Duke Research Blog

Following the people and events that make up the research community at Duke.

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Scientists Engineer Disease-Resistant Rice Without Sacrificing Yield

Researchers have developed a way to make rice more resistant to bacterial blight and other diseases without reducing yield. Photo by Max Pixel.

Researchers have successfully developed a novel method that allows for increased disease resistance in rice without decreasing yield. A team at Duke University, working in collaboration with scientists at Huazhong Agricultural University in China, describe the findings in a paper published May 17, 2017 in the journal Nature.

Rice is one of the most important staple crops, responsible for providing over one-fifth of the calories consumed by humans worldwide. Diseases caused by bacterial or fungal pathogens present a significant problem, and can result in the loss of 80 percent or more of a rice crop.

Decades of research into the plant immune response have identified components that can be used to engineer disease-resistant plants. However, their practical application to crops is limited due to the decreased yield associated with a constantly active defense response.

“Immunity is a double-edged sword, ” said study co-author Xinnian Dong, professor of biology at Duke and lead investigator of the study. “There is often a tradeoff between growth and defense because defense proteins are not only toxic to pathogens but also harmful to self when overexpressed,” Dong said. “This is a major challenge in engineering disease resistance for agricultural use because the ultimate goal is to protect the yield.”

Previous studies have focused on altering the coding sequence or upstream DNA sequence elements of a gene. These upstream DNA elements are known as promoters, and they act as switches that turn on or off a gene’s expression. This is the first step of a gene’s synthesis into its protein product, known as transcription.

By attaching a promoter that gives an “on” signal to a defense gene, a plant can be engineered to be highly resistant to pathogens, though at a cost to growth and yield. These costs can be partially alleviated by attaching the defense gene to a “pathogen specific” promoter that turns on in the presence of pathogen attack.

To further alleviate the negative effects of active defense, the Dong group sought to add an additional layer of control. They turned newly discovered sequence elements, called upstream open reading frames (uORFs), to help address this problem. These sequence elements act on the intermediate of a gene, or messenger (RNA, a molecule similar to DNA) to govern its “translation” into the final protein product. A recent study by the Dong lab in an accompanying paper in Nature has identified many of these elements that respond in a pathogen-inducible manner.

The Dong group hypothesized that adding this pathogen-inducible translational regulation would result in a tighter control of defense protein expression and minimize the lost yield associated with enhanced disease resistance.

To test this hypothesis, the researchers started with Arabidopsis, a flowering plant commonly used in laboratory research. They created a DNA sequence that contains both the transcriptional and translational elements (uORFs) and fused them upstream of the potent “immune activator” gene called snc1. This hybrid sequence was called a “transcriptional/translational cassette” and was inserted into Arabidopsis plants.

When plants have snc1 constitutively active, they are highly resistant to pathogens, but have severely stunted growth. Strikingly, plants with the transcriptional/translational cassette not only have increased resistance, but they also lacked growth defects and resembled healthy wild-type plants. These results show the benefits of adding translational control in engineering plants that have increased resistance without significant costs.

The Dong group then sought to apply these findings to engineer disease-resistant rice, as it is one of the world’s most important crops. They created transgenic rice lines containing the transcriptional/translational cassette driving expression of another potent “immune activator” gene called AtNPR1. This gene was chosen as it has been found to confer broad spectrum pathogen resistance in a wide variety of crop species, including rice, citrus, apple and wheat.

The dry yellowish leaves on these rice plants are a classic symptom of bacterial blight, a devastating disease that affects rice fields worldwide. Photo by Meng Yuan.

The transgenic rice lines containing the transcriptional/translational cassette were infected with bacterial/fungal pathogens that cause three major rice diseases — rice  blight, leaf streak, and fungal blast. These showed high resistance to all three pathogens, indicating broad spectrum resistance could be achieved. Importantly, when grown in the field, their yield — both in terms of grain quantity and quality per plant — was almost unaffected. These results indicate a great potential for agricultural applications.

This strategy is the first known use of adding translational control for the engineering of disease-resistant crops with minimal yield costs. It has many advantages, as it is broadly applicable to a variety of crop species against many pathogens. Since this strategy involves activating the plants’ endogenous defenses, it may also reduce the use of pesticides on crops and hence protect the environment.

