 You know, as we look at this industry, it's a multi-billion dollar industry up and down the valley and it has direct economic impact on every community throughout this region. And so we're really proud to be able to partner with you and to be here as part of this program. We want this program to be relevant. And so if you've got topics and things that you'd like to see us conduct research on, conduct extension programming with, be sure to talk to Mark or Muhammad or Tom and let them know what your needs are. Communicate with them. We really want this to be relevant to you. I also want to give you a just a couple of quick personnel updates. And on many of you are kind of watching NDSU here. We will have a new president come July 1. And so the search committee for the president is working to identify candidates that will come into campus for interviews later this month and early in February. Dr. Brashani, 12 years of service at the institution, remarkable run that he's had. And we're very grateful for his leadership as part of this institution. And certainly from my standpoint for his appreciation for agriculture and in the impact that agriculture has on the state of North Dakota in this region. In addition, we are also beginning a search for interim director of program leader for Ag and Natural Resources with an extension. Dr. Charlie Stolt now took a job as director of the University of Nebraska's extension programs beginning January 1. And so we'll look forward to having a new director for our A&R programs and extension later this year. Mohammed, with that, I'd like to welcome everyone and thank you for the opportunity to be with you this morning. Thank you. Thank you, Greg. Mr. Eric Erdman, the chairman of the ARTI board. Thank you, Mohammed. Good morning. On behalf of the Sugarbeet Research and Education Board of Minnesota and North Dakota, I'd like to welcome you to our 52nd annual Sugarbeet Research Reporting Session. Thank you for taking this opportunity to attend, whether it is here in person or virtually and to our friends and colleagues attending online from around the world. In Austria, Germany, Serbia, Ukraine, New Zealand and of course in Canada and here in the US. The Sugarbeet industry is very vibrant and economically important in this region where over 55% of the Sugarbeets are produced and results over $5 billion of total economic activity. Our producers are strong proponents of research, are very progressive and rapid adapters of technology. We are thankful to our producers for their continued funding and to our researchers for consistently providing practical, sustainable and economical solutions to our problems. Thank you for participating today and I hope you have a productive and educational meeting. Thank you. So our first speaker will be all the way from Sweden. Dr. Desiree, she will discuss focus areas for sustainable intensification of the Sugarbeet crop in Sweden and Denmark. Dr. Desiree, I do not know how to pronounce her last name. Well, managing director for the Nordic Beach Research who will be Denmark. Good morning, everybody. My name is Desiree Böjstotto and I'm very honored to be invited to the 52nd annual Sugarbeet research reporting session. I'm the managing director, as Mr. Kanne said, of Nordic Beach Research since 2016 when I switched to Sugarbeet after a broad engagement in the crop production in this region. First, I will briefly introduce the Sugarbeet production in our part of the world. Then I continue to talk about sustainable intensification and one challenge we struggle with in that respect. Before I sum up with the conclusions, what to do to continue the development in our crop. So, first to decrease the distance today, I would like to put you on our map. See if I can do that, yeah. And found this nice overlay of North America to Europe. So here in the very center, you can see Fargo. And if we continue northwest, like 1,300 kilometers, you finally come to Denmark and Sweden. My everyday work is far from where you are growing Sugarbeets. We will see here today if we, despite the distance, might have something in common. Here is a close-up of, I'll see if I can, oh. Perfect. Here is a close-up of Denmark and Sweden. In the area, we have around 60,000 hectares, which is equivalent to 148,000 acres Sugarbeet, grown by 1,700 farmers that delivers to three factories. We also grow organic Sugarbeets to a small extent, and mainly to serve industrial customers with locally produced Sugar also for this market segment. So NBR, Nordic Beet Research, is a small institute based funded by the growers in Denmark and Sweden together with Sugar Industry in our two countries. About 40% of our funds come from disagreement and the rest from company trials, including variety trials, plus external research funds. We are today 10 colleagues, including two PhD students, and you will soon meet one from my team, Anilistet Hansen, who is today talk about weed control in our area. Well, we work mainly in applied research projects with close relation to the challenges Sugarbeet growers face here. I will come back to that, but firstly, I would like to show yield data from our two countries. Here you see the Danish and the Swedish yield development the last decades. And maybe most interesting might be to see the trend that we have an increase of about 200 kilograms Sugar per hectare in year, equivalent to 80 kilograms per acre and year. And we say that about half of this increase is related to the genetic material in the varieties. This is actual campaign data and the Sugarbeet crop is a good example of how genetic potential is well handled by the farmers. You also see that the levels differ between our two countries. And for that reason, today I will focus on Sweden. Now to sustainable intensification and wise people researchers have stated that food is the key to the global goals for the future. Our one definition of sustainable intensification is to increase yields without adverse environmental impact and without the cultivation of more land. It is not an easy task and in combination with the strategy for Europe, the European Green Deal, we are certainly challenged. To put it in our context, we see both challenges and possibilities. Looking back, the sugar yields, I said, increased with about 80 kilograms per acre in year. And this is by increase of root yield since sugar content is actually very stable in our area over the last 35 years. Here we might have genetic potential for the breeders to develop and to further develop. With more focus on sustainability, sugar content is interesting to increase the importance of in the price negotiations, I think. You get what you pay for. So we have a large engagement from the plant breeding companies in our region. And I said half of the yield development comes from the varieties. And they test every year 80 varieties in our two countries. And on the recommended list, we have about 25 varieties. The growing season is prolonged and looking 60 years back, we now have in average 11 days longer growing season in the area due to, well, the climatic change. The management of the crop is improved small steps at the time and we have identified areas of certain concern and included them in our NBR strategy over time for future development. And I have chosen fertility and plant nutrition establishment with an insect control you will hear more about from Annelis Petansson in a minute and control of leaf diseases, harvest and storage as certain focus areas. And last but absolutely not least, the growers that are devoted and engaged in the crop to invest and apply knowledge is a success factor. We have a lot to talk about as you see. However, I need to restrict myself to only talk about one of our challenges today. And well, I chose