The upsides of a Falling Numbers immunoassay rapid test with Dr. Amber Hauvermale

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Resources:
New grant to help grain farmers find low falling numbers sooner

Evaluating Grain Quality and Mitigating Economic Loss with Dr. Hauvermale and Dr. Thompson

Washington Grain Commission

WSU Wheat & Small Grains website

Contact information:
Dr. Amber Hauvermale, ahauvermale@wsu.edu, 509-288-1584

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Episode transcription:

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Drew Lyon: Hello, welcome to the WSU Wheat Beat podcast. I’m your host, Drew Lyon, and I want to thank you for joining me as we explore the world of small grains production and research at Washington State University. In each episode, I speak with researchers from WSU and the USDA-ARS to provide you with insights into the latest research on wheat and barley production.

If you enjoy the WSU Wheat Beat podcast, do us a favor and subscribe on iTunes or your favorite podcast app and leave us a review so others can find the show too.

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Drew Lyon: My guest today is Dr. Amber Hauvermale. Amber is a research assistant professor in the Department of Crop and Soil Sciences at WSU with expertise in molecular biology, protein biochemistry, seed physiology, and hormone signaling. Prior to getting her Ph.D. at Washington State University, she worked in industry and has always been involved in projects occurring at the intersection between basic and applied research.

In her current role, Amber is focusing on the physiological mechanisms that contribute to the low Falling Numbers in wheat and she has been developing a new rapid immunoassay for low Falling Number detection. Hello, Amber.

Dr. Amber Hauvermale: Hi, Drew. Thanks for having me.

Drew Lyon: Always a pleasure to have you on.

Recently, you received just under $40,000 of funding from the Washington Grain Commission in fiscal year 2024, which is the current fiscal year, to support your work on the development of 21st Century alpha-amylase immunoassays–I have trouble with that word–to replace the Hagberg-Perten Falling Numbers method. What does this money allow you to do that wouldn’t get done without it?

Dr. Amber Hauvermale: Well, this money has been hugely instrumental and it was actually part of a big effort to leverage national dollars through the Foundation for Food and Agricultural Research, the FFAR program. And so, that program requires matching funding essentially from stakeholders—so, a dollar-for-dollar match. And the funding that came directly from the Washington Grain Commission was hugely important for leveraging those dollars.

Specifically, that money will go toward helping to pay salaries for myself and others on the project. It will help to buy really important resources because some of the experiments that we’re actually doing are quite expensive and would be cost prohibitive otherwise. It’s also going to be used for Extension efforts and outreach, and that’s probably one of the biggest things that that it will be used directed towards as the project progresses.

So we’re hugely grateful for the support and we wouldn’t be in this situation doing the project that we’re currently doing without it.

Drew Lyon: And I know the Washington Grain Commission likes it when their funds can be leveraged to get more dollars, which is exactly what it’s done.

Dr. Amber Hauvermale: Yes. Yes.

Drew Lyon: Yeah, very good. So how is your work on the project progressing?

Dr. Amber Hauvermale: So, just to kind of give a brief update on where we’re at, this particular project started in February of what year are we in now, 2023? So we’re about six months in into this particular project and things have been going really well. We are running to keep up.

But back in April we had our industry collaborators–those that are helping us to scale the rapid test–visit the Pacific Northwest and actually visit with the [Washington] Grain Commission and folks at WSU doing some of the research as well as the elevators to really tie together all of the pieces of the grain chain to have a better understanding of who they’re going to be working with and why it’s so vitally important that we have a test that works for this region and the grain chain at-large.

So, we are moving along really swiftly. We are in the thick of acquiring samples right now from our elevator collaborators that will then be used for the calibration of the rapid test. So, the point of the calibration is to actually have the rapid test report out in a similar fashion to the Falling Numbers test, so a metric that folks using the Falling Numbers test understand.

And you would think that it would be easy to source samples, but, you know, around here we hope that we don’t have a low Falling Numbers event because we know that that’s bad for growers. But also, we need samples that have problems with Falling Numbers in order to do the calibration. So, it’s been a really interesting logistic challenge to get those things to come together. But things are moving along really well.