Additionally, these findings may be broadly applicable to other systems as well. These upstream elements (uORFs) are widely present in organisms from yeast to humans, with nearly half of all human transcripts containing them. “The great potential in using these elements in controlling protein translation during specific biological processes has yet to be realized,” Dong said.

Corresponding author Xinnian Dong can be reached at xdong@duke.edu or (919) 613-8176.

CITATION:  “uORF-Mediated Translation Allows Engineered Plant Disease Resistance Without Fitness Costs,” Guoyong Xu, Meng Yuan,   Chaoren Ai, Lijing Liu, Edward Zhuang, Sargis Karapetyan, Shiping Wang and Xinnian Dong. Nature, May 17, 2017. DOI: 10.1038/nature22372

 

Guest post by Jonathan Motley

Where Some Ski, Others Do Science

For most people, Lost Trail is a ski spot located at 7,000 feet in the Rocky Mountains on the border of Idaho and Montana. Skiers and snowboarders descend down steep slopes, past forests and alpine meadows that get more than 25 feet of snow each year. But for a team of researchers led by Duke biology professor Thomas Mitchell-Olds, buried beneath the snow is a hidden population of native plants on the cusp of dividing into two new species.

Molly Rivera-Olds shovels snow at Lost Trail Pass.

Studying a spindly North American wildflower called Boechera stricta, Mitchell-Olds and colleagues suspected that a process called chromosomal inversion — in which part of a chromosome breaks off and reattaches itself upside down — plays a central role in speciation. To test the idea, they planted Boechera stricta seedlings in a mountaintop meadow near the Lost Trail resort.

To reach the meadow, the researchers carried thousands of seedlings up the mountain in specially constructed backpacks. They also lugged up nine empty garbage cans and filled them with snow to water the plants throughout the summer.

Once the seedlings matured, the researchers measured flowering time, seed production, and survival. They found that plants with the chromosomal inversion had a leg up on the steep slopes of the Rocky Mountains. Eventually, the researchers say, this can lead to plants with the inverted DNA splitting off and forming a new species.

The findings were published April 3, 2017 in the journal Nature Ecology & Evolution.

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CITATION:  “Young Inversion with Multiple Linked QTLs Under Selection in a Hybrid Zone,” Cheng-Ruei Lee, Baosheng Wang et al. Nature Ecology & Evolution, April 3, 2017. DOI:10.1038/s41559-017-0119.

Guest post by Molly Rivera-Olds

 

 

 

 

 

Data Geeks Go Head to Head

For North Carolina college students, “big data” is becoming a big deal. The proof: signups for DataFest, a 48-hour number-crunching competition held at Duke last weekend, set a record for the third time in a row this year.

DataFest 2017

More than 350 data geeks swarmed Bostock Library this weekend for a 48-hour number-crunching competition called DataFest. Photo by Loreanne Oh, Duke University.

Expected turnout was so high that event organizer and Duke statistics professor Mine Cetinkaya-Rundel was even required by state fire code to sign up for “crowd manager” safety training — her certificate of completion is still proudly displayed on her Twitter feed.

Nearly 350 students from 10 schools across North Carolina, California and elsewhere flocked to Duke’s West Campus from Friday, March 31 to Sunday, April 2 to compete in the annual event.

Teams of two to five students worked around the clock over the weekend to make sense of a single real-world data set. “It’s an incredible opportunity to apply the modeling and computing skills we learn in class to actual business problems,” said Duke junior Angie Shen, who participated in DataFest for the second time this year.

The surprise dataset was revealed Friday night. Just taming it into a form that could be analyzed was a challenge. Containing millions of data points from an online booking site, it was too large to open in Excel. “It was bigger than anything I’ve worked with before,” said NC State statistics major Michael Burton.

DataFest 2017

The mystery data set was revealed Friday night in Gross Hall. Photo by Loreanne Oh.

Because of its size, even simple procedures took a long time to run. “The dataset was so large that we actually spent the first half of the competition fixing our crushed software and did not arrive at any concrete finding until late afternoon on Saturday,” said Duke junior Tianlin Duan.