to start with the basics. And here is actually a typical sugar beet field outside my window at this time of year. A fertile soil with excellent status waiting for the sugar beet to be established in end of March, beginning of April. Or at the end of June, it looked like this. We saw this in large scale five years ago when many hectares of the very best areas in Sweden looked like this. We started case studies to follow up the soil status soil structure. We analyzed for nematodes, afenomyces and everything we could think about. The season of 2017 was wet and then it was followed by 2018 that was really dry and the problem occurred even one month earlier. We continued also the following year and looking back, it is almost embarrassing that we couldn't conclude that it all was about soil fertility from day one. In all fields being sampled, we found low pH and low calcium availability, sometimes in combination with afenomyces, we had low phosphorus and potassium status, less common to apply the fertilizers with row application so lower conductivity closer to the plants and not always was sodium applied in these fields. And for different reasons, we are facing this at significant more locations compared 20 years ago. So it's a little new problem. If I had more time, I could have showed you data and also had discussions with you to hear about your experience in this field but maybe next time. Well, I see one large reason to be cost savings supported by the environmental regulations to prevent further eutrophication of the Baltic Sea. Also the long-term financing of land is part of it with about 40% of the land rented in the sugar beet region nowadays and thereby less interest to invest long-term for the growers. However yield potential is strongly negatively affected and so is profitability of cultivation of sugar beets but also canola and in worst cases also in cereals when it looks like this. So under unfavorable conditions, the availability of nutrient is limiting growth together with less conductivity in the soil favors this, the effinimizes and this gives less growth rate to the sugar beets and it all sends in low yield potential. Soil fertility was studied since 1957 that is for 64 years in four long-term fertility trials with sugar beets in the rotation every fourth year at the university and we've, well, eventually we've found out about it and the trial and these trials and ask for data. And as you know, phosphorus is a complex subject and the plant needs in comparison rather small amounts at the same time as the soil often holds sufficient amounts. It is all about availability which varies a lot between soil types but also depends on water supply. In the plant, the phosphorus is as necessary as nitrogen to reach high yield levels and typically the sugar beet takes away 20 to 30 kilograms of phosphorus and the soil in the road soon, so contains often many 1,000 kilos. However, the system is not easy to affect and the movement extremely slow. The results of these trials shows if we set removal of phosphorus potassium as a standard so 64 years without applying any phosphorus and potassium in the rotation we have actually an impact on the sugar yield with 36% lower yield and if we apply removal plus 15 kilograms of phosphorus and 40 kilograms of potassium and now I have to say this is per hectare so divided by two and a half to get it on acres for you, we have an impact of seven and if we apply the double plus removal we have a positive result of 20% higher yield. So this well pays for the costs of the applications. So to conclude and emphasize the most important limiting factors we see in soil fertility to reach the potential yields all in all fields and in all parts of the fields is row application of a full nitrogen, phosphorus and potassium plus micronutrients fertilizer at sewing. And here you see, well, it's, sorry, it's hidden behind here. You see that here is the row and you can see the sugar beads seed here and here we have the row application of fertilizer. So this is our recommendation to put it, well, we sew at two centimeters and then six centimeters below the seed and six centimeters beside the row, we apply the nutrients. Maybe it's the same way you do it. We also aim for a high availability of phosphorus and we, this mean to have a positive balance over the crop rotation and this is in all parts of the field. So one method is to use variable application rates both for phosphorus and potassium and precision farming and variable rate applications for lime and also phosphorus and potassium to lower the field, the infield variation is used on about 20% of the sugar bead fields. And here we have room for improvements. But here is a typical example from 2009 and we have applied, this is for phosphorus and it was applied over 10 years and you can see that the maps are improving. Well, it's important when you grow sugar beads to have pH at seven, 7.5 and then when you have reached that level continue to apply lime once in the rotation. That is our recommendation. We need to keep balance of potassium but also apply sodium, sorry. And that can, we can see in some field that the higher conductivity, we can have less affinimizes. But this is an area where we do some projects and some research around. Yeah, well, we did not talk about infiltration and drainage which is also very important for the soil fertility. Most fields are drained in our region and in relation to land prices, it is the highest priority to keep them in best standards. And here is how it can look like in our countries. But to be able to reach the goals of the future, feeding the world, we all need to manage the land resources at our best and use the best practice. And in this context, soil fertility and sustainable intensifications plays the major role. So I say thank you very much. Our next speaker will also be from Denmark, Dr. Annie Lisbeth Hansen. She will speak on managing weeds in sugar beads in Denmark, Sweden and EU, successes and challenges. Good morning, ladies and gentlemen. I will give an introduction to managing weeds in sugar beads in Denmark and Sweden with an outlook to European Union and look into our successes and challenges. At our European latitudes control of weed is one of the most important factors to ensure a high sugar yield. That is because weeds compete strongly with light, nutrients and water with the crop. Danish tribes show already at 2% weed cover in June and risk of 3% yellow loss as seen in the figure. Besides the direct loss of sugar yield, poor weed control will hamper the harvest, increase the tear, increase infestation of pests and diseases, and moreover, it will increase the seed bank in the soil and escalate weed problems throughout the whole crop rotation. To a large extent, our weed control is determined or regulated politically. Some points here to mention are, back in the 18th and 19th, much research and trial work were performed with a successful development of low-dose strategies where optimal timing is combined with the lowest possible dose of herbicides while maintaining a high efficacy. Especially in Denmark and Sweden, there has been extra focus on reducing the pesticide use and the amount of pesticide used has been halved from the 18th to year 2000. Approval of prime protection products is regulated in EU. The approval is very strict compared to, for example, the American system, which means that we have much less available pesticides for outdoor use. The given approvals are valid for a limited period and then they must go into a renewal process where new criteria might have been put up. Following the EU approval, the substance can be undertaken national approval regarding focus areas. In Denmark, occurrence of metabolites in the ground border is an issue and