biofortification

Finding a new direction to better corn nutrition

September 20, 2021 By Lauren Hines in Research Tags: amino acids, biofortification, maize

Arabidopsis plants line the shelves of a basement room in Bond LSC. The new biofortification direction the Angelovici lab found in maize seems to also be present in arabidopsis plants. | photo by Phillip Sitter, Bond LSC

By Lauren Hines | Bond LSC

Whether it’s through kernels, cereal or chips, corn pops up everywhere in our diet, providing nutrition to countless people all over the world.

But that nourishment isn’t enough to be satisfied, especially when a staple so widespread still lacks some building blocks key to balanced nutrition.

Researchers have tried different reverse genetics approaches to making crops like corn, soybeans and rice higher in essential amino acids — building blocks for proteins something the human body can’t produce on its own.

Many approaches have yielded limited success, so the Angelovici lab at Bond LSC took a step back to examine the seed’s nutrition from a broader perspective.

“This is where…we are slightly different than other studies,” principal investigator Ruthie Angelovici said. “We are not focusing just on lysine or just on tryptophane. We’re trying to look at it as a whole to understand the mechanism of all the amino acids composition.”

Finding the mechanism that builds certain proteins means potentially improving the seed’s nutrition. Instead of focusing on specific amino acids, Angelovici’s lab looked toward manipulating a specific process at the heart of amino acid regulation. It found that the process of protein synthesis is strongly associated with certain levels of key amino acids

“It’s super surprising,” Angelovici said. “It’s like saying the factory is responsible for variation we see in some sort of a product. If that’s the case, we can now go and look at the components in this factory that are spitting out this variation.”

It might seem straightforward to genetically biofortify corn, but adding and removing proteins low or high, respectively, in essential amino acids to corn seeds didn’t work. By the next generation of maize, the plant would reset itself through proteomic rebalancing.

“Even if you completely knock out a large amount of storage protein in the seed, the seed has an intrinsic mechanism that it can reprogram, and, ultimately, it can balance those proteins in the seed, so that was very interesting but equally challenging,” said Vivek Shrestha, first author on this study and now a post-doctorate at The University of Tennessee.

Shrestha and others in the lab decided to find which proteins were responsible for the high and low composition of certain amino acids through two complementary experiments.

The first matched amino acids created in the kernel to genes related to them. When that gave a long list of genes, the second experiment whittled it down by finding associations between amino acid compositions and proteins during seed development.

This final list of genes helped Shrestha, Angelovici and others find their new direction.

“We saw a really big group of translational-related genes that seem to be highly dominant in the last piece of the [narrowing down genes process], which we wanted to distill for further inquiry,” Angelovici said. “So, basically what we’ve seen is that the translational machinery is in the heart of the regulation of the amino acid.”

This translational machinery synthesizes proteins and seems to be associated with amino acid composition variations when it wasn’t expected to. This means that there could be something else going on where researchers could manipulate to create more nutritious corn.

“From our paper, it seems that this translational machinery itself probably also has an input that we did not really put a lot of emphasis on,” Angelovici said.

Angelovici believes that this isn’t a corn-specific finding. They reported seeing it in their model plants, Arabidopsis, as well. Next is to experiment with wild mutants of maize and understand its ribosomal footprint.

“I think we had a challenge…but as we go along, we tried to solve them,” Shrestha said. “That’s what scientists are like. They take small steps, remove the challenge and then move forward.

The study, “Multiomics approach reveals a role of translational machinery in shaping maize kernel amino acid composition,” was published Aug 15 in Plant Physiology.

#IAmScience Braden Zink

January 19, 2018 By Allison Scott in #IAmScience Tags: biofortification, biology, plant science

By Allison Scott | Bond LSC

“#IAmScience because I have learned to think critically and approach scientific unknowns in a way that will prepare me for a career as a successful physician.”

Labs aren’t born in a day. Neither are researchers.

Braden Zink, a senior biology major, could tell story after story about just that. He came to Mizzou with little knowledge of university research but with the determination to get his feet wet. As a member of Ruthie Angelovici’s lab, he did both.

