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A step into summer research

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Jacqueline Ihnat, one of the 12 Cherng Summer Scholars, outside Dr. Cornelison’s lab at the Bond Life Sciences Center. | Photo by Mary Jane Rogers, Bond LSC

Sometimes the most learning occurs outside of the classroom.

For Jacqueline Ihnat, an opportunity to pursue research at the Bond Life Sciences Center this summer will give her that chance. She recently became one of 12 Cherng Summer Scholars, a full-time, ten-week program within the Honors College at MU.

“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.”

Jacqueline Ihnat’s passion for science started in high school. Her high school biology teacher ignited that love by teaching her how to struggle through difficult problems and concepts. Now, Ihnat is an MU pre-med student with a major in business management and a minor in Spanish.

Since Ihnat is fascinated with cells and how our bodies function, she’ll be 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. This project will take place in the lab of Dawn Cornelison, a Bond LSC biologist who will be mentoring Ihnat this summer.

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Jacqueline Ihnat as she pipettes samples in Dr. Cornelison’s lab in Bond LSC. | photo by Mary Jane Rogers, Bond LSC

Each muscle in the body is unique in its length, fiber organization and fiber type patterning, so Ihnat hopes to explore why two types of muscle — fast and slow twitch muscle — develop and regenerate to maintain each specific muscle fiber-type composition.

When a muscle is damaged from exercise or injury, a muscle’s stem cells, or “satellite cells,” will multiply, move towards the injury and form new muscle, replacing the damaged fiber. There is no research that determines if “fast” satellite cells create fast fibers and if “slow” satellite cells create slow fibers, and Ihnat hopes to tackle that question this summer. This kind of research gives scientists a deeper understanding of degenerative muscle diseases such as ALS, which could lead to more effective treatments and therapies.

The Cherng Summer Scholars program is supported by a gift from Andrew and Peggy Cherng and the Panda Charitable Foundation. The Cherng’s are the founders of Panda Express, a well-known restaurant chain. These scholarships support individually designed theoretical research, applied research or artistry projects under the mentorship of an MU faculty member.

For young scientists who are just starting to conduct research and struggling to feel successful, Ihnat has a few words of motivation.

“My high school biology teacher always said that research is 30 years of frustration and disappointment followed by 30 seconds of elation when you finally make a breakthrough,” she said. “Patience is key.”

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.

Growing a more nourishing future

Nga Nguyen hopes to apply her research to increase nutrient contents in crop plants
By Eleanor C. Hasenbeck | Bond LSC

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Nga Nguyen, a doctoral candidate in MU’s Division of Plant Sciences, observes samples of a model plant species, Arabidopsis thaliana, in the Mendoza-Cózatl lab at Bond Life Sciences Center on Feb. 7, 2017. | photo by Eleanor C. Hasenbeck, Bond LSC

Plants smell better than animals, at least to Nga Nguyen. That’s one reason why she decided to study them.

“In my undergrad, I studied horticulture,” Nguyen said. “For that you don’t really learn the inside mechanisms of plants, so I decided besides knowing the cultivation techniques, I’d like to also learn about the molecular biology.”

As a fifth year doctoral candidate in the Mendoza-Cózatl lab at Bond Life Sciences Center, she hopes to combine her undergraduate background with her present research in the microbiology of plants to improve the crops of the future.

Nguyen studies how transporter proteins move micronutrients like iron through plants. By understanding how plants move these nutrients in model plants, researchers hope to apply the same understanding and techniques to crops like soy and common beans. Increasing the micronutrient content of these crops could be a useful tool in combatting nutrient deficiencies in areas where people don’t have access to meat and dairy.

But Nguyen says the benefits of studying plants don’t end there. “I hope people pay attention to plant research and study,” Nguyen said.  “If you think about it, it’s not just our food, but our clothing and the materials we use.”

