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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.

 

Katelynn Koskie #IAmScience

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Katelynn Koskie, a Ph.D candidate, works in Mannie Liscum’s lab. | Photo by Samantha Kummerer, Bond LSC

By Samantha Kummerer | Bond LSC

“#IAmScience because I want to help unravel the mysteries of nature that will improve our futures and positively impact our planet.”

Katelynn Koskie didn’t always know she loved plants. As an undergraduate, she focused on what was above her rather than what grew below her.

“I was really interested in how galaxies interact and then I started to think, ‘you know I’ve always thought plants were really, really cool,’ and I wanted something that was a little bit more down to earth,” she said.

While she was pursuing a degree in astrophysics, she took one plant biology course and fell in love. From there she signed up for grad school and has been with plants ever since.

Koskie works with a mutated plant called hyper phototrophic hypocotyl, hph. The mutation is a variation of the lab’s model plant Arabidopsis. This variation is special. It produces more seeds, bends more under light and is stronger. It’s up to Koskie to figure out why.

That answer could have a large impact on the agriculture industry. If Koskie’s findings can be applied to crop plants like maize, farmers can grow better crops.

“Maize is more complicated than Arabidopsis, but with new techniques like CRISPR/CAS9 now it might make it a little bit easier,” she said.

She plants genetically modified seeds and then waits and observes and begins again.

It is a lot of time in the growth chamber and in the dark room, hoping the research may reveal a breakthrough.

Dean Bergstrom #IAmScience

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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.

Erica Majumder #IAmScience

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Erica Majumder, a biochemistry Ph.D candidate. | photo by Morgan McOlash, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because I am endlessly curious and the world needs scientific solutions to our grand challenges.”

That is the attitude of someone who does her research with a purpose. Since the age of 14, Erica knew she wanted to pursue a degree in chemistry. Today, she uses that passion to research how anaerobic bacteria interact with uranium; essentially asking the question, “How do microbes and metals interact?”

What’s her end game? Improved health of the environment.

Sheryl Koenig #IAmScience

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Sheryl Koenig, the Grant Proposal Manager at Bond LSC. | photo by Morgan McOlash, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because I need to connect the dots. How do all the puzzle pieces fit together? Why do things do what they do? How can I apply that to other things?”

For Sheryl Koenig, science communication is an enormous part of her daily tasks. She works with researchers and scientists during the grant proposal process to translate technical scientific concepts into persuasive and relevant content. Why? So that those scientists can access the means to expand their #MizzouResearch and make exciting breakthroughs. Sheryl literally helps turn their ideas and dreams into reality! #scicomm

Marc Johnson #IAmScience

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Marc Johnson, a virology professor at Bond LSC. | photo by Morgan McOlash, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because the mysteries of the natural world aren’t going to solve themselves.”

Since the third grade, Marc Johnson never wanted to be anything else but a mad scientist. What began as experimenting with sprouting seeds and chemistry sets has blossomed into a career in virology. Specifically, he studies the “moves and countermoves” of viral components, a few hundred thousand at time! His advice for people wondering if science is for them: “If you’ve ever stayed up until 4 in the morning to finish a puzzle, you might be a scientist.”

Lloyd Sumner #IAmScience

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Lloyd Sumner, biochemistry professor and Director of the Metabolomics Center at Bond LSC. | photo by Morgan McOlash, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because I have an infinite curiosity and we have some powerful toolsets that I am confident will make a difference, not just in plant biochemistry, but in many scientific arenas.”

What change you would like to see in this world because of your research?

“I’m a technology junkie at heart. We are developing tools that can potentially advance many areas, and not just my own personal research program. I want to continue to build upon these tools and also apply them in a meaningful manner. On the plant side, I want to discover and characterize many new biochemical pathways, and use this information to make stronger, healthier and more productive plants. I also want to apply these cutting-edge tools to an ever expanding set of problems; i.e. cancer, veterinary medicine, nutrition, etc. I’m confident that every day when I get up, by the end of that day, week or month that we are making that difference.” -Lloyd Sumner

Scott Peck #IAmScience

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Scott Peck, a biochemistry professor at Bond LSC. | photo by Morgan McOlash, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because I want to discover. I want to ‘see’ – by understanding – things that others haven’t ‘seen’ before.”

