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Striking Oil

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Jay Thelen, a Bond LSC researcher, next to the GC-MS machine in his lab, which tracks seed oil measurements. | photo by Mary Jane Rogers, Bond LSC

How to get crops to produce seeds with more oil

By: MJ Rogers, Bond LSC

You can’t get blood out of a stone, but Jay Thelen wants to find ways to get more oil from seeds.

“We’re specifically working on the metabolic engineering of oil seeds. Broadly, trying to increase the oil content of crops and raise the value of the seed in the process,” said Thelen, a Bond Life Sciences Center researcher.

Seed biology and metabolic engineering have long been interests for Thelen, and his lab combines biochemistry with cutting-edge proteomics technology to identify new regulatory modules for key metabolic enzymes.

But let’s start with why seed oil is important.

Seed oil is big business, enough so that scientists are trying to maximize the amount of oil that seeds produce. It represents an important, renewable source of food and feedstocks, used for everything from salad dressing to combustible fuel. More than 448 million tons of oilseed crops were consumed in 2015-2016, according to USDA Economic Research Service.

The oil essentially comes from lipids, or fats, in plant seeds or fruits. All plant cells contain lipids, and embryonic cells within young seeds are poised to make an abundance of them, especially the storage lipid triacylglycerol.

While tree nuts can be up to 80 percent oil, most oilseed crops store 15 to 45 percent oil within their seed. Through biotechnology and metabolic engineering scientists want to increased this amount, something Thelen aims to do.

His end goal is to increase the oil in crops such as soybeans and canola, but any biotechnological idea for increasing seed oil starts in the model plant Arabidopsis.

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Modified plants that are being grown in Thelen’s lab in Bond LSC. | photo by Mary Jane Rogers, Bond LSC

“Arabidopsis is easy to transform in the lab and has a short life cycle,” he said. “We can use this plant to quickly demonstrate proof-of-principle for enhancing seed oil and then advance successful strategies to soybean, camelina, or canola.”

To increase seed oil content, Thelen’s lab works on a large protein complex called Acetyl-CoA carboxylase or ACCase – an enzyme that catalyzes the first step towards oil production.

“We made a recent breakthrough on the regulation of this complex,” Thelen said.

The proteins critical to this process are called BADC proteins – they are kin to an essential part of ACCase, but are inactive. BADC proteins significantly inhibit the activity of ACCase by mimicking its functional sibling and slowing the complex down.

Basically, BADC is a way for the plant to control and slow down the production of fatty acids. By “turning off” the BADC protein, ACCase is de-regulated and oil content in seeds significantly increases.

“We leveraged this discovery to make higher oil producing plants by simply shutting down expression of this gene family by RNA interference,” said Thelen. “Consequently this increased seed oil content quite a bit. We’re now in the process of studying gene knockouts for this family in soybean and camelina.”

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Yajin Ye, a postdoctoral researcher from China, adds water to the plants in Thelen’s lab. | photo by Mary Jane Rogers, Bond LSC

Yajin Ye, a postdoctoral researcher from China in Thelen’s lab, is in the thick of this work. He spends his time modifying seeds to maximize seed oil content and tracking the seed oil measurements in the GC-MS instrument.

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The GC-MS machine, which tracks the seed oil measurements. | photo by Mary Jane Rogers, Bond LSC

“If you want to know how much oil is in each seed, you use this instrument,” Ye said, pointing to the GC-MS. “It measures the oil content of the samples we provide it.”

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Arabidopsis is a model plant used in Thelen’s lab and the seeds are much smaller than most people would expect. | photo by Mary Jane Rogers, Bond LSC

Arabidopsis seed are much smaller than most people expect, so tiny and light that researchers have to be cautious that they don’t become airborne and cause cross-contamination. In addition, soybean plants are kept upstairs in a fifth floor greenhouse at Bond LSC. While camelina are grown in growth chambers within Schweitzer Hall as part of a collaboration with Dr. Abraham Koo, an assistant professor in the Biochemistry Department.