#Sciku: Science and wordplay

This collection of Science Haikus were inspired by the #sciku Twitter campaign by Popular Science, which highlighted a few science haikus from readers. As a response, I rallied some of the scientists at the Bond Life Sciences Center to come up with haikus relating to their discipline of science.

The response was wonderful — Dr. Burke, of the immunology and virology core sent me a whopping 26 haikus — and many more scientists participated than I originally anticipated.

A haiku is a rigid form. It’s also a famously concise rhetoric, forcing the author to condense their meaning into the designated number of syllables (not always an easy task).

The funding environment in sciences is becoming tighter and tighter all of the time, too. There’s less money to go around and scientist are more than ever being asked to condense their grant proposals down to two or three pages (a contrast to the 15 page proposals of the 1990s).

Could it only be a matter of time until the National Institutes of Health will be giving money to research based on haiku proposals?

 

The only thing you need to read about Ebola today: An expert Q&A

Jingwio Yu, a graduate studemt, does cell surface staining in Shan-Lu Liu's virology lab. The staining illuminates cell marker expressions in experiments that deduce how viruses spread once they are contracted. | Paige Blankenbuehler

Jingyou Yu, a graduate student, does cell surface staining in Shan-Lu Liu’s virology lab. The staining illuminates cell marker expressions in experiments that deduce how viruses spread once they are contracted. | Paige Blankenbuehler

News headlines seem to feverishly spread as if they were a pandemic of the brain.

Ebola hemorrhagic fever has been the most talked about disease of the year, appearing in thousands of headlines across the world since May. Through the noise of misinformation and sensationalism, fundamental information about the pandemic becomes harder to distinguish.

In an interview with Decoding Science on Tuesday, Shan-Lu Liu, MD, PhD, a Bond Life Sciences Center investigator who studies Ebola, weighed in on the latest news.

Liu, also an associate professor in the MU School of Medicine’s Department of Molecular Microbiology and Immunology, and his lab are particularly interested in the early behaviors of the virus in transmission and how it can navigate around the host immune response.

Shan-Lu Liu, Bond Life Sciences scientists and associate professor in the MU School of Medicine department of molecular microbiology and immunology.

Shan-Lu Liu, Bond Life Sciences scientists and associate professor in the MU School of Medicine department of molecular microbiology and immunology.

Q: Talk about the transmission. Ebola doesn’t spread through air, but how easily can it be transmitted through fluids?
A: It’s hard to say. It’s really not like: touch an infected person and you got it. I don’t see that could happen so easily. As an RNA virus, it’s not that stable outside of the body, unlike hepatitis B virus (HBV) where you need to boil the virus for 10 minutes and it becomes not infectious. Because Ebola is not that stable, that should not be the reason why it’s so efficient to transmit.
I think the transmission is one of the biggest things it’s, you know, I don’t think we have a complete  understanding. We do know that it spreads by contact through body fluids and many people don’t realize that the handling of the deceased — that’s very dangerous. Touching broken skin or mucous membranes like the nose and mouth is dangerous.

Q: Talk about the incubation period and how that relates to symptoms and spreading of the virus.
A: The incubation time is 2-21 days. At first, the person will have flu-like symptoms, so you know, that’s why it’s hard to notice in the early stages. Some doctors or nurses say ‘just give him antibiotics send him home.’ But in stage two, you get the hemorrhage and it gets serious. The mortality rate is high, from 50 to 90 percent.
I think the fatality is definitely related to the late stages of the disease, especially with the hemorrhaging fever. The early stages are almost unnoticeable but that’s the time transmission might spread easier through contact with an infected person’s fluid. Before symptoms, the virus doesn’t spread.

Q: The Centers for Disease Control and Prevention said the virus could infect 1.4 million people in West Africa. Is this a realistic expectation?
A: You know, it could happen. It’s a prediction again, right? But I think the agency and the scientific community need to look at this prediction very carefully. What could be done in terms to prevent this from happening? It’s alarming.

