 100% and compared with the free water where we have a lot of termination and produce some mycelium from the spore. And also in the left corner, we can see the start of the termination process at the tip of the spore. Another thing that we observe that in lab condition, when you have resistant isolate and sensitive isolate in those spore production, we saw that resistant isolate produce more score or faster the score that the sensitive water. So based on this information, we was looking to see what is the impact on interrupting this wet condition or wet incubation period on a spore germination. For this, we conduct this study in laboratory condition about 65 Fahrenheit. We use five spores that were resistant to the DMI site UIs and this isolate were the same isolate we used in the previous study. The spore production was induced for all this isolate when the spore was ready, we collected with the sterile distilled water and we placed an aliquot about 100 micro liters in a microcobo slide. And a set of 20 slides for isolate were prepared. So this slide were put in a box with lining with wet paper towel and incubate all this spore for three of two, four, six and eight consecutive hours of wet conditions. At the end of each period, we air dry all this slide. So it took about 15 to 20 minutes to evaporate all the water. One slide was used to determine the baseline germination for each wet period. And the other slide were kept in dry condition for a period of eight, four, eight, 12 and 24 hours. At the end of each of this period, water was added to the spore again and incubated for 24 hours. At the end of the incubation period, germination was stopped using cotton blue and we assessed the percent of germination for each treatment. So this is the baseline for spore germination. This is just right after the wet period. So we can see in two hours of wet period, we have a 16% germination and when we have it in four hours, they jump as almost 63% germination. So that is increased about 46% point in the amount of germination. Now, if we compare with the six hour, they're still increasing percent of germination, but this difference is less, it's about 24 points. And if we go to the eight, compared is about the 6% of germination. So also we found out that it's a lot different between isolate and this graph is each isolate, is percent of germination in each isolate and a different incubation period. We can see in the two hours in the blue bar that we have a germination as low as 9% all the way to 23%, 24%. When we move to four hours, the germination increment from 33% the lowest in the germination all the way to 75%. It's a big jump, big difference in between isolate. When we incubate for six hours, we still have different sports germination, but that different is not as hard, high as in the four hours. Same occur when we incubate it for eight hours. So now if we look at what happened when this wet condition are interrupted, we see the first graph zero is mean that wasn't exposed to dry period. We can see at four hours after drying there is an impact in the sport germination, but at two hours we have 14% of, this is treatment wet, but it's for four hours to dryness. We can see there is a different sport germination. But if we move to eight hours of dryness, we see a two hours of incubation have big difference with the other four state 12. And in between then there is still different but it's not as pronounced. And the same happened when we keep drying this sport for 12 and 24 hours. So as a summary, using all the information and trying to get sense of this, we can see that in the isolate there is a significant difference between the percentage of germination in the different conditions. But also see in the wet period, we have average of 16% in the two hours incubation and a dramatic increase when we have four hours of incubation. And this increase is a steady but it's not as marked or as shown as the previous incubation. And also in the dry period we see difference in the sport germination but this difference doesn't have as much impact in the sport germination as the wet conditions. So as a summary of this, we can see free water is really important for the sport germination of Cercospora veticula. Germination increases as the wet period increases. Dry period times after wet condition do not affect a continuous sport germination as much as the wet period does. This data suggests that intermittent wet and dry condition do not have a big impact in the sport germination. There is a significant difference in sport germination among isolate after a various wet and dry period. Four hours of the continuous wellness have a big impact in the sport germination. And there is a study that in 2021 and then in 2000 that used the weather data. And also they showed that four hours in rainy condition were the most conducive for developing of the Cercospora veticula, Cercopora leaf sport. So this is two study help us understand what is the characteristics of the condition that is for need for germination. So we want to use this information to try to develop an early force test model using weather data and also presence of the sport in the field. So for this, we proposed this study doing early detection of Cercospora veticula, sport production and also detection of infection in commercial sugar bits field. For early detection, we use a sport trap called Espornado and weather station that were installed in six sugar bit fields. Weather station also were equipped with leaf wellness sensor as early work in our lab show the importance of free water for sport germination. All this study was done during the May, early May to in the first week of August. The Espornado and the weather station were placed in the field right after planting time. Espornado filter or the cartridge were collected three times a week and tested for the presence of Cercospora veticula using PCR technique. All this study was possible by the collaboration of American