in Sweden, the surface water is taking more into considerations. And since 2014, all professional users must implement integrated pest management and the eight principles. For example, look into crop rotation, monitoring and decision support systems. To increase a sustainable use of pesticides in agriculture in EU, EU has set up a set of goals for the next 10 years, which among others is to reduce by 50% the use and risk of chemical pesticides by 2030 and to compare a baseline from that on average of the years 15, 16 and 17. Also to mention here is that the European Union will boost environmentally friendly practices among others by aiming for 25% of total farmland on the organic farming by 2030. In our region, most conventional weed control strategies consist of one pre-emergence application as an option and three to four post-emergence treatments beginning at the first flush of the weeds at cartilagin stage. The cornerstone active ingredients are fenmeida, fenmeida, itufumacet and mesumetrone. And depending on the weed species, for example also trifosulphurone and clomosulphurone and clomosulphurone is added. Some sugar beet growers will perform a mechanical cleaning after the end of the herbicide program before grow closure or perhaps even before the last spray. In some countries, the conventional smart system is available where an ALS total and variety is grown and two applications of the ALS herbicide commission one plus a conventional herbicide is applied. And more about the active substances that we are using for the control of dichot weeds. Here we see an overview of the estimated percentage share of treated area with active substances for Sweden, Denmark, Germany, France and Netherlands. Where we see that often the most often used herbicides in the dark green and in the lighter green colors the herbicides to be added. However, the substances in the lighter colors must not be considered to be of less importance as they control specific weed species which is also not controllable. In the bottom of the table, the estimated use of mechanical weed is noted and probably Denmark, Sweden and France are in the front using most hoeing in these years. From this list, we have lately lost substances for some dismedi fame which was not reauthorized. Here in yellow, you see the ingredients which are either under renewal or which are soon entering the renewal process. And as you can see, there's quite a lot. We support the work in our RRB weed control group in order to support the renewal process. For example, are we at the moment supporting the renewal process of trifle-stilforone resistance? Another important challenge is the increased development of herbicide resistant weed species. The graph shows the increase of resistant resistance to five herbicides mode of actions. Different herbicide mode of actions have different tendencies to select for resistance and this we must be aware of and seek to prevent. ALS inhibitor herbicides group two are the most prone to develop resistance and dominates among our weed species, especially in the grasses. And in the same time, ALS inhibitor herbicides are frequently used in the whole crop rotation. It is a challenge to avoid building up resistance in weed and demands a good management. Moreover, the likelihood of having less available active ingredients in the future make prevention even more difficult. In NBR, we control tricep each year. As an example here, we are studying what we are studying, a show an average of two dangerous trials from last year. The orange bars show percentage efficacy. The basic treatment is seen in the table below for revocations, which consist of in total six litre bitanil containing filamentifam half a litre nortone containing itufumacide three litre galatex containing metamethone 10 grams safari containing trifosulphurone and half a litre of oil per treatment. In the control treatment there was an average 46% weed cover observed in June and the dominating species were fat hen, foots, parsley and cleavers. In the control, yeah, we can see that we lose. Here in the red ring, we can see that we lose efficacy when we are leaving out death medifam as in entry from entry two to entry three. And the problem is that death medifam was banned at the renewal process and we have left, we have, death medifam was banned, but we have left to use fin medifam. Adding clomazone helps, however, on the efficacy. We lose, if we risk to lose fin medifam, which is also at the renewal process, but it is foreseen that there would be no problem, but if we lose fin medifam, we will have quite a lower efficacy as we see here. Adding clomazone and morosafari and also metagon will help on the efficacy. As an overall, we can see that we are in the field where we are dealing with a delicate balance and timing is very important. Studying the combination of mechanical and chemical weed control, here I want to show results from a sweetie set of trials where it was studied how much herbicides we can save when we perform three row cleanings. A 12 row machine with GPSRTK guidance on both the tractor and the row cleaner was used. Distance to the bead plants were adjusted as seen on the small pictures below. Where the distances started are 4, 8 and 12 centimeters. Healing was studied with the aim of cover the weed so it was moved into the row and the bead plants were covered up to 70%. In this study, number of weed accounted in the row and not between the rows because between the rows the weeds are controlled in an efficient way. But the question is how we fight the weed in the row. In the figure to the right, one dose of herbicide is applied at first weed flush T1 followed by three mechanical row cleanings. We have 10 to 20 weeds per square meter in the row. This is not good enough. But we can also see that the picture is getting better the closer to the row we get. And even better if we move some soil into the row by healing. In the middle picture, we have applied two half doses of herbicides and it is getting better with less weed left. With less weed left. However, it is not quite good enough yet. In the figure to the right, combining two herbicide treatments at T1 and T2 with mechanical cleaning, we are down to an acceptable level of weeds left. Healing gives 30 to 40% less weed in the row. Other trials have shown similar results. Mechanical weeding is more successful when small weeds up to four leaves are treated twice and it's treated twice or more and in dry, under dry conditions. Measuring sugar yield in such trials show that the closer, the later and the more often mechanical cleaning are performed, the more sugar yield is lost due to the treatment. But it is around one to 3%. And in the presence of weed, the weeds are more expensive. Coming to the future weed control. Will to a higher extent be based on integrated solutions and many promising developments are ongoing. Overall, on our latitude it is likely that we will continue to see a reduction in the approved active substances and also in the allowed doses. Machinery for mechanical weeding exists today with more robust constructions which perform with higher precision steered by cameras or GPS RTK. Their conventional smart system developed by KWS and Bayer varieties now also sold by Cisfenhaeve beta seed and DLF bead seed is a system available in some European countries. Commission one was launched in 2016 in Sweden, in Lithuania and in Finland in 2017 with a dose of one liter in a single or split treatment. There is an advantage using the system where the weed pressures were high or whereas many weed beads. The system is used in some countries. For example, Lithuania, where it is used in about 50% or above 50% of the area and in and above 30% of the area in for example, Finland and Spain and