Q: Last week, an article seemed to contradict with the CDC estimate. The headline: Some good news about Ebola: It won’t spread nearly as fast as other epidemics. What do you make of that?
I don’t know, it’s hard for me to make a comment. Nobody knows. Things can always change. We didn’t expect to see a diagnosis in the United States — like this you know, this patient from Liberia was able to travel on a plane from virus country. Who can expect that? Anything can happen. There seem to have been some mishaps because he came from that area, right? Communication is more important now but it’s hard to predict because anything could happen.

Q: How has the Ebola virus behaved in previous outbreaks?
A: The first outbreak was in 1976 in Sudan and Congo — (Democratic Republic of Congo, known as Zaire at the time). It was from contaminated needles in a hospital and originally came from fruit bats — they are one of those animals that could transmit Ebola from animals to humans. The fruit bats transmitted the virus to primates, primates transmit to humans. It’s hard to notice in the early stages.
Editor’s note: The 1976 outbreak was the first occurrence of Ebola in humans. The outbreak affected one village, infecting 318 people that resulted in 280 deaths.

Q: Much of the media has reported a vaccine for ebola was delayed. How could this happen?
A: Drugs and vaccines are a little different. The Ebola vaccine was delayed, that’s for sure. That’s because, the vaccine on trial has to go through tedious steps to get approval and so thats why when this outbreak occurs the NIH (National Institutes of Health) decides to go ahead quickly. One of the things for ebola vaccine is um, the pharmaceutical companies and the industries are not interested in developing vaccines. Do you know why? It is not a big market. Only a hundred — or a thousand or more — people will be infected by ebola, unlike other vaccines like the HPV vaccination where 200 million people need it. The companies are not interested in developing it, because there’s no money in it.
A company needs to spend a lot of money to develop a vaccine, but they don’t see the market — the market can’t do it. But somebody needs to do it. Imagine if, if the virus spread like this, you know, unpredictable, it could be worse. In terms of therapy, the drugs and antibodies, we know they are really effective. And they are specific, so they can reach the market effectively.

Q: Will a drug be enough to prevent wide spreading of Ebola?
I  think the companies and governments are speeding up to make those available. To see this prediction (the CDC 1.4 million estimate), they have to be prepared. People have put increasing attention on antibodies because a vaccine is not in the near future. So what’s the approach? A “therapeutic vaccine.” The so-called therapeutic vaccine is an antibody so you engineer, you use you know, molecular engineering technique to generate those antibodies  and they can neutralize and block viral infection. It’s more realistic for Ebola and even for HIV. The HIV vaccine has failed so many times. So that’s why I think one of the new approaches is to use a new broad neutralizing antibody.

Q: Does Ebola stay in the body, like chicken pox?
A: Ebola do not cause latent infection. HIV can become latent and become chronic. So influenza virus, ebola viral infection and others normally do not lead to latency. I think for Ebola — for this type of infection — once you block the patient and clear the virus it should be good.

Q: Has the media done a good job in educating the public?
I think in terms of news coverage they are pretty careful. I looked at the news conference by the CDC director and by those doctors in Dallas, and when they make statements they are careful not to exaggerate and also give very cautious measurements. The news media need to be aware of the danger of the virus. In the meantime, you have to be aware of the possibility of being affected.
Again, I think it is a very important problem. It’s important to let the public know the situation. If you see people who have recently traveled from those West African countries, you have to be cautious — air travel is so common. But I think the media have generally done a good job.

Q: Has the government done a good job keeping the pandemic under control?
I don’t know what they do. The air travel is a problem. Intensified screening process, that should definitely be done. It’s very bad for people from the outbreak area, and I just hope that this community won’t be affected.
To control, they should be careful. A person with any sign of the disease — they need to be quickly monitored and treated.