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Thelen’s soybean plants, which are kept in the fifth floor greenhouse of Bond LSC. | photo by Mary Jane Rogers, Bond LSC

“Each of the [soybean] plants are transgenic and were screened for higher oil content as a result of BADC gene silencing. The plants are harvested every three or four months so the seed oil content can be monitored,” said Ye.

Thelen has begun patenting the BADC technology and another strategy for engineering ACCase to “make it a more efficient enzyme complex.” The BADC technology was co-invented with his previous graduate student, Matthew Salie, who now works as a postdoctoral research associate at the Scripps Research Institute in San Diego. The patent examination process can take years, but if the technology is approved it would mean a huge influx of money in the agricultural market.

“The math is quite simple,” Thelen said. “A one percent increase in soybean seed oil translates to hundreds of millions of dollars. The numbers fluctuate depending on the market, but a one percent increase translates to about 200 million dollars for soybean alone. A five percent increase, which again, I think is achievable, when realized across the diversity of oilseed crops, we’re talking billions in added crop value annually.”

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Yajin Ye, a postdoctoral researcher from China, with Vanildo Silveria and Caludete Santa Cantarina, two visiting faculty from the State University of Rio De Janerio in Brazil. | photo by Mary Jane Rogers, Bond LSC

His innovative approach has gotten the attention of scientists and researchers all around the world. Vanildo Silveria and Claudete Santa Cantarina, two visiting faculty from the State University of Rio De Janerio in Brazil, came to Bond LSC specifically because of Thelen.

“Jay is an expert in the field and we wanted to work with him,” said Silveria.

At the moment, it seems like his research is on the right track. The initial data from Arabidopsis shows that silencing the entire BADC gene family substantially raises seed oil content, which is the main objective of his study.

“Preliminary, first-generation transgenics show soybean with higher oil. But these are greenhouse grown. Randomized field trials are still be awhile out,” Thelen said. “We’re getting closer, but still a few years away from that goal.

Thelen’s most recent oilseed research published in the journal Plant Cell in 2016, titled “A Family of Negative Regulators Targets the Committed Step of De novo Fatty Acid Biosynthesis.” 

Jay J. Thelen is a professor of biochemistry at MU and a researcher at Bond Life Sciences Center. He received degrees in both biological sciences and biochemistry – a B.S. from the University of Nebraska, Lincoln and a Ph.D from the University of Missouri, Columbia. He was a postdoctoral fellow at Michigan State University and has been at MU since 2002.

 

Megan Sheridan #IAmScience

Megan Sheridan

Megan Sheridan, a Ph.D candidate in biochemistry, works in Dr. Michael Robert’s lab. | Photo by Mary Jane Rogers, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because it’s extraordinary knowing that a small step towards a treatment could positively impact someone’s life down the road.”

Megan Sheridan doesn’t let anything slow her down.

From presenting at the Society for the Study of Reproduction’s Trainee Research Competition last week—and winning first place—to finishing up her thesis while working in Dr. Michael Roberts’ lab, she’s always juggling multiple projects. Sheridan is finishing up a Ph.D in biochemistry and hopes to graduate in December 2017 or May 2018, depending on how quickly she finishes writing her thesis.

“I was lucky enough to pick up a project studying Zika virus infections early in pregnancy,” she said. “It was one of those perfect timing moments, and we ended up getting some pretty exciting results off the bat. Now I’m really inspired by the direction my thesis work is going and find that my projects are very different but that makes things exciting.“

Sheridan’s thesis focuses on using stem cells as a model for early placenta development and how preeclampsia and viral infections like Zika impact a pregnancy. Preeclampsia is a condition during pregnancy that causes high blood pressure and protein in the urine. The disease likely occurs in the first trimester, but the symptoms don’t evolve until the 2nd or 3rd trimester. To study it, Sheridan uses stem cells generated from umbilical cords of babies born to mothers experiencing preeclampsia or a normal pregnancy, and then uses those cells to determine what defects in the placenta might contribute to the disease preeclampsia.

“I would like to learn as much as possible about the placenta and human pregnancy,” she said. “There are so many unknowns in this area of research because you can’t access the placenta during a pregnancy without disrupting the pregnancy. There are many complications that effect the mother and baby, and if more was known about normal placenta development in pregnancy, then we may be able to better understand and prevent some of those complications.”