in Sweden approximately used at 10% of the area. Coming to precision farming, we go from broadcast to smart spraying using camera, GPS guidance, machine learning algorithm and weed recognition provide new opportunities for reducing the use of pesticides. In Denmark, we are working with new spray equipment and band spraying. Also, we are working with for example, spot spraying. So spraying just around the bead plant as seen on the photo to the right on the UV light. The robots are coming or they are already here. We are working with two Danish types, aqua-intelligual body, which is an autonomously robot control with a computer which can carry various equipment with a three point hitch. Also the farm dried, a fully automatic robot driven by a solar panel, performing sowing with GPS positioning of the seeds and which perform weed cleaning. Also there exist other types of robots in Europe. Yeah. With that, I will end my presentation and thank you very much for your attention. Annie, when do you estimate there will be widespread use of robots in Europe for weed control? There's already some robots have been sold and I cannot say when it will be used in a large scale, but it is in a quicky development and there are already a hundred or more sold and it's running and there are quite a good satisfactory experience with that. Over these. Other questions, can you please put it in the chat box? So it seems like the future in Europe, there will be reducing pesticide by 50%. Some of it will be organic. You'll probably have to use robot to manage weed successfully for sustainable sugar beet production. Mark, you want to get ready for the next speaker please? This will be Dr. Debbie Sparks from the University of Nottingham. She will discuss the impact of canopy architecture on radiation use efficiency of sugar beet. Thank you, Mohamed, and thank you very much for the invitation to speak at your meeting today, much appreciated. So in my presentation, I'm going to discuss ongoing work on canopy architecture and sugar beet. And I should start by acknowledging from the beginning that Lucy Tillier, the PhD student has done all the hard work on this project. And when I say we, I usually mean Lucy. So I should say that from the start. So there's no misconceptions. So the project really started with thinking about the different canopy architectures that we've seen coming through in new varieties in recent years. The relationship between intercepted radiation and sugar yield is long established in the absence of other limitations. And while there's now some discussion that we sink limitation in sugar beet, at least in the UK, we still find that this direct linear relationship holds true. So we were really interested in these different prostrates and more upright canopy architectures that have we've seen in recent years. I can just try to get a pointer. So you can see in this photograph here, we've got one variety that's much more upright and one that's much more prostrate. And we set out to ask, how does canopy architecture impact lighted deception by the crop, radiation use efficiency, and of course yield? As I said, this work is part of a PhD project. And some of you may have already met Lucy Tillier at an event such as the IRB Congress. So she's doing the hard work, as I said. In terms of monitoring our canopy response over time and canopy closure and lighted deception, we're unable to use drones at our site in Nottinghamshire because we're directly under the flight path at East Midlands Airport. So we had to be creative and mount some sensors on the tractor here. So we've got a downward-facing camera and also a downward-facing sensor monitoring things such as NDVI and NDRE. And at the bottom of the screen, you can see a photograph. So once we take the photograph, we then use image analysis software. And here the green of the canopy has been marked as red and then we count the pixels to work out percentage canopy cover. And then that data is plotted on the chart on the right. And you can see here, we're looking at four different or three different canopy architectures. So a prostrate, a very flat canopy, a more upright canopy and two varieties that had a more intermediate canopy. And in this first year, this is 2019, you can see that during the period from about 70 to 120 days after serving, the varieties were quite different in terms of their canopy cover. And therefore their light interception and converting that light into sugar with the two intermediate varieties intercepting more radiation than the prostrate and the upright varieties. Of course, we wanted to understand the reasons behind that. And one of the measurements that Lucy developed was working out the canopy angle or the PTL angle. So by carefully taking photographs in the field and working out the angle between the most upright and the insertion of the PTLs, we calculated the canopy angle. And what you can see here is that the prostrate, because there's a larger canopy angle from the upright to the insertion of the leaf has the highest angle. And then we've got the upright at the bottom and the intermediate in the two. So clearly there are big differences in the architecture of the canopy. And this is likely to explain some of the differences in canopy cover and therefore light interception prior to canopy closure. But we were more interested in radiation use efficiency and therefore yield than canopy cover in its own right. So in order to calculate radiation use efficiency, we did that the old fashioned way really by sequential harvests over time to measure biomass and then calculate radiation use efficiency. So you can see the photograph of some of this hard work in the field and here on the right, I plotted total biomass over time for the four different varieties. And again, we can see that as time goes on and as we get towards the end of the season, the two intermediate varieties sort of pull away in terms of their total biomass in tons per hectare. We then took this data along with the canopy cover data and the local Met Office data for light incident radiation. And we use that to calculate radiation use efficiency. And that's shown on the chart on the left. And we can see that the, so what we've got here color coded the intermediate varieties prostrate and upright with the dots color coded on the chart. And the figure you've got here 1.82 that tells you that for every one gram or one megajoule of light intercepted is producing 1.82 grams of biomass. So 1.82 grams of biomass per megajoule of radiation intercepted. And what the stats showed us was that the two intermediate varieties had a higher radiation use efficiency than the prostrate and the upright. They were similar to each other but lower than the two intermediate ones. And that came through in yield very clearly. So you can see again that the two intermediate varieties had a higher yield, much higher sugar yield than both the upright and the prostrate. And surprisingly to us, there was no real difference between the upright and the prostrate architectures. So we clearly were picking up some differences but were these differences due to canopy architecture or to some other genetic differences between the varieties that we had selected? You know, we've only got four varieties here. One example of a prostrate, one example of an upright and two intermediate. So we couldn't be sure that it was about canopy architecture. There could have been some other differences. So moving on into the second year of the study. Unfortunately, this coincided with the beginning of the COVID-19 pandemic which meant there were no field experiments for us in 2020. But later that year as the restrictions eased a bit, we were able to conduct a controlled environment study which looked at the photosynthetic