Q: What’s the most important take-away message for the public?
A: I think it’s an important problem and we need to solve it urgently. I hope this outbreak will teach us a lesson in terms of how important emerging infectious viruses are as it comes and goes is to public health. Based on literature and reports, if people do not have obvious symptoms, they do not produce an infectious virus. The incubation time has a big range but again, we are still trying to understand the process better. Infection is a complex process. We need to better understand the viral transmission so I think for now, we need to be very cautious.

Liu and his lab do not work with the contagious Ebola virus on University of Missouri campus. All of the studies involve use of a recombinant or pseudotyped Ebola virus which is not infectious.

At the Bond LSC, the wall wears the plants

The unusual red color of the Lobelias leaves stand out among 200 other species that thrive in the 20-foot plant wall at the Bond Life Sciences Center | Paige Blankenbuehler

The unusual red color of the Lobelias leaves make them stand out among 200 other species that thrive in the 20-foot plant wall at the Bond Life Sciences Center | Paige Blankenbuehler

Story by Madison Knapp | Bond Life Sciences summer intern

A hidden treasure on the University of Missouri’s campus is a living and breathing work of art.

In the Christopher S. Bond Life Sciences Center, a 20-foot plant wall stands as a towering tribute to the diversity of plant life and coexistence of species — a botanical landscape of more than 200 individual plants from 40 to 50 species including many ferns, vines, perennials, orchids and geraniums.

The display continues to be a visual reminder of the building’s interdisciplinary nature – much like the plants within the wall, the Bond Life Sciences Center facilitates the coexistence and collaboration between an array of researchers within its walls.

One plant cascades down the wall and stands out more than most — it looks more like a large insect than any type of plant typically seen in such displays.

Tillandsia is a genus of succulent plant native to Central America that can undergo long dry spells. The wiry plant thrives on the stone beside the plant wall without the help of a pot of soil, and is seemingly absent an essential, anatomical feature of most flora — roots.

With leaves more like Medusa's hair, Thallandsia are rootless plants mounted on stone  alongside the plant wall. | Paige Blankenbuehler

With leaves more like Medusa’s hair, Thallandsia are rootless plants mounted on stone alongside the plant wall. | Paige Blankenbuehler

They still have tiny root protrusions, but the mass is minuscule compared to the thick leaves, which take up and store water and nutrients, rather than the root system which handles that work in most plants.

Approximately 60 Tillandsias pepper the plant wall, including a few mounted directly onto the stone wall with a special kind of non-toxic rubber cement manufactured by Davis Farms based out of San Diego.

The strange plant was added to the green tapestry by Jason Fenton, an office support associate at the Bond Life Sciences Center. He first noticed the rootless plants in Belize while on a trip there in 2003 and had been growing them at his home.

“I think they’re really beautiful and distinctive,” Fenton said. “They help give variety to the wall.”

The rootless plants require special treatment — just a little extra attention from facilities manager, Jim Bixby, who waters the Tillandsias with a spray bottle once a week.

The wall started with a vision from Jack Schultz, director of the Bond Life Sciences Center. The execution and maintenance of the feature is credited to Bixby, who has honed the project since 2010.

After trying several watering systems, Bixby found one that worked. He hung rows of pots with modified flat sides and tailored a simple, drip irrigation system made for easy watering along a grid.

Within the diverse landscape and after additions of a variety of plants over the years, Bixby has seen competition between several of the species.

The wall favors the ferns, which have taken over much of the space, crowding out other plants for the eastern sunlight that flows through the windows, Bixby said.

Several long-stemmed species have found ways to cope, however; venturing out between the ferns’ curtain-like fronds to get their fair share of sunlight.

Supervising editor: Paige Blankenbuehler

The 20-foot tall plant wall outside of Monsanto Auditorium in the Bond Life Sciences Center is a nod to coexistence and diversity. | Paige Blankenbuehler

The 20-foot tall plant wall outside of Monsanto Auditorium in the Bond Life Sciences Center is a nod to coexistence and diversity. | Paige Blankenbuehler

A veterinarian abroad: Rwanda

While summer brings a slower pace for many researchers, others use it as an opportunity to learn for their profession and network with others in their field.