And I’m also happy to report that we have a new graduate student who started this summer. His name is Jack Kelly. I know he’s really interested in the future to do one of these episodes with you. He’s going to be working with me on some of the proteomic work and some of the biochemistry work to better understand the physiology of LMA and PHS that cause low Falling Numbers in wheat.

Drew Lyon: Okay. So, I’m just kind of curious–you’re working on this rapid test, an immunoassay–how is that different? And I’m kind of curious–that test you say reports back kind of like the Falling Number, so it’s going to be more–when I think of immunoassay, I think of the little line on the COVID test or something, right? You have it or you don’t have it. But will this test be able to tell you whether you’re Falling Number is above 300 and below 300 or just how much it is?

Dr. Amber Hauvermale: Yeah. Your question is a great one. So, just to give it a brief definition about what an immunoassay is–an immunoassay is just a procedure that detects a specific target using antibodies, and it’s different than a Falling Numbers test. So, the way the Falling Numbers test works is to indirectly measure starch degradation due to an enzyme called alpha-amylase.

So, for those that have heard us talk about it before, it is affectionately referred to as the gravy test. So, in a Falling Numbers test, you just mix water and wheat meal together and then you heat it to form a gravy. You drop a weight through it, and starch that has a lot of integrity is thick. And so, it takes a long time for that weight fall through, whereas starch that’s been broken down by alpha-amylase has less integrity and so that weight falls through very quickly. And so, with the Falling Numbers test, you get an indication of starch integrity, but you don’t really get a sense for how much alpha-amylase is there.

With an immunoassay, on the other hand, we’re able to quantify the amount of alpha-amylase that leads to, say, a Falling Number below 300 or how much is there for a 300. And so, what we’re trying to do because the industry, all the way through the grain chain, is used to Falling Numbers report number of seconds, that you know, a 300-second as you referred to–what we’re doing is we are calibrating the rapid tests to samples of known Falling Numbers so that we have a quantifiable measure of alpha-amylase associated with a Falling Number of 300 or 275 or 250.

And you asked about whether or not this is just simply going to be a yes or no answer with the new test that we’re developing, and actually what’s nice about immunoassays is that they can be developed in multiple ways. So, at the most simplest level, a dipstick. So, what you talked about a COVID test, so you get kind of a color metric report based on the amount of the target protein that’s there. It’s either absent or it’s really red.

You can also develop these tests to quantify or be more high throughput so you can develop a 96-well platform. And so, what we’re envisioning is that depending on where they will be used, will determine the format. So, for example, at an elevator, when they are trying to put the grain away or bin the grain, you know, there’s not a whole lot of time to have a fancy laboratory 96-well platform. You know, the amount of time that it takes from when the truck arrives to the scale to when the truck is offloading at the pit is maybe only five minutes. So having a lot of equipment or really fiddly equipment isn’t useful at the elevator. So, having a dipstick or something that’s small that can be run quickly is, because, you know, there they are just simply asking, yes or no, do we have a problem or don’t we?

The breeding side, on the other hand–so the breeders, you know, as they’re breeding for resistance to the causes of low Falling Numbers, they may want to know more about physiology or they may want to implement this early in their breeding programs before they’re at the stage of trialing varieties out in the field.

So, they may have thousands of lines that they’re looking to get a sense of, and so dipstick tests may not be the most practical way to screen 5000 lines. So, in that case, you know, having an immunoassay that’s higher throughput like a 96-well version or something like that is going to be more efficient and useful in those sorts of applications.

Drew Lyon: Okay. When you talk about a 96-cell, for those who might not know what that is, what is that?

Dr. Amber Hauvermale: What is that? It’s just a 96-well plate. So, it’s a plate with 96 little micro-wells that you can do an individual reaction in each one of those. It’s really common to use that sort of setup for DNA and RNA extractions. And it’s kind of one of the common tools in our tool chest in the laboratory.

Drew Lyon: Yeah, in the laboratory. But you’re not going to find them out and about.