Every day we make decisions based off on what we encounter in the environment. Plants do the same thing. Scott Peck, a Chicago-area native, is a biochemist who studies how plants translate information they receive about the environment (such as changes in light and temperature) into their own chemical “decisions”, also known as signal transduction. For him, it’s about making biology into a puzzle. Put the right pieces together, and you find ways to create more resistant crops or more effective antibiotics. With today’s technology and Peck’s passion for plant communication, anything could be possible.

Debbie Allen #IAmScience

Debbie Allen

Debbie Allen, the Coordinator of Graduate Initatives at Bond LSC. | photo by Morgan McOlash, Bond LSC

By Mary Jane Rogers | Bond LSC

“#IAmScience because during their journey all graduate students deserve expertise, care and advocacy from graduate coordinators.”

As Coordinator of Graduate Life Science Initiatives, Debbie Allen facilitates several activities supporting graduate recruitment, training, mentoring and career services. In other words, she’s been the “mama bear” to many life sciences graduate students over the years, and is passionate about student advocacy. To Debbie, while understanding the hard science her students study is important, supporting those students through their challenges and triumphs, and guiding them closer to their goals motivates her every day.

Pork without the Pig

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This screenshot of a supplemental video included in Genovese’s study shows cultured pork cells contracting in response to a neurotransmitter. | photo courtesy of the Nicholas Genovese

What if you could have pork without the pig? Nicholas Genovese’s cultured meat could provide a more environmentally friendly meat
By Eleanor C. Hasenbeck | MU Bond Life Sciences Center

Scientists are one step closer to that reality. For the first time, researchers in the Roberts’ lab at Bond Life Sciences Center at MU were able to create a framework to make pig skeletal muscle cells from cell cultures.

In vitro meat, also known as cultured meat or cell-cultured meat, is made up of muscle cells created from cultured stem cells.

As a visiting scholar at the University of Missouri, Nicholas Genovese mapped out pathways to successfully create the first batch of in vitro pork. Genovese also said it was the first time it was done without an animal serum, a growth agent made from animal blood.

According to Genovese, his research in the Roberts Lab was also the first time the field of in vitro meats was studied at an American university.

“I feel it’s a very meaningful way to create more environmentally sustainable meats, which is going to use fewer resources, with fewer environmental impacts and reduce need for animal suffering and slaughter while providing meats for everyone who loves meat,” Genovese said.

The research could have environmental impacts. According to the United Nation’s Food and Agriculture Organization, livestock produce 14.5 percent of all human-produced greenhouse gas emissions. Livestock grazing and feed production takes up 59 percent of the earth’s un-iced landscape. Cultured meat takes up only as much land as the laboratory or kitchen (or carnery, the term some members of the industry have coined for their facilities) it is produced in. It uses energy more efficiently. According to Genovese, three calories of energy can produce one calorie of consumable meat. The conversion factor in meat produced by an animal is much higher. According to the FAO, a cow must consume 11 calories to produce one calorie of beef for human consumption.

And while Michael Roberts, the lab’s principal investigator, is skeptical of how successful in vitro meat will be, he said the results could yield other benefits. Researchers might be able to use a similar technique as they used to create skeletal muscle tissue to make cardiac muscle tissue. Pork muscles are anatomically similar to a human’s and can be used to model treatments for regenerative muscle therapies, like replacing tissue damaged by injury or heart attacks.

“I was interested in using these cells to show that we could differentiate them into a tissue. It’d been done with human and mouse, but we’re not going to eat human and mouse,” Roberts said. “The pig is so similar in many respects to humans, that if you’re going to test out technology and regenerative medicine, the pig is really an ideal animal for doing this, particularly for heart muscle,” he added.

While you won’t find in vitro meat in the supermarket just yet, Genovese and others are working toward making cultured meats a reality for the masses. Right now, producing in vitro meat is too costly to make it economically viable. Meat is produced in small batches, and the technology needed to mass-produce it just isn’t there yet.  Genovese recently co-founded the company Memphis Meats, where he now serves as Chief Scientific Officer. The company premiered the first in vitro meatball last year, at the hefty price tag of $18,000.

“We are rapidly accelerating our process towards developments of technology that we hope will make cultured meats accessible to everyone within the not-so-distant future,” Genovese said.

Nicholas Genovese was a member of the Roberts lab in Bond LSC from 2012 to 2016. The study “Enhanced Development of Skeletal Myotubes from Porcine Induced Pluripotent Stem Cells” was recently published by the journal Scientific Reports in February 2017.