Bond LSC researcher Cheryl Rosenfeld recently traveled to Africa to further her learning as a veterinarian. This continuing education gives her the opportunity to learn the newest techniques in the field and network with others to learn what’s current and on the collective minds in veterinary medicine.  Through the North American Veterinary Community (NAVC), Rosenfeld has now gone on three expeditions where participants observe animals in their natural, exotic environments, attend nightly lectures and learn more about the humans near these animals. 

Previous expeditions led Rosenfeld to the Galapagos Islands and the Florida Keys, but her June 2014 trip started in Rwanda and ended in Tanzania. Here’s the first of two entries where Rosenfeld shares here experience. 

By Cheryl Rosenfeld

The fate of animal populations is generally intertwined with the predicament of humans in the area. Nowhere is this truer than in Rwanda. Most people know Rwanda for Dian Fossey’s work with the mountain gorillas and the genocide of more than 1 million Hutus and Tutsis that happened 20 years ago in 1994. In this 100-day period, an average of six individuals were killed per minute. Children that survived were often orphaned and many surviving women suffered being raped and exposed to HIV infection. In all, many still require extensive medical and psychological care. On our flight and checking into our hotel was a medical team from Harvard Medical Center that was there as part of the Clinton Foundation to assist in the medical needs.

We saw the history that continues to shape the country when we first visited a genocide memorial site just outside of Kigali where thousands of individuals were brutally murdered and the Kigali Genocide Museum that was partially funded by an English Jewish Holocaust survivor. The history of the conflict is rooted partially in Western influence that infused a social division. Prior to Europeans colonization, Hutus and Tutsis lived in relative peace and individuals could go back and forth between these two groups. The original difference was that Tutsi individuals owned more than 10 cows. The differential treatment and classification adopted by Europeans began to trigger conflict between the two groups. Prior indicators, including extensive propaganda, were ignored by the United States and United Nations. The museum includes two stained glass windows that depict the evidence that genocide was imminent and failure by other nations to prevent this tragedy. Genocide isn’t unique to Rwanda, though, and the displays describe the commonalities on their sad origins in other countries throughout history.  Outside the museum, there are several mass graves where fresh flowers are placed on a routine basis.

I was originally hesitant about traveling to Rwanda because of this history, but am very glad I took the chance. The Rwandan government has worked hard to turn around and instill pride in the country. Their economy is one of the fastest growing in Africa with construction of new businesses and hotels in Kigali. Moreover, the government has placed a ban on plastic bags and hired teams of individuals to keep the country clean. One Saturday a month, all Rwandans, including the President, are expected to participate in clean-up day, which becomes a convivial social event. While there is still sadness in the eyes of many individuals I met, I also saw hope of something better, which was inspiring to witness.

We were soon off to learn about the mountain gorillas that are now the pride of the country. During Dian Fossey’s time, she battled to prevent poaching of these magnificent and intelligent creatures. The country now realizes the worth of preserving and propagating the mountain gorilla populations. In a reasonable and safe way, they developed a tourist industry to view the various troops of gorillas. It currently costs $750 to spend one hour with the mountain gorillas. The government has restricted access to prevent gorilla habituation and stress from too many tourists.

We spent two days with different troops. While waiting in the morning to find out which troop we were responsible for trekking, we were entertained by local dancers. I regrettably made the mistake of indicating I felt fit to track the one of ten groups that was at the furthest distance.

The group that was involved in trekking the first day was called “Snow” in Kinyarwanda. I believe they received this name because they inevitably reside high in the mountains, which used to have snow. As we set off on our hike, many children came out to say “MooRahHoh”- hello in Kinyarwanda and asked for us to take their picture. We were informed that we should easily return before lunch, and therefore were only provided a package of peanuts. Unfortunately, it took us longer to hike through the forests that transitioned from bamboo to masses of stinging nettles and did not return to the hotel until 6:30 p.m. After more than three hours of hiking and our eyes finally fell upon our first mountain gorilla, the silverback of the group. Even knowing that this was the ultimate goal, we were not prepared for this amazing experience of being so close to a creature in the wild that resembled us.