Dr. Amber Hauvermale: No, it’s definitely not a field application.

Drew Lyon: Okay. So, when or why did the effort to develop this new test begin?

Dr. Amber Hauvermale: Oh, that’s a great story. And yeah, it’s had quite the history and a lot of people involved pushing towards this effort. And just to kind of give you a background, a little bit of background about myself, so I started on the project in about 2018 and at that point a lot of people had already been pushing for rapid tests for several years.

And recently in a series of interviews with critical folks in the grain chain and critical stakeholders, I was meeting with Alex McGregor–actually my colleague Alison Thompson and I were–and he gifted us a stack of communications from the very beginning of pushing for this effort, so in 2016.

In 2016 there was a very, very large Falling Numbers event that impacted the Pacific Northwest, and unlike previous years or, you know, many of the years since, it happened everywhere. So there really wasn’t enough grain, sound grain in the system–in the grain chain–to counteract all of the low Falling Numbers. And so, a lot of folks–a lot of farmers were impacted very negatively because they were docked a lot. I think there were, it’s estimated, anywhere from $30-100 million in losses in the Pacific Northwest in 2016.

So, it was then that folks like Alex and the late Craig Morris and Jim Moyer kind of put their heads together and said, “okay, we need a better way to evaluate the quality of the grain at receival stations.” And that effort has been moving forward ever since.

Drew Lyon: Okay. So, you’ve talked about it a little bit, but what were some of the challenges and the benefits of a new test like this?

Dr. Amber Hauvermale: So, I think that some of the challenges with any new technology is just dealing with the normal logistical barriers that come up, right? So, I spoke about one a little bit earlier, you know, just getting samples to people that actually, for the purposes of calibration, that have been negatively impacted by a Falling Number event. You know, the past couple of years in the Pacific Northwest, it’s been pretty quiet. So, you know, making sure that we’re connected with not only, you know, people in our immediate region, but across the United States to help us with that effort.

And, you know, some of the other challenges are just with educating people about how the test works, what its benefits are, what its strengths and weaknesses are, and then with, you know, having people who are interested and actually willing to validate a new test. You know, early adopters. And some people are really excited to try new things and some people are more cautious because, you know, there’s been a lot of new technology that’s come on the market and sometimes it performs really well and sometimes it doesn’t perform well.

So, the benefits of the development of a new rapid test are hopefully that if it performs the way that the industry needs it to, it will be able to allow receival stations to start segregating grain, hopefully in real time. And hopefully it’ll provide an early warning for folks, you know, if there are problems, and then hopefully it can be used not as a post-harvest management tool but also as a pre-harvest management tool–so, something that can be utilized by the breeders to help develop new lines that are resistant.

Drew Lyon: Okay. You’ve spoken about some of the stakeholders in this business, I guess, who might be affected by this test. It sounds to me like it could be quite a large group interested in this–who do you see belonging in that group and do they all support it or are there some naysayers in there?

Dr. Amber Hauvermale: Yeah. From the perspective of the project itself, I think anybody that’s in the grain chain is a potential stakeholder for this project. So, you know, we’re talking about researchers, we’re talking about elevator workers, we’re talking about bakers, millers, and folks that are, you know, at the export terminals.

And when speaking with people, you know, the most eminent folks that–like the point of first need seems to be with management at the elevators. And we typically have people that fall into two groups. When I’ve been speaking to different groups within the grain chain, and those are people everyone has said, “yes, we need a new test.” But we have those that are early adopters and those that are more cautious that want to see how it performs, and both are valid.

So as a strategy with this project moving forward, what we have done is we have strategically partnered with folks that have identified themselves as early adopters and they are at the elevators, they’re in research, they are doing Extension, they are like at the Wheat Marketing Center doing education, you know. So, we have key people at every link in the chain essentially, who will be testing the test in real time, providing feedback about how it can be improved and also who’s working with the industry at-large to provide opportunities for Extension and feedback and ways to improve methodology moving forward.

Drew Lyon: Okay, so what do you see as some of the barriers and/or benefits to adoption of this new test?