capacity of the different varieties. This time, because of space constraints, we only used one of the intermediate varieties but we had the prostrate and the upright varieties in there. And what we've got on the chart is the three varieties and then leaf four and leaf seven. So leaf seven is the youngest leaf. So leaf four emerged before leaf seven. And actually statistically, there was no interaction between a variety and leaf number. So I've just included the chart for completeness. And then the tables on the right here show us that where we've actually got significant differences. So this A max value is the maximum photosynthetic rate that we measured of these different leaves and different varieties. And you can clearly see here that the youngest leaf had a higher photosynthetic capacity than the older leaf, which is as we would expect. And we can also see on the table above that the intermediate variety had a higher photosynthetic capacity or photosynthetic rate than the prostrate and the upright varieties. And although there's a numeric difference between those two, it wasn't a significant difference. We measure lots of aspects of photosynthesis and just want to present two more. One is the light saturation point. So this is the point at which there was no further response to light and the canopy was saturated by light. And again, I've done the same thing and showed you the chart for completeness and then the tables for the stats purposes. So again, we can see that the younger leaf has a higher light saturation point than the older leaf and the intermediate variety had a higher light saturation point than the prostrate and the upright. So what that means is in the field, we would expect the intermediate variety to keep photosynthesizing at higher light levels. I should just say that the absolute levels that you're seeing here are not that high and that's because the CE or the control environment room cannot produce very, very high light levels. But in terms of the acclimation of these varieties, we're seeing the relative differences between the different canopy architectures. And then the final thing with regard to the photosynthetic capacity is something called the quantum yield. And this is a measure of photosynthetic efficiency. So it's a dimensionless characteristic but it's taken to reflect photosynthetic efficiency. There were no differences between the leaf age at this measurement. But again, you can see the intermediate variety has having a much higher photosynthetic efficiency than the prostrate and the upright. So the control environment data seems to fit very well with the field data in that the intermediate variety has a higher AMAX, a higher quantum yield and higher light saturation point. But we still don't know if this relates to canopy architecture. So this year we set out to sort of test this more in the field. The field study was expanded to include more varieties. So we had I think six varieties in total. And then also some direct sort of manipulations of the canopy. We did this in two ways. The first way on the left-hand side, I don't know how well you can see this photograph but we've actually planted alternate rows, one of a more prostrate and one of a more upright variety in order to create a sort of mixed canopy that we hoped would be more efficient than growing either the prostrate or the upright on their own. And then on the right-hand side, we took the same genetic background. So one variety, one of the intermediate varieties and we artificially manipulated the canopy angle. And that was done very meticulously by Lucy who went out and used some sort of pegs, like tent pegs to hold down the leaves so that to create a more prostrate canopy and also use what we call these sort of cages to push the canopy more upright. And then over the year, Lucy's measured lots of very detailed assessments of light interception, light capture and photosynthesis on these varieties and these treatments. In addition to measuring things like canopy cover, NDVI and NDRE from the tractor as we did two years ago, we also added an extra sensor so that we could measure reflectance from the canopy because you hadn't really considered that before. We'd sort of assumed that all the canopy architectures would reflect the same amount of light, but we're now looking in detail at whether some canopies absorb more than others and some reflect more than others. So that's being studied as we speak. And then Lucy also used what's called the ASD field spec to take a wide range of spectral indices over the season. So we combined measurements at the canopy level with measurements at the leaf level. What I'll say at this stage is that the results look really interesting and consistent with our previous findings, but Lucy will be presenting these in detail at the IRB Congress in June. So I don't want to steal her thunder here and I would just say watch this space. I would just like to finish with this image created by Annie who's another member of our beat team at the University of Nottingham. She made this for the Christmas card after doing some very cold measurements in December this year just before harvest, although not as cold as North Dakota of course, but pretty chilly for us here in the UK. And with that, I'll finish my talk and I'm very happy to answer any questions. Our next speaker will be Dr. Bram Hansen from the IRS in Netherlands. And he will share with us the Stemculium Beticula because of yellow leaf spotting sugar beet. And this is more or less to make our producers in the US aware of a potential disease. It has been found in the US on spinach and table beets. So all our producers look at this. If you see any symptoms like this reported to your agriculturist and then we at the university and research centers will become aware of this as well. I'm very pleased to present something about Stemculium Beticula which is kind of, well, a new disease in the Netherlands which almost started his career at the same time as if I did start my career as a fighter pathologist in sugar beet. So we grow up together. A picture of the Netherlands we're quite close to Germany and the UK. A large part of our acreage is below the sea level and we grow 48,000 hectares. So about 200,000 acres of sugar beets. That's more than tulips we grow. We still have those nice pictures of tulips and windmills but I prefer the sugar beet and a windmill. Our sugar yield is about 30.7 tons a hectare this year. We have two factories, one in the south but also our institute is located, one in the north where we process all the sugar beets in about 120 days at each factory. For the 40 diseases in the Netherlands we grow sugar beets for about 80 years and we have a long history with 40 diseases especially powdery mildew and the rust and peramelaria which are quite mild under our conditions. In the 1980s of last century we get some experiments. The Cospra moved from Italy, Greece via France and Germany to the Netherlands and it caused up to 40% of sugar yield reduction. And that's quite a lot. So the main focus was in the past on Cospra management. In 2007, we found some fields which were applied with fungicides, mostly product allegro and opistine which contain a particular soil and it turned already in September quite brown and yellow. And we visited those fields because the farmers were complaining they could not get manned their Cospra infestation or their Ramallaria infestation. When we entered those fields, the first plant was quite yellowish on those leaves which is not typical for Cospra or Ramallaria. We examined the spots on those leaves got to the conclusion that it's nor Cospra, nor Ramallaria, it's some other disease but we did not know which one. So this first fields which were quite located in the