We had the opportunity to meet the rest of the troop, including several 3 to 4 month old babies that were quite entertaining. The enclosed photos and videos only provide a sliver of the spectacle that we were privileged to be part of these two days of gorilla trekking that made our hunger and continued burning sensation on our face and legs from stinging nettles well worth it.

Frogs help researchers find genetic mechanism for mildew susceptibility in grapevine

Powdery mildew on a cabernet sauvignon grapevine leaf. | USDA Grape genetics publications and research

Powdery mildew on a cabernet sauvignon grapevine leaf. | USDA Grape genetics publications and research

A princess kisses a frog and it turns into a prince, but when a scientist uses a frog to find out more information about a grapevine disease, it turns into the perfect tool narrowing in on the cause of crop loss of Vitis vinifera, the world’s favorite connoisseur wine-producing varietal.

MU researchers recently published a study that uncovered a specific gene in the Vitis vinifera varietal Cabernet Sauvingon, that contributes to its susceptibility to a widespread plant disease, powdery mildew. They studied the biological role of the gene by “incubating” it in unfertilized frog eggs.

The study, funded by USDA National Institute of Food and Agriculture grants, was lead by Walter Gassmann, an investigator at the Bond Life Sciences Center and University of Missouri professor in the division of plant sciences.

The findings show one way that Vitis vinifera is genetically unable to combat the pathogen that causes powdery mildew.

Gassmann said isolating the genes that determine susceptibility could lead to developing immunities for different varietals and other crop plants and contribute to general scientific knowledge of grapevine, which has not been studied on the molecular level to the extent of many other plants.

The grapevine genome is largely unknown.

“Not much is known about the way grapevine supports the growth of the powdery mildew disease, but what we’ve provided is a reasonable hypothesis for what’s going on here and why Cabernet Sauvingon could be susceptible to this pathogen,” Gassmann said.

The research opens the door for discussion on genetically modifying grapevine varietals.

Theoretically, Gassmann said, the grapevine could be modified to prevent susceptibility and would keep the character of the wine intact — a benefit of genetic modification over crossbreeding, which increases immunity over a lengthy process but can diminish character and affect taste of the wine.

Grapevine under attack

Gassmann’s recent research found a link between nitrate transporters and susceptibility through a genetic process going on in grapevine infected with the powdery mildew disease.

Infected grapevine expressed an upregulation of a gene that encodes a nitrate transporter, a protein that regulates the makes it possible for the protein to enter the plant cell.

Once the pathogen is attracted to this varietal of grapevine, it tricks grapevine into providing nutrients, allowing the mildew to grow and devastate the plant.

As leaves mature, they go through a transition where they’re no longer taking a lot of nutrients for themselves. Instead, they become “sources” and send nutrients to new “sink” leaves and tissues. The exchange enables plants to grow.

The powdery mildew pathogen, which requires a living host, tricks the grapevine into using its nutrient transfer against itself. Leaves turn into a “sink” for the pathogens, and nutrients that would have gone to new leaves, go instead, to the pathogen, Gassmann said.

“We think that what this fungus has to do is make this leaf a sink for nitrate so that nitrate goes to the pathogen instead of going to the rest of the plant,” Gassmann said.

Walter Gassmann, of the Bond Life Sciences Center at the University of Missouri was the lead investogator on the research. Much of his work has been on grapevine susceptibility to pathogens.

Walter Gassmann, of the Bond Life Sciences Center at the University of Missouri was the lead investogator on the research. Much of his work has been on grapevine susceptibility to pathogens. | Roger Meissen, Bond Life Sciences Center

According to a report by the USDA, powdery mildew can cause “major yield losses if infection occurs early in the crop cycle and conditions remain favorable for development.”