“I came to college completely unaware of how research worked and the kinds of problems that research scientists work to solve,” Zink said. “I joined Dr. Angelovici’s lab during her first year as an MU professor and was thrilled to have the opportunity to help get it off the ground.”

With the lab’s goal of improving sustainability and nutritional quality of seeds, Zink has been able to make great strides in plant sciences. His current project is focused on how the size of seeds relate to their metabolic profiles.

“I had to come up with a way to measure Arabidopsis seeds because they’re the size of salt grains,” Zink said. “I came up with a protocol and performed size analysis on hundreds of ecotypes. My ultimate goal is to identify a gene or several that explain the observed variation in seed size.”

Last summer, Zink took advantage of working as a full-time researcher at Bond LSC.

“My work this past summer led to the conclusion that there’s a significant negative correlation between seed size and the quantity of several amino acids,” Zink said. “In general, I discovered that bigger seeds have proportionally less amino acids.”

This information led him to a working hypothesis that metabolic adjustments other than amino acids must be responsible for seed size variation.

Zink was able to work all summer solely on his research in Bond LSC thanks to the Cherng Summer Scholars grant funded by the founders of Panda Express, who happen to be Mizzou alumni. As one of 12 recipients — making it the most competitive grant for undergraduates — Zink’s dedication to his craft was recognized in a big way.

“I was able to focus intensely on my research and was immersed in it. Over the summer I didn’t have obligations to course work, so I was really able to be all in,” Zink said. “I believe what I’ve accomplished in research will help to set me apart from other candidates as I apply to medical school this year.”

He took his findings from the summer and presented as part of the Missouri EPSCoR program, which is run by the National Science Foundation (NSF) to provide more financial resources to scientifically underfunded states.

“I presented the poster as one of around 80 Missouri scholars,” Zink said. “Included in the presenters were students at all levels below professor, so it really highlighted what up-and-coming scientists are doing.”

After the event in late August, Zink was one of 10 presenters chosen to move forward and share their work in front of a national committee of NSF scientists. As the only undergraduate student selected from the state, it was an exciting opportunity.

“It was a closed room presentation with scientists whose work I’ve been reading for a while asking me questions about my science, so it was nerve-wracking,” Zink said. “While intimidating, this was also an incredible opportunity for my work to undergo an acid-test. Having my project hold water while being evaluated by nationally recognized scientists was an experience that confirmed that the work I’m doing is both professional and meaningful.”

While his accomplishments as an undergraduate researcher speak for themselves, Zink’s next step is medical school.

“Ideally, I want to become a cardiologist,” Zink said. “I’ve shadowed Dr. Greg Flaker — a seasoned cardiologist and head of cardiac research at the University of Missouri Hospital — and the work is something I could see myself doing in my professional career. I see this as an opportunity to offer critically ill patients 10 or 15 more years of life. It is a force that drives me towards joining this field.”

Zink plans to incorporate the lessons he’s learned at Bond LSC on his path to becoming a cardiologist.

“I’ll be doing a lot of the same style of critical thinking I do now,” Zink said. “Research has helped me do things that most undergraduates don’t get to. It helps you get ahead of the ball.”

Although there are more discoveries to be made, Zink is happy to contribute what he can to get things moving in the right direction.

“I understand that the contributions I’ve made — and continue to make — will only be a drop in a massive bucket,” Zink said. “However, each drop in this bucket is necessary if it is ever to be filled.”

#IAmScience Carson Broeker

January 12, 2018 By Allison Scott in #IAmScience Tags: biofortification

By Allison Scott | Bond LSC

“#IAmScience because learning and chasing my innate curiosity about the living world with them makes me excited to work in lab each day.”

Science can be about serendipity that leads to better discoveries and opportunities that researchers don’t expect. Carson Broeker knows this all too well.

“I wasn’t always interested in research,” said Broeker, a junior biochemistry major. “I took the three science classes my high school offered, but didn’t really get into the research side of things until I came to college.”

After speaking with his undergraduate research advisor his freshman year, Broeker began looking into which professors in Bond LSC were doing work he was interested in. From there, he contacted Ruthie Angelovici and was offered a position working in her lab during his sophomore year.