The organizers of DataFest don’t specify research questions in advance. Participants are given free rein to analyze the data however they choose.

“We were overwhelmed with the possibilities. There was so much data and so little time,” said NCSU psychology major Chandani Kumar.

“While for the most part data analysis was decided by our teachers before now, this time we had to make all of the decisions ourselves,” said Kumar’s teammate Aleksey Fayuk, a statistics major at NCSU.

As a result, these budding data scientists don’t just write code. They form theories, find patterns, test hunches. Before the weekend is over they also visualize their findings, make recommendations and communicate them to stakeholders.

This year’s participants came from more than 10 schools, including Duke, UNC, NC State and North Carolina A&T. Students from UC Davis and UC Berkeley also made the trek. Photo by Loreanne Oh.

“The most memorable moment was when we finally got our model to start generating predictions,” said Duke neuroscience and computer science double major Luke Farrell. “It was really exciting to see all of our work come together a few hours before the presentations were due.”

Consultants are available throughout the weekend to help with any questions participants might have. Recruiters from both start-ups and well-established companies were also on site for participants looking to network or share their resumes.

“Even as late as 11 p.m. on Saturday we were still able to find a professor from the Duke statistics department at the Edge to help us,” said Duke junior Yuqi Yun, whose team presented their results in a winning interactive visualization. “The organizers treat the event not merely as a contest but more of a learning experience for everyone.”

Caffeine was critical. “By 3 a.m. on Sunday morning, we ended initial analysis with what we had, hoped for the best, and went for a five-hour sleep in the library,” said NCSU’s Fayuk, whose team DataWolves went on to win best use of outside data.

By Sunday afternoon, every surface of The Edge in Bostock Library was littered with coffee cups, laptops, nacho crumbs, pizza boxes and candy wrappers. White boards were covered in scribbles from late-night brainstorming sessions.

“My team encouraged everyone to contribute ideas. I loved how everyone was treated as a valuable team member,” said Duke computer science and political science major Pim Chuaylua. She decided to sign up when a friend asked if she wanted to join their team. “I was hesitant at first because I’m the only non-stats major in the team, but I encouraged myself to get out of my comfort zone,” Chuaylua said.

“I learned so much from everyone since we all have different expertise and skills that we contributed to the discussion,” said Shen, whose teammates were majors in statistics, computer science and engineering. Students majoring in math, economics and biology were also well represented.

At the end, each team was allowed four minutes and at most three slides to present their findings to a panel of judges. Prizes were awarded in several categories, including “best insight,” “best visualization” and “best use of outside data.”

Duke is among more than 30 schools hosting similar events this year, coordinated by the American Statistical Association (ASA). The winning presentations and mystery data source will be posted on the DataFest website in May after all events are over.

The registration deadline for the next Duke DataFest will be March 2018.

DataFest 2017

Bleary-eyed contestants pose for a group photo at Duke DataFest 2017. Photo by Loreanne Oh.

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Post by Robin Smith

Closing the Funding Gap for Minority Scientists

DURHAM, N.C. — The barriers to minority students in science, technology, engineering and math (STEM) don’t go away once they’ve finished school and landed a job, studies show. But one nationwide initiative aims to level the playing field once they get there.

With support from a 3-year, $500,0000 grant from the National Science Foundation, assistant professors and postdoctoral fellows who come from underrepresented minorities are encouraged to apply by May 5 for a free grant writing workshop to be held June 22-24 in Washington, D.C..

It’s no secret that STEM has a diversity problem. In 2015, African-Americans and Latinos made up 29 percent of the U.S. workforce, but only 11 percent of scientists and engineers.

A study published in the journal Science in 2011 revealed that minority scientists also were less likely to win grants from the National Institutes of Health, the largest source of research funding to universities.

Based on an analysis of 83,000 grant applications from 2000 to 2006, the study authors found that applications from black researchers were 13 percent less likely to succeed than applications from their white peers. Applications from Asian and Hispanic scientists were 5 and 3 percent less likely to be awarded, respectively.