Powdery mildew appears as white to pale gray “fuzzy” blotches on the upper surfaces of leaves and thrives in “cool, humid and semiarid areas,” according to the report.

Gassmann said powdery mildew affects grapevine leaves, stems and berries and contributes to significant crop loss of the Vitas vinifera, which is cultivated for most commercial wine varietals.

“The leaves that are attacked lose their chlorophyll and they can’t produce much sugar,” Gassmann said. “Plus the grape berries get infected directly, so quality and yield are reduced in multiple ways.”

Pinpointing a cause

Solutions to problems start with finding the reason why something is happening, so Gassmann and his team looked at a list of genes activated by the pathogen to find transporters that allowed compounds like peptides, amino acids, and nitrate to pass.

Genes for nitrate transporters, Gassmann said, pointed to a cause for vulnerability to the mildew pathogen.

Over-fertilization of nitrate increases the severity of mildew in many crop plants, according to previous studies sited in Gassmann’s article in the journal of Plant Cell Physiology.

The testing system for isolating and analyzing the genes began with female frogs.

Gassmann used frog oocytes (unfertilized eggs), to verify the similar functions of nitrate transporters in Arabidopsis thaliana, a plant used as a baseline for comparison.

A nitrate transporter, he hypothesized, would increase the grapevine’s susceptibility to mildew.

“The genes that were upregulated in grapevine showed similarity to genes in Arabidopsis that are known to transport nitrate,” Gassmann said. “We felt the first thing we had to do was verify that what we have in grapevine actually does that.”

The eggs are very large relative to other testing systems and act as “an incubating system” for developing a protein. Gassmann and his team of researchers injected the oocyte with RNA, a messenger molecule that contains the information from a gene to produce a protein. The egg thinks it’s being fertilized and protein reproduces and is studied.

“The oocyte is like a machine to crank out protein,” Gassmann said. “We use that technique to establish what we have is actually a nitrate transporter.”

The system confirmed that the gene isolated from grapevine encodes a nitrate transporter.

“We contributed to the general knowledge of the nitrate transporter family,” Gassmann said. “It turned out to be the first member of one branch of nitrate transporters that, even in Arabidopsis haven’t been characterized before.”

The mounting knowledge of Vitis vinifera genes could make genetically modifying the strain to prevent the susceptibility easier.

“Resistance is determined sometimes by a single gene,” Gassmann said. “Until people are willing to have the conversation of genetic modification, the only way to save your grapevines is to be spraying a lot.”

Sharon Pike, Gassmann, other investigators from the MU Christopher S. Bond Life Sciences Center and post-doctoral student, Min Jung Kim from Daniel Schachtman’s lab at the Donald Danforth Plant Science Center in Saint Louis, Mo. contributed to the report.

The article was accepted November 2013 into the Plant Cell Physiology journal.

Choi honored for distinguished dissertation

Jeongmin Choi (left), Gary Stacey (center) and postdoc Kiwamu Tanaka recently discovered the first plant receptor for extracellular ATP.

Jeongmin Choi (left), Gary Stacey (center) and postdoc Kiwamu Tanaka recently discovered the first plant receptor for extracellular ATP. Choi received the 2014 Distinguished Dissertation Award for her part in this work.

A former Bond LSC graduate student is being recognized for a dissertation that stands out from the crowd.

Jeongmin Choi received the 2014 Distinguished Dissertation Award this month from MU’s Graduate Faculty Senate for her work identifying the first plant receptor for extracellular ATP. The journal Science published Choi’s “Identification of a plant receptor for extracellular ATP”  Jan. 17, 2014.

Choi completed her dissertation working as a member of Gary Stacey’s lab team. Stacey, a Bond LSC researcher, nominated her work for this award. This is the second year work completed in Bond LSC garnered this award after Lefteris Michailidis won in 2013 for work on the HIV drug EFdA.

Choi has since received her Ph.D. and now resides in Cambridge, England.