northeast where the first we have large infestations and yield loss due to this fungus. Those spots, we call them initially yellow spot disease, are quite typically irregular and yellow and when you have some back lights from the leaves they are quite shiny, so it could also be a virus. So we checked for viruses, we checked for nuisance deficiency, we checked for bacteria and we checked for fungus and we isolated quite a lot of fungi from those disease leaves. With the tiny yellow spots, necrotize from inside out, grow in size and when you have a lot of spots they come together and you cannot be turned brown and it's not very efficient in picking up the sunlight and transform it into sugar. If I repeat, those spots are irregular. So the size, when it's round, it's not senphillium, you can say it by heart. It's irregular in size and in the middle or somewhere in the middle of the yellow spot you have a necrotizing tissue. It turns later on a brownish spot which can be nicely rounded but also quite irregular in shape and often you see those yellow spots and dots next to those brown necrotized tissue. Very typically irregular in size, the yellow spot and then the brown dot in the middle, the necrotizing tissue. So it's the growing fungal tissue in the leaf tissue and you have some interaction with some toxins from the senphillium which is causing the yellowing in the leaves. Very nice symptoms for a phytopathologist, for a farmer when it goes quite quick from infection to spots, it's about one week. So you can have multiple generations of those fungal infestation in your field in the season and that's causing without any management the total collapse of your canopy. And later in the season, so about late August, early September, you can see both the yellow spots on the brown tissue. Initially it started with a few yellow spots on those leaves, hard to find and it's easy to mix up. It looks like insect damage when you have those nice round yellow appearance or the other side of the leaf where you turn the leaf upside down, you see that some insect has eaten away some of the leaf tissue, causing a yellow gland on the other side of the leaf. Also some insect stings with puncture. The leaves suck out some tissue. The tissue could be to reacts with a yellow surrounding of those things. The older spots look quite like Pseudomonas. Pseudomonas, the bacterial leaf spot, also causes those brown spots which has quite a weak brown tissue and you have those breakage of the leaves quite easily. That's also with the brown spots of Stemphilium. Burned-in spray liquid. You often see it with herbicide folium mixes. When they are applied full sunlight, you have some droplets which burn in to the leaf tissue and leaving some scars. It also quite looks like those Stemphilium yellow spots but are not so. There can be some misdiagnosis for the Stemphilium. Also manganese deficiency. When you turn the leaves up to the sunlight, you have those yellow spots, the yellow clouds in the leaves. Even when it's a quite old manganese deficiency, you can have also brown dots inside those scars which can be misunderstood for Stemphilium. So be aware when you're scouting in your field. Not all yellow spots are Stemphilium spots. We isolated Stemphilium from those yellow leaf spots and when we grow them on nutrient media in the lab, we saw those nice Stemphilium spores and well, try to identify those Stemphilium species on that. And that took multiple years before we got to the conclusion, it's a new species. We have to give it a name. So we named it after Shukerbeef. But also this identification, we did it together with the Vestodeig Institute in Utrecht, the Netherlands. This identification caused a complete revision of the genus Stemphilium. But in the collection of these Vestodeig Institute, also some old strains of Stemphilium which were not identified before, but collected in the 70s and 80s of last century originate from Canada and the US. So, Stemphilium beta-Cola is already present over there since a long time. And when it wants to acquire enough when you have enough spores on a hectare, you will see the spots anyway. In our field trials, it can cause up to 40% lower Shukerbeef. So under favorable conditions in the Netherlands, it is as aggressive as Cicospora beta-Cola. So that means we have a lot of effort to reduce the impact of this fungus on our Shukerbeefs. The first thing we were asked to investigate was, is there a relation with those varieties where we find a lot of Stemphilium net? Is it variety dependent? So we can skip out those most susceptible varieties and then go on like we're used to. So I had some isolates from all over the country and you see that there is a difference in ranking per isolate. So isolated one and isolated two are quite close together. Isolated in the northeast of, in the northwest of the country and isolated three was isolated in the northeast of the country. And you see we have on top, so least susceptible isolate one and isolate two, variety rhino, which was blamed in the northeast would be most susceptible. And also for the variety isobelacate WS, which is the variety where we isolated three from. There it's most aggressive and on isolated one, isobelacate WS is, well, really unsusceptible. Also for the rhino, you see some differences. So that means when we have a small country, but when we have this distinct locations, un-distinct isolates, we get distinct ranking of the varieties. So that's quite problematic when you would like to give one advice to growers. So we treated Stemphilium like our Sacospora, we did a lot of research for fungicide efficacy and we put it in our monitor-based system. So we start monitoring for folio diseases after canopy closure, at least once a week. That's our advice. But well, the best practice is to do it once a week. Some farmers skip it and do it once a month. And then, well, they are most prone to make mistakes in the management of folio diseases. When we find some spots, tranquil spots, either Sacospora, either rust or Stemphilium, we have a fungicide application. And then we start monitoring two or three weeks after the application of the fungicides. And then we have a new application when we see an increase of infestation or a new folio disease. That means that we have one system for all folio diseases. And then we need to know what the product you should use for the management of Stemphilium-Bethicola. We have some products like Opostim, which is a product based on epoxy-conazole, which is quite effective against the Crosper-Bethicola, but it's ineffective against Stemphilium-Bethicola. The same for the Allegro, it's Chrysoxymethyl and epoxy-conazole. It's, I use it in my filtres as a control to have, to have no influence of Sacospora in those spots, but for Stemphilium, it's like you apply water. Then we have the Cipro-conazole and the Dipro-conazole. It's one plus, sometimes you have M-propidin next to the Dipro-conazole. You can have the second plus. And then we have the three pluses for the Bascali. So the SDHI, fungicides, the same for the propols, the free opyrum. So those SDHI's are much more effective than the triazoles and the strobrorins. Are we unique? No. Since 2007, the first years we thought we were unique having this disease, but since that time, I presented a few times at the IRB or other concessions on this fungal disease in Sukubi. And then I received some photos or even leaves from other countries, showing the typical symptoms of Stemphilium-Bethicola also in other countries. So the photos, the acknowledgement is in the photo itself. So we see almost the whole North-West, northern part of Europe, we can find the Stemphilium-Bethicola. And at the end, this is our goal, keeping healthy leaves up to the harvest. Are