Crystal Sugar Company and Southern Minnesota Dictionary Cooperative. Prior to this study, these two companies did an early study, an early infection study in which they sample about 57 commercial fields where they collect the Espornado and assay for the presence of Civeticula DNA using PCR. They did this study from June, early June to late July 2020. And this site also were monitored for the presence of CLS spot. So the result of this study will be presented by Dr. Nathan Wyatt, that many of you know, he joined the sugar beet and potato research unit as USDA in fireball. That's it. Thank you very much. All right, our last speaker before a quick awards ceremony before lunch is Dr. Mohamed Khan. He's evaluating further science for controlling Cercospora v-squad and sugar beet with different levels of susceptibility to Cercospora. Thank you, Dr. Mitzger. Good morning. Good morning. Come on, let's get going. First of all, let's go with Bison if you can. NDSU at cloud nine. Some of you back Alabama, they only have seven. Guess how many NDSU have? You still have one more to go for the next, for the tongue. What do we have that's kind of unique to our sugar beet growers? Good players, good coaches, great team as a result to get great results and phenomenal impact. Work over $5 billion to the state of North Dakota and Minnesota. That's what you are. Our producers are good. Our researchers are even better. I've given you really good recommendations. You use it and what do we have? A vibrant industry and life will be sweeter if it were not for my one nemesis, Cercospora beticula. This came, of course, from Europe. Most of you have European ancestors. They knew about sugar beet way back in the old days. And when they came to the USA, they wanted to feel like home. They brought sugar beet. And guess what? They also brought Cercospora in spot. This disease, we have used funding sites for a number of years for controlling it, but it doesn't matter if you're in Austria, Serbia, any part of Europe. This disease has become much more damaging. And the reason is this fungus produces large number of spores, a trillion spores for acre. And this is what they do. They reproduce, reproduce all the time, take note Americans. We need increased growth rate just like our fungus. And what happens? We cannot control it after a time. Is this only restricted to Europe? No. We collected samples from all over Europe. What we found, they were resistant to the QOI and the triazoles. Likewise, in the US of A, it came a little bit later, but just as devastating. The funding sites which were very, very effective at one time, things like headline and proline and inspire, they became ineffective because of resistance issue. So I will share with you some of the highlights from our research for work done at FOXOM in 2021. You can see in 2021, the site there had some green plots, which meant that some of the treatments were very effective. To the very right, you'll see a non-treated check and that's what it looked like towards the end of September. What were some of the treatments that were effective? The treatments that were effective were those multi-site funding sites. The gold old ones, such as Tin, Mancuseb, Copper. Five applications at about four PDAs interval provided good to excellent control. Now beware, don't go and do this in the field. This is what researcher do to kind of tell us which ones are working well. So we know Tin is working well, Mancuseb and EBDC and a Copper. When we look at the triazoles, they are not all equal. The most effective one was Proline, followed by Inspire, followed by Minerva. And as you saw earlier, we have some other newer triazoles. Unfortunately, in terms of disease severity and in terms of yield, they were not significantly different from the check when used alone multiple times. So what do we recommend? Or when you look at the individual funding sites, what do we see? Numerical wise, in non-finished check, you would expect lower yield. And we see highly spot and lower yield compared to where we use individual funding sites. In these treatments, you had anywhere from 10,000 to 12,000 comes to come of sucrose or raker. We usually recommend funding site mixtures. Where do we get that information from? Right at the resort site. Look at the pictures, they tell the story. For those of you who are new to Beats, we have six row plots, we spray the innermost four and we harvest the innermost two. You can see the innermost fours are looking good with those mixture of funding sites. What do we normally recommend? Different modes of action. An EBDC with a DMI, a tin with a DMI, and other DMI in the form of Proline and Manzi. And when you compare those to the check, you can see you have good, excellent control. Numeric wise again, the non-treated check was significantly lower in terms of recovers sucrose. The least spot rating was close to 10. And if you look carefully at this graph here, the line graph I show you, when you mix the funding sites, your disease severity is even lower when you compare it to individual funding sites. So the way to go is use mixture of funding sites. Now, not everything was rosy. There were some treatments that were not effective. I won't dwell too much on those just to let you know that it's not every product that is out there is effective, especially when used by itself. These are just some pictures early in the season by the end of the season, visually and ill-wise, they were not different from the non-treated check. So what do we recommend to you? We recommend that we use mixtures in a rotation program. So this is what this table is showing us here. When we had an untreated check, of course, the least spot rating was 10 and the recovers sucrose was over 8,000 tons. And in some years, we'll happily take that. But in a year like 2021, where the sky was the limit, using five, six or seven applications in mixtures, different modes of action, what happened? The least spot rating, most