Even when the study authors made sure they were comparing applicants with similar educational backgrounds, training, employers and publication records, the funding gap persisted — particularly for African-Americans.

Competition for federal research dollars is already tough. But white scientists won 29 percent of the time, and black scientists succeeded only 16 percent of the time.

Pennsylvania State University chemistry professor Squire Booker is co-principal investigator of a $500,000 initiative funded by the National Science Foundation to help underrepresented minority scientists write winning research grants.

“That report sent a shock wave through the scientific community,” said Squire Booker, a Howard Hughes Medical Institute investigator and chemistry professor at Pennsylvania State University. Speaking last week in the Nanaline H. Duke building on Duke’s Research Drive, Booker outlined a mentoring initiative that aims to close the gap.

In 2013, Booker and colleagues on the Minority Affairs Committee of the American Society for Biochemistry and Molecular Biology decided to host a workshop to demystify the grant application process and help minority scientists write winning grants.

Grant success is key to making it in academia. Even at universities that don’t make funding a formal requirement for tenure and promotion, research is expensive. Outside funding is often required to keep a lab going, and research productivity — generating data and publishing results — is critical.

To insure underrepresented minorities have every chance to compete for increasingly tight federal research dollars, Booker and colleagues developed the Interactive Mentoring Activities for Grantsmanship Enhancement program, known as IMAGE. Program officers from NIH and NSF offer tips on navigating the funding process, crafting a successful proposal, decoding reviews and revising and resubmitting. The organizers also stage a mock review panel, and participants receive real-time, constructive feedback on potential research proposals.

Participants include researchers in biology, biophysics, biochemistry and molecular biology. More than half of the program’s 130 alumni have been awarded NSF or NIH grants since the workshop series started in 2013.

Booker anticipates this year’s program will include more postdoctoral fellows. “Now we’re trying to expand the program to intervene at an earlier stage,” Booker said.

To apply for the 2017 workshop visit http://www.asbmb.org/grantwriting/.  The application deadline is May 5.

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Post by Robin Smith

Seeing Nano

Take pictures at more than 300,000 times magnification with electron microscopes at Duke

Sewer gnat head

An image of a sewer gnat’s head taken through a scanning electron microscope. Courtesy of Fred Nijhout.

The sewer gnat is a common nuisance around kitchen and bathroom drains that’s no bigger than a pea. But magnified thousands of times, its compound eyes and bushy antennae resemble a first place winner in a Movember mustache contest.

Sewer gnats’ larger cousins, horseflies are known for their painful bite. Zoom in and it’s easy to see how they hold onto their furry livestock prey:  the tiny hooked hairs on their feet look like Velcro.

Students in professor Fred Nijhout’s entomology class photograph these and other specimens at more than 300,000 times magnification at Duke’s Shared Material & Instrumentation Facility (SMIF).

There the insects are dried, coated in gold and palladium, and then bombarded with a beam of electrons from a scanning electron microscope, which can resolve structures tens of thousands of times smaller than the width of a human hair.

From a ladybug’s leg to a weevil’s suit of armor, the bristly, bumpy, pitted surfaces of insects are surprisingly beautiful when viewed up close.

“The students have come to treat travels across the surface of an insect as the exploration of a different planet,” Nijhout said.

Horsefly foot

The foot of a horsefly is equipped with menacing claws and Velcro-like hairs that help them hang onto fur. Photo by Valerie Tornini.

Weevil

The hard outer skeleton of a weevil looks smooth and shiny from afar, but up close it’s covered with scales and bristles. Courtesy of Fred Nijhout.

fruit fly wing

Magnified 500 times, the rippled edges of this fruit fly wing are the result of changes in the insect’s genetic code. Courtesy of Eric Spana.

You, too, can gaze at alien worlds too small to see with the naked eye. Students and instructors across campus can use the SMIF’s high-powered microscopes and other state of the art research equipment at no charge with support from the Class-Based Explorations Program.

Biologist Eric Spana’s experimental genetics class uses the microscopes to study fruit flies that carry genetic mutations that alter the shape of their wings.