there any questions? One question for you, Bram. Do you think the pathogen will survive on the U.S. conditions? Well, the pathogen won't survive when it's dry. It needs a lot of humidity to infect the Sukubi leaves up to eight hours of lead. So we see the years where it's our typical Dutch summons. So we have a lot of rainfall, lower temperatures, so around 25 degrees. We see a lot of Stemphilium when we have high temperatures, so about 35 degrees Celsius. Quick drying up of the canopy, it's favorite Sikospora-Bethicola. So in years we have a lot of Sikospora-Bethicola, so our warm and dry and hot summons. The years where we have Stemphilium-Bethicola are wet and cool. For the winter, it can survive on the host issue. There are more hosts, like potato with also some green manure crops. And we are able to freeze it, the spores up to minus 80. So we can store those isolated at minus 80. So when winter is coming, quite easy, quite slowly, so there's not a quick drop of temperature, it can even survive those hot winters. And just for your sake, yes, it does survive in its present in the US, in Washington state. It was found in Table Beats and Spinach, and in New York, they have found it on Table Beats and it's very big on onions. All right, our next speaker will be Dr. Raj Majumdar. He will speak on cell wall degrading enzymes, originating from Rhizoctonia Solonii, increased sugar beetroot damage in the presence of Leuconostoc mesenteroides. He is with a group from Kimberley that include Dr. Strasbaugh, Dr. Goli, Wesky, Rakesh, and Rogers, Dr. Raj Kumar. Thank you for the invitation and good morning everybody. It's a pleasure to be here. So as you see the title of my talk today is Cell Wall Degrading Enzymes, originating from Rhizoctonia Solonii, increased sugar beet damage in the presence of Leuconostoc mesenteroides. As we all know that Rhizoctonia is one of the major causes of yield and sugar loss and sugar beets and sometimes an enter field can be wiped away by this pathogen, depending upon the nature of the infection. Whereas Leuconostoc, it's a free living gram positive soil bacterium, which has lesser effect on the root rot when it is present alone. But I think the devastation comes when Rhizoctonia and Leuconostoc, they are present together in the soil. That causes the most damage in it. You can clearly see here on the screen. So the most important thing now is that the genetic resistance against our solonii is highly limited and it will require identification of novel targets or mitigation strategies in the future. So as we know that Leuconostoc mesenteroides did not produce plant cell wall degrading enzymes. So we hypothesize that this increased damage caused by the close association between these two pathogens is caused by the plant cell wall degrading enzymes that is coming from our solonii. So the experimental approach was pretty straightforward. We had in-planter sugar beet root inoculation. So what we do is that we take a cork borer, we add a specific amount of the fungal or bacterium inoculum depending upon the treatments. And then we are also using pure exogenous plant cell wall degrading enzymes. So we collected samples, root samples, two different time points, early time points where one day post inoculation, two day and three day. And then these time points were used to for RNA-seq and metabolites. And then the late time point we use for disease evaluation. So the first objective was to evaluate the disease and how different exogenous enzymes that we had applied, how does it cause the extent of the disease in sugar beet roots? And then objective two was the mRNA sequencing where we are looking at the global gene expression in all three organisms at the early stages of infection. So the goal was to identify the early factors that actually caused this disease symptoms. And we were also interested in looking at the cell wall metabolites because that is one of our major targets. So how does it affect the cell wall degradation? And all of this thing led us to the identification of potential targets for future mitigation strategies. So this slide here shows you the effect of different exogenous enzymes. So the first one is control. And you can clearly see that when rhizoctonia is applied separately versus when leuconostoc is applied separately, the infection is quite higher with rhizoctonia alone. But when you put them together, you can clearly see that the damage is quite more. And then the lower panel here, you see three different enzymes that we had used. This is one of them is the pectinolias and polygalactouronase and the hycozyme. So hycozyme is a combination of all different kinds of plant cell wall degrading enzymes. And you can clearly see that PNL and PG, they had the most impact on the cell wall degradation and more rot in the roots. Suggesting that possibly these are the two enzymes contributed by rhizoctonia solanai is causing the damage when it is in close association with leuconostocs. And then we were also interested in looking at the gene expression of the rhizoctonia solanai at early stages of pathogenesis and you can clearly see. So each of these gene family that I'm showing here, pectinolias and polygalactouronase, these are big gene families. They have almost like eight to nine members. And we have been able to identify that which parallogs are the critical players that actually contribute to this increased rot. And these are actually boxed in red here. You can see that you can see quite a high expression starting from day one and then the expression increases and then the highest expression that we found was with polygalactouronase especially with this parallel that had the highest expression. We also see expression of the cellulose genes but I think the PG and PNL genes where they had the highest expression in terms of potential factors that cause root rot. But you can see at the day three that more numbers of cellulose genes are being turned on. So now we were very interested to see that what are the changes in global gene expression? And so what we did is that we did a combined RNA-seq approach and the whole goal was to identify how the interaction takes place between the host sugar beet, Raju Thunai Solanai and Leuconostoc mesenteroiris. So we had done for all three time points, one day, two day and three day post-conoculation but I'm just showing you some examples about the trend of the gene expression. And you can clearly see that the carbon and nitrogen metabolism related genes are highly upregulated in terms of the pathway enrichment besides the plant hormone signal transaction. So here I'm presenting a few examples of differentially expressed genes. The list is super big but I'm just focusing on a few things and what we can clearly see the very first and the top candidate that is showing up in our RNA-seq analysis is the polygalacturonase inhibitor and this is exactly what we had seen high expression of the Raju Thunai Solanai polygalacturonase genes. So it clearly tells us that polygalacturonase originating from our solanai is one of the major targets and that's why the plant is actually producing a polygalacturonase inhibitor protein to combat the situation. And this chart also shows you actually how the plant responds to a fungal pathogen versus a bacterial pathogen. So as an example, you can clearly see here that's proxidase 27 and all of this oxygen binding proteins. You can clearly see that it is highly upregulated when the plant is experiencing Raju Thunai Solanai not much with Leukonostar, basically it's little lower. So there is a differential response from the plant's perspective where the plant responds depending upon the nature of the pathogen whether it's a fungal pathogen or whether it's a bacterial pathogen. Similarly, we