Now in his second year as part of the lab, he uses his biochemistry background to work with amino acids and uncover ways to increase nutritional benefits in seeds by analyzing their genes.

Candidate genes are selected through an observational method known as a genome wide association study (GWAS), where differences in the genetic sequences of a population are compared to their respective phenotypes. In this case, it is the amino acid levels in seeds. From there, Broeker can select a gene for further study via loss of function mutation, where the gene of interest is prevented from being expressed.

Broeker’s work has been on the model plant Arabidopsis thaliana, with hope that research performed is transferable to a staple crop species.

“My job is the precursor for engineered plants that will provide more food security,” Broeker said. “I confirm that each gene of interest has lost its function.”

If both alleles of the gene he studies have transfer DNA in them, they have supposedly lost their function. Once Broeker confirms genes that have done so, he can compare the amino acid levels from the mutant seeds to the normal wild type seeds.

“From that data, we can draw conclusions to see if the knockout of my gene of interest influenced certain amino acid levels,” Broeker said.

This information can then be used to engineer plants with sufficient nutrition to sustain a healthy human diet, which can have a great impact on the world’s food supply.

While his work with genes is the main way Broeker has been exposed to research, he is always learning from those he works alongside.

“I get to talk and work with others who either have the same or more knowledge about scientific topics than I do,” Broeker said. “Learning and chasing my innate curiosity about the living world with them makes me excited to work in lab each day.”

Next year as a senior, Broeker will join Chiswili Yves Chabu’s lab in Tucker Hall. Its focus is on developmental cell-to-cell communication and tumor progression and will allow Broeker to expand his knowledge in different areas of science, all while continuing to work in the Angelovici lab in Bond LSC.

“I’m excited to get more exposure to different types of research,” Broeker said. “Interdisciplinary aspects of research are strong right now, so joining the Chabu lab is a great way to be a part of that.”

While the added work will prove challenging, Broeker is excited about what his work in both labs will contribute to his graduate school applications, which he began working on this year.

“I’ve been studying for the GRE to prepare for applications,” Broeker said. “I worked in Angelovici’s lab last summer, but am in the process of applying for Research Experiences for Undergraduates programs for next summer.”

These programs, often referred to simply as REUs, allow students to better prepare for a graduate school path. They include workshops and seminars about various components of research, as well as GRE preparation classes.

“Under the guidance of mentors, I believe that pursuing an REU would be the best for me and my future in applying for graduate school,” Broeker said.

And while he’s excited about the opportunity to better understand research and continuing to find answers to the questions he has, Broeker is largely looking forward to life after earning his degrees.

“Trying to teach others about science is very important to me,” Broeker said. “I don’t want to go to graduate school just for more classes, I want expand our collective knowledge. Teaching people is just as important as doing the research. If you can’t teach, you won’t see any results from it.”

#IAmScience Marianne Emery

November 3, 2017 By Allison Scott in #IAmScience Tags: biofortification

By Allison Scott | Bond Life Sciences Center

“#IAmScience because it leads to innovation that makes for a better world, which is an awesome thing to be a part of.”

It’s good to have a role model, and Marianne Emery has always looked up to female pioneer scientists.

One of her favorites is Barbara McClintock, a Nobel prize winning botantist who studied how the chromosomes of corn change during reproduction.

It is from women like McClintock that Emery is encouraged to always be impactful with her research and overcome obstacles with grace.

“I think it’s inspiring to see these women in positions that have typically been male-dominated,” Emery said. “You lose your confidence sometimes when things just don’t work. You’re continuously met with obstacles, but you have to keep going.”

And that she has.

Emery works in Ruthie Angelovici’s lab at Bond LSC to understand what controls protein levels in seed. She primarily spends her time on the computer working with large data sets and trying new software, but is always excited about the findings she’s able to uncover.

“I enjoy developing new skill sets every day,” Emery said. “The most important thing I’ve learned so far is how to communicate my science and how to communicate when I’m having an issue. Conveying a problem and problem-solving in general can be hard.”

Still, Emery continues to focus on improving on a daily basis. She hopes to work for a company like Monsanto after earning her Ph.D.

“I really like the business side of science,” Emery said. “Ultimately, a bigger company would be the best fit. I also really like policy and the patenting process.”