Read more about her work in Bond LSC team identifies first plant receptor for extracellular ATP published in January.

 

MU researchers find key gene in spinal locomotion, yield insight on paralysis

Samuel Waters and graduate researcher Desiré Buckley review stages of embryonic development.

Samuel Waters and graduate researcher Desiré Buckley review stages of embryonic development. — BLANKENBUEHLER

The difference between walking and being paralyzed could be as simple as turning a light switch on and off, a culmination of years of research shows.

Recently, University of Missouri Assistant Professor of biology Samuel T. Waters isolated a coding gene that he found has profound effects on locomotion and central nervous system development.

Waters’ work with gene expression in embryonic mouse tissue could shed light on paralysis and stroke and other disorders of the central nervous system, like Alzheimer’s disease.

Waters works extensively with two coding genes called “Gbx1” and “Gbx2”. These genes — exist in the body with approximately 20,000 other protein-coding genes — are essential for development in the central nervous system.

“To understand what’s going wrong, it’s critical that we know that’s right,” Waters said.

Coding genes essentially assign functions for the body. They tell your fingernail to grow a certain way, help develop motor control responsible for chewing and, as shown in Waters’ research, help your legs work with your spinal cord to facilitate movement.

Waters and his researchers, including graduate student Desiré Buckley, investigated the function of the Gbx1 by deactivating it in mouse embryos and observing their development over a 18.5-day gestation period — the time it takes a mouse to form.

The technology could eventually contribute to developing gene therapies for paralysis that happens at birth or from a direct result of blunt trauma, like a car accident.

“Understanding what allows us to walk normally and have motor control, allows us to have better insight for developing strategies for repairing neural circuits and therapies,” Waters said.

 

Technology for isolating genes and their functions

Waters studies embryonic mouse development. To understand certain gene functions, he inactivates different genes using a technology called “Cre-loxP.”

Genes can be isolated, then inactivated throughout embryonic tissue. Many of Waters’s studies inactivate genes to harness a better understanding of which genes are responsible for what.

“The relevance of it to the well-being of humans, is apparently relevant to development and more importantly to the development of the central nervous system,” Waters said. “Now it’s taking me to the point where we’re getting a bird’s eye view of what’s actually regulating our ability to have locomotive control.”

 

No Gbx1, no regular locomotion

Mice that Waters uses in his lab, “display a gross locomotive defect that specifically affects hind-limb gait,” according to their article published in Plos One, February, 2013.

In contrast to its family member Gbx2, when Gbx1 is inactivated, Waters concluded, the anterior hindbrain and cerebellum appear to develop normally. But neural circuit development in the spinal cord —- what allows us to walk normally —- is compromised, he said. According to an article published by

Waters, November 2013, in Methods in Molecular Biology, this occurs despite an increase in the expression level of its  family member, Gbx2, in the spinal cord.

A video recording from the research, which was funded by the National Science Foundation and start-up funds from MU, show the mouse with the Gbx1 held back, with an abnormal hind-limb-gait.

Mice with this inactivated gene were otherwise normal, Waters said.

“If they were sitting there without moving, you wouldn’t know anything was wrong with them,” Waters said.” They’re able to mate, eat and appear to function normally.”

Photographs taken during the research that show the hind limb gait defect in specimen with Gbx1 held back.

Photographs taken during the research that show the hind limb gait defect in specimen with Gbx1 held back.

No Gbx2, no jaw mobility

When Gbx2 function is impaired in the mouse, Waters observed that development of the anterior hindbrain, including the cerebellum, a region of the brain that plays an important role in motor control, didn’t form correctly.

The mice, as a result, cannot suckle, so they die at birth, Waters said.

“We’re getting a better insight into the requirements for suckling —   another motor function required for our survival,” Waters said.

The research has paved the way for investigating other coding genes and their responsibilities and roles in development, Waters said.

“We have a lot to do still,” Waters said. “So, why am I so excited about it? That’s part of the reason.”