Sheridan completed her undergraduate degree at MU, and urges undergraduates to get started in research early, as she believes it gives students a stronger foundation for graduate school. She also believes that mistakes are part of the research process, and wasn’t afraid to share one that she made early on in the Ph. D program.

“I remember in my very first rotation as a graduate student I was learning how to transfect cells with DNA so we could do a reporter assay. We were in the process of adding all the reagents, and between the student I was working with and myself we got confused about who added what,” she laughed. “Somehow, we never added the DNA- an integral part of the transfection! So a week later when we were analyzing the data, we noticed there were no values at all.”

After graduation, Sheridan hopes to experience living outside of Missouri for her postdoc placement. She’d like to stay in academia, and looks forward to continuing to research and teach.

Perhaps one day she’ll even return to MU and Bond LSC!

Kevin Kaifer #IAmScience

Kevin Kaifer

Kevin Kaifer, a Ph.D candidate who works in Dr. Christian Lorson’s lab. | Photo by Mary Jane Rogers, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because there are people suffering all over the world and this is where I’m most likely to make any kind of an impact.”

When he came to MU three years ago, Kevin Kaifer knew he wanted to work in Bond LSC. He felt it was where the best science and collaborations were happening on campus, and everything that he needed for his research – a vivarium, a DNA core, and proteomics core – were all conveniently housed here.

“I entered research because I thought the complexity of cellular life is the most fascinating topic in the world,” said Kaifer. “I wanted to be a part of it.”

He completed his undergraduate degree in biology at Truman State University and is currently part of Dr. Christian Lorson’s lab. There, Kaifer is learning transferable skills – everything from communication skills to the production of recombinant gene therapy vectors – all of which will give him a strong foundation for a career in industry.

“The growing promise of gene therapy as a safe and realistic treatment option has led to the start up of many biotech companies that are making really exciting progress,” he said. “This is where I think I will be best able to contribute to science and therapy.”

For undergraduate students who are just getting started in a science field, Kaifer emphasizes that success in science comes and goes.

“In my own personal experience, success in science only comes after a significant set of hurdles,” he said. “You have to be okay with feeling stupid, because part of your job description is to answer questions to which you do not know the answer. I would actually be concerned if you were not struggling to feel successful.”

Christopher Garner #IAmScience

Christopher Garner

Christopher Garner, Ph.D moments before his sucessful dissertation defense. | Photo by Mary Jane Rogers, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because I believe that the collective pursuit of scientific knowledge is what moves us forward as a species.”

In the time leading up to Christopher Garner’s dissertation defense, you never would have known if he was nervous. He was confident and composed, and the conference room at Bond LSC was completely filled with his professors, friends and well-wishers. Dr. Walter Gassmann gave a complimentary introduction to the dissertation, saying, “I don’t know if I’ve ever seen a student so prepared.” Needless to say, Garner passed with flying colors.

Garner completed his undergraduate degree at the University of Missouri, St. Louis. After graduating, he went to work on a small R & D team at a St. Louis company. That was his first experience with research and his mentor was influential in persuading Garner to go to graduate school. During graduate school at MU, Garner worked in Gassmann’s lab at Bond LSC, researching the inner workings of the plant immune system. His favorite part of working in the lab was constantly conducting new experiments.

“It’s really satisfying to make a prediction and then see it come true,” said Garner. “It can be equally exciting to see things are radically different than what you predicted.”

His dissertation – “Should I slay or should I grow? Transcriptional repression in the plant innate immune system” – focused on the tradeoffs between growth and defense that plants face when mounting an immune response. While the immune response is essential for the survival of plants in the face of pathogen infection, expression of defense-related genes can interfere with growth and development and must therefore be kept under tight control. His research identified a protein involved in preventing an overshoot of the immune system after it has been activated, thereby contributing new information to the field.

“If there is some way in which I can contribute to the pursuit of scientific knowledge, be it through research or teaching others about science, then I feel like I have done something worthwhile,” said Garner.

Congratulations on a successful defense, Chris!