of them were less than four or 4.3. And look at the recovers sucrose, over 13,000 pounds of recovers sucrose per acre. When you do the calculation based on American crystal payment scale, you are getting between $2,100 to $2,300 more per acre after you will have paid for your funding sites and the funding site application. So make it while the sunshine, make lots of sugar now so you can have lots of money for the bad years. These are some pictures from those rotation studies. To the left, you had six applications. To the right, five applications. The difference was where we had six applications. We started when we called time Z, which was early, June the 28th at picture on the right. We started a little bit later, July 17th. Now, let me go back to this previous picture here. We were using about five different modes of action. Inspire, a DMI, Manzate and EBDC, Chin, a triphenyl chain hydroxide, Topsin, a benzimidazole, and Priaxor, and SDHI. So we were using different modes of action. In this picture here, if you're from the North, especially and you do not want to use any of the triazoles or any of the QOIs, as Minda did some years ago, if you want to get rid of those isolates, one of the strategies is use the multi-site funding sites to get rid of those resistant isolates. And maybe over time, we can use back those QOIs and DMI's much more effectively. This picture here shows, if you just use what we call the multi-site, you can still have protective control once you go in a timely manner and you're mixing different modes of action. Now, in all of those treatments, we were using 17 gallons of water and 60 PSI in our application. And there were some questions. Should we use less water? Some growers, if you have large acreage and you're far away from your water source, you will spend less time by using lower water volumes. So we just did a study with 5, 10, 15, 20 gallons of water as well as 25. We use funding site mixtures for applications. And as you can see this picture here, we also use three nozzle type. The five gallons per water, each one of these treatment had the exact same amount of active ingredient of funding site. So the amount of funding site was the same. However, the coverage was not the same. And the five gallons per acre was resulted in significantly lower yield compared to the other treatments. So if you look as you went from five to 10, it was better, 15 better and 20 better yet. Dr. Metzger asked, water said it was cheap, but this I'd say is relatively inexpensive. What about if you go to 25 gallons per water? So we did 25 gallons for water and visually that those were the best treatments and numerically the highest cost to close for acre. We like to do some more study to see if this is consistent over time. Now that was 2021, a phenomenal year, more or less dropped early in the season. 2020 it was very wet. And if you look at that picture there, the only thing that was green were varieties that were more or less very tolerant to leaf spot or CR plus varieties. So we did some work using four different varieties. Two of them were very susceptible. A and B, C was tolerant and D was a CR plus variety developed by kid of yes, where they use beta maritima as one of the parent in the background so you can have this resistance. And we use in some of the treatments and I'll show you a little video, five applications each being similar or less applications two or one and we compared it to the check. And in some instances like variety A and B, if you don't use a funding site, you get significantly reduced recoverable sucrose in some other varieties depending on the year, it may not be so impactful. So this picture here, there's a picture that I'm gonna show. The first three treatments have five applications. So you'll see block one, two and three, five applications, block four and five, two and one application. The one that hardly has leaves is a check and then the same thing happens again. Three plots with five applications, then two applications, one application and a check. Another variety, a tolerant variety or more tolerant variety, three treatments with five applications, two application, the one application is not very good, a check and now the most resistant variety with or without funding sites. The last plot on the right is a different treatment. So you cannot see any difference between the check and those that receive five treatments with this particular CR plus variety. Now this is an experimental variety. So we went to the co-ops and the C company and asked for two commercially approved varieties, CR plus varieties to see if we can understand how Cercosper affect these plots. The check, these were inoculated, the check are on the extreme right for two varieties and the picture to the left and the picture to the middle, they had different applications. We could see anything differently, yield wise and least spot control wise. These varieties had significantly high yield compared to anything we've had before. So they were close to 15,000 pounds per acre. We still have to learn how to manage these new varieties in terms of controlling it with funding sites and when to apply it. Now those were very brief. How severe was the least spot? These are pictures from non-inoculated check. This is how severe the disease was at that particular site. So I will conclude by saying, use a holistic system to manage least spot, crop rotation, incorporate debris, use funding site mixtures, use improved varieties. We know our population is still resistant to the QOI, they're becoming more and more tolerant or resistant to the DMI. If we use mixtures, if we start in time in fashion and now that we have these improved varieties. So even if you have a rainfall and the funding sites are washed off, these new genetics will still be able to give us a crop to process and survive. Thank you and thanks to all the people who have made this resource work. R&E Board, seed and chemical company, look for taking