Dr. Amber Hauvermale: I think some of the biggest barriers that have been reported to us are that because of the way that the grain, specifically in the Pacific Northwest, moves for export, it’s our export industry stakeholders that may not be early adopters of this. So, from the standpoint of the export terminals and those that are contracting with U.S. wheat associates for Pacific Northwest white wheat, you know, the industry standard is a falling number of 300 seconds or better.

But people have been really, really open about what it would take for adoption. And so, we know that we have to have FGIS standards and we know that we have to have AACC approved methodology and we have been told that, you know, with time it is likely that our biggest naysayers, if you will, are people that are used to the old technology, once they see performance, will likely adopt.

And then, the benefits for adoption have been, you know, we have folks at the local elevators that are like itching to get their hands on something that they can use in the heat of harvest to help them better manage problems early on at point of delivery, and then all the way through the grain chain because while it’s easy to get a Falling Number report back early on in harvest, as harvest progresses and you have more and more trucks that line up that elevators or you’re just trying to move that train load or that truck load down to the port, it gets really busy and really hectic. And so, if there is a way that takes less time, I think that the elevators are really, really eager to try this out and to use it.

I think that it will also, from the standpoint of research–basic research, hopefully give us a better idea of how this enzyme alpha-amylase is regulated during germination and during weather events that cause high levels of expression. And then that information can then be used as part of the breeding efforts to prevent it from happening in the future.

Drew Lyon: Alright. So, these elevator operators who are just itching to get their hands on it, when are they going to get their hands on it?

Dr. Amber Hauvermale: That’s a great question. So actually, our industry partner right now is in their feasibility phase. And like I said, this field season, they are in the process of calibrating the rapid test that is being developed currently. We are hoping to see something this fall. So, what we are doing, you know, harvest will be over by the time that we have something in hand, but we are all hoarding samples, so we can through the winter actually evaluate performance so that by the time our next field season comes around, we’ll have something in hand that can be used in real time.

Drew Lyon: Alright. Excellent. So not too far off.

Dr. Amber Hauvermale: No, no, it is quickly approaching. Yes.

Drew Lyon: Okay. Well, I look forward to having you back on the program next year where you can tell us how it’s all going, because I think this is a very important topic for many of our growers and the industry in eastern Washington. So, thanks for working on it and thanks for sharing your information with us today.

Dr. Amber Hauvermale: Absolutely. And, you know, hopefully in the future we can also have some of the other folks on for updates about the FFAR project and some of their key roles. And yeah, we’re happy to do this.

Drew Lyon: Okay. And is there going to be a place where people can go to find out what’s happening? Like is the FFAR project going to have a website or a web page somewhere?

Dr. Amber Hauvermale: Yeah, so what we’re doing right now is, we are in the process of trying to coordinate that effort. A lot of the information that will be acquired through the project will be shared on public websites. Right now, the strategy is through like the Small Grains website at WSU and through our stakeholder websites—so Highline Grain Growers and McGregor Company, and there will also be an effort with the Washington Grain Commission also to share information as well.

Drew Lyon: Okay, we’ll try to get that information into the show notes when we have it.

Dr. Amber Hauvermale: Great.

Drew Lyon: Alright. Thanks, Amber.

Dr. Amber Hauvermale: Thanks, Drew.

Drew Lyon:

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Thanks for joining us and listening to the WSU Wheat Beat podcast. If you like what you hear don’t forget to subscribe and leave a review on iTunes or your favorite podcast app. If you have questions or topics you’d like to hear on future episodes, please email me at drew.lyon — that’s lyon@wsu.edu — (drew.lyon@wsu.edu). You can find us online at smallgrains.wsu.edu and on Facebook and Twitter [X] @WSUSmallGrains. The WSU Wheat Beat podcast is a production of CAHNRS Communications and the College of Agricultural, Human, and Natural Resource Sciences at Washington State University.

I’m Drew Lyon, we’ll see you next time.

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The views, thoughts, and opinions expressed by guests of this podcast are their own and does not imply Washington State University’s endorsement.