also looked at the global gene expression in our solanai at all different time points but I'm just giving here an example at one day post inoculation. And as expected that ribosome-related genes were highly upregulated because at the early stages of infection, the pathogen is multiplying. So it is quite expected that ribosome will go up and besides ribosome, carbon metabolism also plays a major role at early infection stages and we can see a lot of genes are being represented from citrate cycle and bioxylate and dicarboxylate metabolism-related pathways. A closer look at the differentially expressed genes is presented here in this table since we have a long list but I'm just showing you some example. And the one that is boxed here at the very top shows ADP and ADP carrier protein. This is a very interesting and exciting candidate. So when it first showed up in our data, I was wondering what is this candidate? What is known about this gene in other pathogens? So what we found is that, so a little background about this gene. So what this gene or this protein does is it actually recycles ADP from mitochondria to cytoplasm and during the early infection stages, it's highly dependent upon the source of energy and there are a few papers and one of the papers, it shows that in botanitis, when you actually mutate this gene, it loses pathogenicity. So this actually gives us a very clear idea what could be potential targets in the future if we have to take the RNA-based route. And besides this gene, bunch of other genes associated with ribosomal protein, which is quite expected and elongation factor. So this gives us some idea about what we could target in the future. Similarly, we also looked at the Lykonostoc genes. And there's a distinct difference between prokaryotic versus eukaryotic pathogens. And you can clearly see that in Lykonostoc messengerase, the pathway enrichment was for ribosome, biosynthetic pathways and selenocompound metabolism. So ribosome is quite expected because at that early stages, the bacteria is just multiply. Making more copies is the most important thing at the time and that makes sense that ribosome related genes are showing up there. But another interesting aspect which I find is selenocompound metabolism. So I was doing some literature such to see that what is this all about and why these are important. So selenium is a micronutrient which is present in both host and pathogens. But what happens is that selenium is very important for bacterial pathogenesis. And what selenium does is that selenium gets incorporated into cysteine. And selenocysteine, there's tons of literature out there in bacterial pathogenesis and the role of selenocompound or selenocysteines and how they affect bacterial pathogenesis both in mammalian system as well as in plants. So that's quite interesting and that shows that the difference between fungal and bacterial pathogenesis and what pathways are turned on. A closer look at the candidate genes that were differentially expressed. So we had the same way we had like one DPI, two DPI and three DPI. I'm just showing you a few examples of how the candidate genes look like in Lycunostoc mesenteroides. And the first candidate ATP synthase and translation initiation factor IF1. Lot of things are out also in the literature where people have targeted this gene specifically in bacteria to inhibit bacterial pathogenesis. So this gives a clear idea that what we can target in the future through potentially RNA-based approach, then as we see lot of gene expression-related changes in carbohydrate-related metabolism, we were also interested to see that what are the cell while degraded products showing up in our material. So as you can clearly see that these are the four different kinds of sugars that we have detected in our samples. Sucrose goes down with infection and you have the most reduction where you have both Rajatonia and Lycunostocs together. That makes sense because sucrose is the stored carbohydrate in the vacuose of the cells in sugar beet. And as you have more pathogenesis, there is a high demand of carbon and that's why the sucrose goes down. But interestingly glucose plus galactose and fructose, they go up. Why? Because these are the degradation products of the complex cell while sugars and thereby contributing to the production of more simpler sugars. And that's why we see increase in simple cell while sugars, which actually corroborates well with what we had seen with increased expression of the plant cell while degrading genes. Next, we were also interested in looking at the total carbon and nitrogen because what we see in our entire data said that carbon and nitrogen metabolism related genes are differentially expressed. So we do not see much change in carbon which is expected because carbon is not a thing that changes that much. But we see some changes, which is significant in terms of liquid stock infection, but nitrogen increases significantly. And the next question comes that, how could the nitrogen in sugar beet root increase? What is the cost? So as we saw in our previous slides that nitrogen metabolism related genes are highly upregulated in the sugar beet host. So the thing is that during pathogenesis, there is a high demand for nitrogen. So the host requires nitrogen for host defense. Pathogens, they require nitrogen for pathogenesis. And so basically what it tells us that there is an uptake of nitrogen by the roots. And that's why we see nitrogen metabolism related genes are highly upregulated in the sugar beet roots. And what we clearly see here that the ratio of the carbon and nitrogen is significantly decreased. So what it tells us here is that there's a shift in nitrogen-based primary metabolism, then carbon-based. And more the infection you have, more the ratio goes down. And C by N ratio is very critical in plants because it controls a big subset of genes depending upon this ratio. So the major conclusions that we can come from this study is we have identified the key candidate genes belonging to pectin lyes and polygalactic urines from Rajuktonia solanai. And these are the candidates that actually contribute to increase root rod when our solanai is closely associated with liqueuronostoc mesenteroidase in the soil and when they infect the sugar beet roots. And pathway enrichment and analysis show different pathogenesis strategies employed by eukaryotic versus prokaryote where we can see selenocompon metabolism showing up here and then here eukaryote depending upon the nature of the pathogen whether it's sugar beet or Rajuktonia solanai, you see that carbon nitrogen and metabolism-related genes are turned on more. And then increase in cell wall degraded sugars by exogenous plant cell wall degrading enzymes and total nitrogen corroborate well with root rod symptoms and nitrogen metabolism-related gene expression during pathogenesis. And then every research should lead us to another research and the candidate genes that we have identified in our case, especially the polygalactic urines and ADP, ATP carrier protein from Rajuktonia solanai or ATP synthase genes that we have identified from liqueuronostoc mesenteroidase, we can actually take this through a future RNA-based approach which is the Higgs host-induced gene silencing. And we are actually developing some constructs to target these genes through stable transformation of sugar beets and this could be a potential and alternative strategy for future disease control besides what we currently have. And I greatly acknowledge our research leader Dave and my excellent technician page and the entire team are the technicians, Josh, Josh, incredible team, without incredible team you cannot have a great research. So I'm very thankful to all of them. Thank you for your attention.