pictures, Mike Menzger for arranging with Kevin to give me the best land he has and my students for doing all the hard work. Thank you, Mohamed, very much. You're up. All right, I'd like to introduce Mr. Eric Erdman from Crookston District, I believe. Crookston, yep. He is the chair of our R&E Board and we'll just have a few closing remarks before lunch. Okay, so we have a few awards and scholarships to give out this morning. And we'll first start with the Dr. Dexter Scholarship Award. This year, we were fortunate enough to have two very deserving candidates and those deserving candidates are MD Zia and Mr. Austin Lean. If you gentlemen would like to make your way to the stage, I'll go over a quick bio. MD Zia has a BS and a master's in plant pathology from Sheree Bangla, Agriculture University in Bangladesh where he is an assistant professor in the Department of Plant Pathology. Zia started his PhD program on plant pathology in Dr. Mohamed Khan's lab in 2018. He is currently working on identification and management of major sugar-beaked diseases such as sacroscal leaf control, rhizectonia, and crown rot as well as sclerotinia leaf blight. Since coming to NDSU, he has co-authored nine peer review publications and highly recognized journals. Austin Lean is a narrative of Fisher, Minnesota and a fourth generation sugar beet farmer. He obtained his BS in agronomy from the U of M in Crookston, his MS in agriculture from Washington State University and is currently working on his doctoral degree in plant pathology at the University of Minnesota since summer of 20. Austin is being advised by doctors at Shukchanda and Brent Arons. Austin's research is focused on developing a microplate-bast assay for assessing fungicide sensitivity and understanding molecular basis of fungicide-risk resistance in Sikospora. The outcomes from his research will be valuable in formulating effective fungicide spray programs for managing Sikospora leaf spot. Gentlemen, congratulations. All right, our next award is our Meritorious Service Award that is being awarded to Mr. Norm Cantna. Norm grew up on a dairy farm in Central Wisconsin and graduated with a BS degree in general agriculture and a minor in natural resource management from University of Wisconsin River Falls in 1982. Following graduation, Dr. Joe Giles invited Norm to join the NDSU soil science department and conduct research in sugar beet. Norm and Joe worked on a wide variety of topics including seedbed preparation, soil compaction, tillage methods to name a sampling of the projects. But you know Norm Bessver is organizing an operation of grower, planter, test stand clinics throughout the sugar beet growing regions including Minnesota, North Dakota, and Eastern Montana. In addition to working with Dr. Giles, Norm collaborated with doctors Laura Overstreet and Amitava Chatterjee. Norm intends to retire in April of 22. Norm, thank you for your decades of dedication and service to the sugar beet industry and congratulations on your upcoming retirement. It should sing a song or something. This has been a great ride. I came here almost 40 years ago and I didn't know what I was getting into. I can appreciate her working with Norm. It's been good getting to know people from the research side of it, the industry side of it, the seed side of it. I've learned and experienced something I'll cherish forever. I get to work with colleagues, I've got my brother, Al Dexter, Joe Giles, every single one. So it's been a great time. I'm gonna miss it, so thank you. And last but not least is my pleasure to announce the winner of this year's Distinguished Service Award to Mark Patel. Mark started his career in the sugar industry in October of 1990 as an agriculturist at Southern Minnesota Beet Sugar Cooperative. In 2008, Mark accepted the production agronomist position at SMBSC and was responsible for the official variety trials and various other agronomy projects. In December of 2015, Mark accepted the research director position for SMBSC and Spreckle Sugar. Mark is a member of the Sugar Beet Research and Education Board, American Society of Sugar Beet Technologists, Board of Directors, and the Minnesota Agriculture Fertilizer Research and Education Council. This past summer, the EPA approved Alter Blazer for Water Hemp Control and Sugar Beet based on Ms. Emma Bird and Dr. Tom Peters research. However, Mark coordinated an assessment of yield loss caused by Water Hemp across the three different co-ops that was absolutely vital in supporting the technical data and securing the emergency exemption. Mark's meticulous habit for detail is evident in other projects in his field research, which he has either directed or facilitated for over two decades. Mark, congratulations on behalf of the SBIRB on this award. I'd like to thank the Sugar Beet Research and Education Board for this award and recognition today. When I look at the list of past recipients of the Distinguished Service Award, I'm truly humbled and appreciative to receive this award. I'd like to thank my family, my wife, Beth, my son, Sam, and the daughter, Megan, for their support and encouragement over the years and allowing me to be part of this great industry. I need to thank SMBSC and Todd Dacellius for all the opportunities he's provided for me over the years. I want to thank our research team at SMBSC, David Metler, Cody Grun, Gary Lindahl, Bob Johnson, and Lindsay Nos. It's truly my privilege to work with each of you every day. Joe Hastings and Mike Medsker, thank you. You two gentlemen are the example of what has created both this industry. Dr. John Lam, thanks for all the collaboration, discussions, and encouragement over the years. Tom Peters, thank you for all your collaboration and encouragement over the past several years. I also want to thank so many members of this industry that are present here today or joining us online for the encouragement, cooperation, and collaboration with us over all these years. Thank you again.