Wherever Emery ends up, though, she hopes to become like the women who pioneered science.

“Female scientists have been so inspiring to me,” Emery said. “I hope that one day I can be a leader and a role model for other young women who aspire to be involved with science.”

#IAmScience Vivek Shrestha

October 27, 2017 By Allison Scott in #IAmScience Tags: biofortification, maize

By Allison Scott | Bond Life Sciences Center

“#IAmScience because it provides me with a platform to make that which seems impossible possible.”

Agriculture is a mainstay in Nepal, where Vivek Shrestha was born and raised. He grew up in a small farming family, but he was surprised that although a significant portion of the country was involved with agriculture, food insecurity was prevalent.

“Nepal is a small, developing nation that is naturally beautiful,” Shrestha said. “Agriculture is huge, but still a lot of people are food insecure.”

Shrestha saw this need and decided to study plant sciences as an undergraduate at Tribhuvan University in Nepal. From there, he earned his master’s degree from South Dakota State University before coming to Mizzou to pursue his Ph.D.

“The overall goal of my study is to understand the genetic architecture of seed amino acid composition,” Shrestha said. “Seed amino acid composition is a complex metabolic trait and, despite having tremendous importance in biofortification efforts in seed crops, the underlying genetics are not clearly understood.”

Currently, Shrestha works in Dr. Ruthie Angelovici’s lab at Bond LSC studying the trait to better grasp its genetic breakdown.

“The {research} quality of amino acids has a tradeoff with the quantity, which makes it more challenging,” Shrestha said. “However, our research is of paramount importance because it has millions of beneficiaries.”

Shrestha’s research helps not only with food stability in places like Nepal, but also in cutting costs for the livestock feed industry in developed nations like the United States.

“Maize is a huge part of the feed industry for the United States,” Shrestha said.

This dual interest makes Shrestha’s work that much more rewarding. Although the amino acids are complex — having multiple cellular processes and interactions — the complexity gives Shrestha motivations and excitement in what he does.

“Every day is a fresh, new day for me to explore and enjoy science,” Shrestha said.

A photo worth a field of change

July 20, 2017 By Samantha Kummerer in Research Tags: biofortification

By: Samantha Kummerer | Bond LSC

When you bite into corn-on-the-cob or a burger you probably aren’t thinking about what tiny compounds are entering your body or about how they can be improved.

But scientists are.

Those tiny compounds are amino acids and serve as the building blocks of protein. They also play a major role in a recent interdisciplinary research project at the Bond Life Science Center.

Look no further than crops like corn and soybeans. While widely eaten by both livestock and people worldwide, these plants are deficient in several essential amino acids and it takes a lot for the consumer to be satisfied. Amino acids make up a large portion of human’s cells, muscles, and tissues. They are also an important part of nutrition.

“So what do you do in order to get what you need? You eat more, right?” said Ruthie Angelovici, a Bond LSC scientist. “That’s a very big problem.”

Angelovici said the solution lies in learning to manipulate amino acids to improve the quality of the seed.

Previous experiments to improve a crop’s level of amino acid have not had much success, so Angelovici decided it was time to try something different.

She decided ask if different appearances across plants, like the size of leaves or color, have any connection based on the seed it grew from.

Past research suggested that the same genes may control both seed nutrition and aspects like structure, but Angelovici’s research is unique in its combination of research on plant structural characteristics, DNA polymorphism and metabolism. A competitive seed grant of $99,690 from Bond LSC helped get this work off the ground.

She decided a few new tools and people were needed to explore this.

Learning a new language

That’s where Heather Hunt from the College of Engineering and Scott Givan in the MU Informatics Research Core Facility come in.

While Hunt has a background in bioengineering, for years she has teamed up on projects in the Plant Sciences department.

“There’s a lot of things we do as engineers that can be very useful particularly to people in plant science, particularly in terms of equipment and instrumentation, helping them develop faster methods to do things,” Hunt said.

High throughput phenotyping is one of those things. This catalogues a large number of physical features from a study group.