Walter Gassmann #IAmScience

Walter Gassmann

Walter Gassmann, the new Interim Director of Bond LSC. | Photo by Roger Meissen, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because science is the best way to solve problems and help people. And the laws of nature write fascinating stories.”

Walter Gassmann, the new Interim Director of Bond LSC, has been an important part of the MU science community for more than a decade. He’s a member of the Interdisciplinary Plant Group, a researcher in Bond LSC and a professor in the Division of Plant Sciences within the College of Agriculture, Food and Natural Resources.

His research deals with how plants fight diseases and he specifically investigates the inner workings of plants’ immune systems, which are highly specialized in detecting the presence of foreign and potentially harmful organisms. Apart from figuring out how this detection works, Gassmann is interested in finding mechanisms that plants use to keep their immune system in check. The plant immune response is very potent in stopping pathogen spread, but if left unchecked it has the tendency to harm the surrounding plant tissue as well.

Fundamental plant pathology research, what Gassmann’s work deals with, has contributed to many agricultural gains, and will continue to provide avenues for improved crop yields. It has also led to many new insights for biology in general. For example, the concept of a virus was first developed in the late 1800s with work on tobacco mosaic virus. The tit-for-tat between plants and their pathogens has shaped plant immune systems and pathogen countermeasures for eons, and also affords a fascinating glimpse into the processes that shape the evolution of complex organisms.

In recognition of his outstanding contributions to plant pathology, Gassmann was elected as a Fellow of the American Association for the Advancement of Science in 2016.

We look forward to your leadership, Dr. Gassmann!

Paige Gruenke #IAmScience

Paige Gruenke

Paige Gruenke, a Ph.D candidate in Dr. Donald Burke’s lab in Bond LSC. | Photo by Mary Jane Rogers, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because I am fascinated by life on a molecular level and inspired that my research could positively impact medicine.”

As a graduate student in Donald Burke’s lab at Bond LSC, Paige Gruenke explores the role of ribonucleic acid, or RNA. That means her work involves a lot of test tubes. She looks at how specialized RNA molecules, called aptamers, bind tightly and specifically to proteins from HIV to prevent the virus from replicating. Her job is to locate the aptamers that bind to HIV proteins from a very large starting pool of RNA sequences by doing repeated cycles of removing the sequences that don’t bind and keeping the ones that do, until the strong binders dominate the population.

“A lot of the things I do don’t sound very exciting,” said Gruenke. “It’s throwing components into tubes and waiting for things to happen. It might sound mundane, but it’s all for the greater good.”

Gruenke hopes that her research will give scientists a better understanding of HIV, because understanding the virus will lead to better drug treatments and eventually, a cure. She is finishing up her second year as a Ph.D candidate in biochemistry, and plans to graduate by summer 2020. Gruenke has always been interested in the area of molecular medicine, but she has some advice for students who are just getting started.

“On a day to day basis, many experiments fail,” said Gruenke. “You’re always going to be learning something you didn’t know before. So, don’t be disheartened because something didn’t work out — just keep trying. Because whenever you have an ‘Aha!’ moment, it makes it all worthwhile.”

Kwaku Tawiah #IAmScience

Kwaku Tawiah

Kwaku Tawiah, a Ph. D candidate in biochemistry at MU, stands near his lab station in the Burke Lab in Bond LSC. | photo by Mary Jane Rogers, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because of where I come from. If you look at Africa, we have some of the most dangerous infectious diseases in the world…When you see these diseases first-hand and the havoc they cause, you want to solve the problem. People with different perspectives will make a difference in medicine.”

Growing up in Ghana gave Kwaku Tawiah a different outlook on medicine. Tawiah works in Donald Burke’s lab in the Bond LSC, and spends much of his time engineering nucleic acids and analyzing cell cultures. He hopes his research will help with early diagnosis of diseases, and wants to eventually bring it back to Ghana. He has a strong relationship with the other researchers and scientists in the Burke lab.

“In Bond LSC, and especially in my lab, it’s the people that matter,” said Tawiah. “When I came here, I knew nothing. I started with the basics and the people in my lab were patient enough to teach me the tools and skills that I needed. The people here are what keep me going.”