Students in professor Hadley Cocks’ mechanical engineering 415L class take lessons from objects that break. A scanning electron micrograph of a cracked cymbal once used by the Duke pep band reveals grooves and ridges consistent with the wear and tear from repeated banging.

cracked cymbal

Magnified 3000 times, the surface of this broken cymbal once used by the Duke Pep Band reveals signs of fatigue cracking. Courtesy of Hadley Cocks.

These students are among more than 200 undergraduates in eight classes who benefitted from the program last year, thanks to a grant from the Donald Alstadt Foundation.

You don’t have to be a scientist, either. Historians and art conservators have used scanning electron microscopes to study the surfaces of Bronze Age pottery, the composition of ancient paints and even dust from Egyptian mummies and the Shroud of Turin.

Instructors and undergraduates are invited to find out how they could use the microscopes and other nanotech equipement in the SMIF in their teaching and research. Queries should be directed to Dr. Mark Walters, Director of SMIF, via email at mark.walters@duke.edu.

Located on Duke’s West Campus in the Fitzpatrick Building, the SMIF is a shared use facility available to Duke researchers and educators as well as external users from other universities, government laboratories or industry through a partnership called the Research Triangle Nanotechnology Network. For more info visit http://smif.pratt.duke.edu/.

Scanning electron microscope

This scanning electron microscope could easily be mistaken for equipment from a dentist’s office.

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Post by Robin Smith

How to Get a Lemur to Notice You

Duke evolutionary anthropology professor Brian Hare studies what goes on in the minds of animals.

Duke evolutionary anthropology professor Brian Hare studies what goes on in the minds of animals.

Duke professor Brian Hare remembers his first flopped experiment. While an undergraduate at Emory in the late 1990s, he spent a week at the Duke Lemur Center waving bananas at lemurs. He was trying to see if they, like other primates, possess an important social skill. If a lemur spots a piece of food, or a predator, can other lemurs follow his gaze to spot it too?

First he needed the lemurs to notice him. If he could get one lemur to look at him, he could figure out if other lemurs then turn around and look too. In similar experiments with monkeys and chimps, oranges had done the trick.

“But I couldn’t get their attention,” Hare said. “It failed miserably.”

Hare was among more than 200 people from 25 states and multiple countries who converged in Durham this week for the 50th anniversary celebration of the Duke Lemur Center, Sept. 21-23, 2016.

Humans look to subtle movements in faces and eyes for clues to what others are thinking, Hare told a crowd assembled at a two-day research symposium held in conjunction with the event.

If someone quickly glances down at your name tag, for example, you can guess just from that eye movement that they can’t recall your name.

We develop this skill as infants. Most kids start to follow the gaze of others by the age of two. A lack of interest in gaze-following is considered an early sign of autism.

Arizona State University graduate student Joel Bray got hooked on lemurs while working as an undergraduate research assistant in the Hare lab.

Arizona State University graduate student Joel Bray got hooked on lemurs while working as an undergraduate research assistant in the Hare lab.

“Gaze-following suggests that kids are starting to think about the thoughts of others,” Hare said. “And using where others look to try to understand what they want or what they know.”

In 1998 Hare and researchers Michael Tomasello and Josep Call published a study showing that chimpanzees and multiple species of monkeys are able to look where others are looking. But at the time not much was known about cognition in lemurs.

“When you study dogs you just say, ‘sit, stay,’ and they’re happy to play along,” Hare said. Working at the Duke Lemur Center, eventually his students discovered the secret to making these tree-dwelling animals feel at home: “Lemurs like to be off the ground,” Hare said. “We figured out that if we just let them solve problems on tables, they’re happy to participate.”

Studies have since shown that multiple lemur species are able to follow the gaze of other lemurs. “Lemurs have gone from ignored to adored in cognitive research,” Hare said.

 

Ring-tailed lemurs are among several species of lemurs known to follow the gaze of other lemurs. The ability to look where others are looking is considered a key step towards understanding what others see, know, or might do. Photo by David Haring, Duke Lemur Center.

Ring-tailed lemurs are among several lemur species known to follow the gaze of other lemurs. The ability to look where others are looking is considered a key step towards understanding what others see, know, or might do. Photo by David Haring, Duke Lemur Center.

Robin SmithPost by Robin A. Smith

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