The team determined the research required the rapid collecting of this characteristic data from a large number of plants and multiple levels of analysis. To achieve this, the team envisioned the construction of a physical cart along with the development of hardware and software.

Hunt explained, in the past, students from a bioengineering capstone class would work on a project like this, but the teams kept running into the same problem.

“They were all talented and dedicated and hardworking but there were just things they didn’t know because they weren’t a computer engineer or a computer scientist, so they knew how to code but they didn’t necessarily have the breadth of skills someone in that area would,” Hunt explained of her past project experience.

MU engineering undergraduates, Jacob Gajewski, Yen On Chan, Nurhidayatun Anuar, and Chloe Rees, stand besides a mobile phenotyping station they designed and built. | Photo by Samantha Kummerer, Bond LSC.

This time an interdisciplinary team made up of undergraduates in mechanical, biological and computer engineering combined to figure out if a plant’s physical characteristics hint at the content of its seeds.

The team spent the last semester building a high throughput phenotyping station costing more than $10,000 from scratch. The station is on wheels can easily be moved between growth chambers. Equipped with cameras, the device photographs eight plants at once. The images capture the plant color, leaf size, and shape along with other characteristics in seconds.

But the road to this final product came with some communication challenges caused by the multiple educational backgrounds of each team member.

“Sometimes we’d have conversations where we’re talking about one thing and we’re trying to all find the phrasing that makes sense to us and we kind of just go around in a circle and then we eventually figure it out, but we get there in the end,” Hunt explained.

Both Angelovici and Hunt said when working in an interdisciplinary team, it is very similar to having to learn a new language.

“As a biologist, I think it’s very interesting to look at how engineers are thinking on things, so working with Heather, for me, was illuminating. We work very differently, we have different languages of how we think about an experiment,” Angelovici said.

Despite minor communication barriers, Hunt said the project has gone unusually smooth for her and credits the interdisciplinary team and the in-depth planning.

“All this interdisciplinary work will be our future in biology, so I think this is a great start for them and for us,” Angelovici agreed.

What’s next?

Now that the station is built, the team is taking the summer to work out kinks and begin initial data collection.

While they hope to one day evaluate crops, the current work is with multiple the model plant, Arabidopsis thaliana.

Mobile Cart — A look inside the mobile cart the team of engineering students designed and built. The machine photographs eight plants at once using four different cameras. | Photo by Samantha Kummerer. Bond LSC.

Data collection for the mock experiment is expected to start late summer. When it does, the team will take photos of many, many plants throughout their four-week life span. The images will then be analyzed for things like the shape of the leaf, the area of the rosette, color, and plant size.

“If you can determine the amino acid content in the seed and there’s a specific physical trait of the plant that it portrays, then you can tend to look at a plant and say ‘ok that comes from the bad seed,’” said bioengineering student Jacob Gajewski. “They can then modify it to where they only grow plants with the good phenotypes.”

By December, Angelovici hopes to determine if there is a connection between what the plants look like and the quality of the seeds.

If that connection is established, the next step is to figure out how the two are correlated and if the research and hypothesis are translatable to crops.

This seed funding is one of seven awarded this year at the Bond Life Sciences Center. These awards, which range from $40,000 to $100,000 in funding, foster inter-laboratory collaboration and make possible the development of pilot projects.

Ruthie Angelovici is an assistant professor in the Division of Biological Sciences, and is a researcher at Bond Life Sciences Center. She received her degrees in plant science from institutions in Israel — her B.S. and M.S. from Tel Aviv University, and her Ph.D. from the Weizmann Institute of Science in Rehovot. She was a postdoctoral fellow at the Weizmann Institute and at Michigan State University, and has been at MU since fall of 2015.

Heather Hunt is an assistant professor in the bioengineering Department at the University of Missouri. She earned her B.S. from Iowa State University and M.S. along with her Ph. D from the California Institute of Technology. She was awarded the 2010-2011 WiSE Merit Award for Excellence in Postdoctoral Research and the 2015 3M Non-tenured Faculty Award for her current biosensors research at Mizzou.

Scott Givan is the associate director of MU’s Informatics Research Core Facility. He earned his B.S. in biochemistry from Purdue University and his Ph. D in biology from the University of Oregon.