Tawiah said that his parents had a direct influence on both his education and career choices. Both his parents were teachers, so they were able to see his strengths and weaknesses, and saw that he was well suited for science. He completed his undergraduate degree at Lindenwood University in 2012 and is currently in his third year as a Ph.D candidate in biochemistry at MU.

For young scientists just starting off, Tawiah believes that you must be willing to learn and listen to the people around you.

“It doesn’t matter what you know, because if you’re humble you will do all right,” said Tawiah. “It’s not about what you know, but what you’re capable of knowing. If you’re not willing to learn, it’s going to be hard. Having a harmonious relationship with the people around you is key to learning.”

Shannon King #IAmScience

Shannon King

Shannon King, a Ph.D candidate in the Biochemistry department at MU. She works in Scott Peck’s lab at Bond LSC. | Photo by Mary Jane Rogers, Bond LSC.

By Mary Jane Rogers | Bond LSC

“#IAmScience because I plan to use my career to help develop agricultural innovations for the hard-working farmer.”

Most of Shannon King’s support system – her friends, grandparents, and boyfriend – are all farmers. They’re her inspiration and part of the reason her career goal is to use science to help farmers.

She’s currently a Ph.D candidate in the Biochemistry department at MU and works in Scott Peck’s lab at Bond Life Sciences Center. She’s also part of a $4.2 million grant the MU Interdisciplinary Plant Group received to fund crop research.

“I went into science because I wanted to help farmers,” King said. “With this grant, I get to go out into the field every day and be a ‘fake farmer,’ as I call it. And then I get to go into the lab and look at the science of it all. This grant gives me an everyday reminder of whom I get to help with my research and a whole new appreciation for science.”

The official name of the project is “Physiological Genomics of Maize Nodal Root Growth under Drought.” Its goal is to develop drought-tolerant corn varieties that make efficient use of available water. The project is interdisciplinary in nature and includes individuals from MU’s College of Arts and Science, School of Medicine, College of Agriculture, Food and Natural Resources, and School of Journalism. King said part of the fun of this grant is working with her team when things go wrong.

“None of us are engineers, but we’ve been doing a lot of re-engineering of our system to make things run smoothly. It’s rewarding to have all of us come together and try and make this project work.”

Jacqueline Ihnat #IAmScience

Jacqueline Ihnat

Jacqueline Ihnat, one of the 12 Cherg Summer Scholars chosen from within the Honors College at MU in 2017. | Photo by Mary Jane Rogers, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because I am able to apply what I learn in the classroom to research that makes progress towards a better future.”

Jacqueline Ihnat was recently selected as one of the 12 Cherng Summer Scholars within the Honors College at the University of Missouri. This scholarship provides her with $8,000 to fund her summer research. She’s fascinated with cells and how our bodies function, so this summer she’s studying the role of specialized stem cells in muscle regeneration and how they interact with muscle fibers—specifically the role of Eph-A3, a type of cell-surface receptor. Her faculty mentor for this project is Dawn Cornelison, an investigator in MU’s Bond Life Sciences Center.

“Doing research helps keep me focused on the bigger picture,” Ihnat said.

“Sometimes in class we learn things that don’t seem entirely relevant or useful, but being part of a research lab allows me to apply some of the knowledge that I gain in the classroom. It’s a daily reminder of why I’m learning what I am.”

Dean Bergstrom #IAmScience

Dean Bergstrom

Dean Bergstrom, the new building manager for Bond LSC. | Photo by Mary Jane Rogers, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because I provide the world class scientists of Mizzou’s Bond Life Sciences Center with the finest facilities available.”

As the new building manager for the Bond Life Sciences Center, Dean Bergstrom makes it possible for everyone else to focus on his or her research. He’s worked in Bond LSC for nine and a half years as a research technician, and in Tucker Hall eleven years before that. His unique science background and hands on knowledge of this building means that he knows exactly what scientists need to complete their projects. A facility with such diverse research interests as Bond LSC might seem overwhelming to manage, but Dean is eager to tackle the challenge.

“My background as a technician means I’m the perfect person to step into this role,” he said.