Nanosensor can alert a smartphone when plants are stressed

Carbon nanotubes embedded in leaves detect chemical signals that are produced when a plant is damaged.

Oli Scarff/Getty Images
MIT engineers have developed a way to closely track how plants respond to stresses such as injury, infection, and light damage, using sensors made of carbon nanotubes.

These sensors can be embedded in plant leaves, where they report on hydrogen peroxide signaling waves.

Plants use hydrogen peroxide to communicate within their leaves, sending out a distress signal that stimulates leaf cells to produce compounds that will help them repair damage or fend off predators such as insects. The new sensors can use these hydrogen peroxide signals to distinguish between different types of stress, as well as between different species of plants.

"Plants have a very sophisticated form of internal communication, which we can now observe for the first time. That means that in real-time, we can see a living plant's response, communicating the specific type of stress that it's experiencing," says Michael Strano, the Carbon P. Dubbs Professor of Chemical Engineering at MIT.

This kind of sensor could be used to study how plants respond to different types of stress, potentially helping agricultural scientists develop new strategies to improve crop yields. The researchers demonstrated their approach in eight different plant species, including spinach, strawberry plants, and arugula, and they believe it could work in many more.

Strano is the senior author of the study, which appears today in Nature Plants. MIT graduate student Tedrick Thomas Salim Lew is the lead author of the paper.

Embedded sensors

Over the past several years, Strano's lab has been exploring the potential for engineering "nanobionic plants" — plants that incorporate nanomaterials that give the plants new functions, such as emitting light or detecting water shortages. In the new study, he set out to incorporate sensors that would report back on the plants' health status.

Strano had previously developed carbon nanotube sensors that can detect various molecules, including hydrogen peroxide. About three years ago, Lew began working on trying to incorporate these sensors into plant leaves. Studies in Arabidopsis thaliana, often used for molecular studies of plants, had suggested that plants might use hydrogen peroxide as a signaling molecule, but its exact role was unclear.

Lew used a method called lipid exchange envelope penetration (LEEP) to incorporate the sensors into plant leaves. LEEP, which Strano's lab developed several years ago, allows for the design of nanoparticles that can penetrate plant cell membranes. As Lew was working on embedding the carbon nanotube sensors, he made a serendipitous discovery.

"I was training myself to get familiarized with the technique, and in the process of the training I accidentally inflicted a wound on the plant. Then I saw this evolution of the hydrogen peroxide signal," he says.

He saw that after a leaf was injured, hydrogen peroxide was released from the wound site and generated a wave that spread along the leaf, similar to the way that neurons transmit electrical impulses in our brains. As a plant cell releases hydrogen peroxide, it triggers calcium release within adjacent cells, which stimulates those cells to release more hydrogen peroxide.

"Like dominos successively falling, this makes a wave that can propagate much further than a hydrogen peroxide puff alone would," Strano says. "The wave itself is powered by the cells that receive and propagate it."

This flood of hydrogen peroxide stimulates plant cells to produce molecules called secondary metabolites, such as flavonoids or carotenoids, which help them to repair the damage. Some plants also produce other secondary metabolites that can be secreted to fend off predators. These metabolites are often the source of the food flavors that we desire in our edible plants, and they are only produced under stress.

A key advantage of the new sensing technique is that it can be used in many different plant species. Traditionally, plant biologists have done much of their molecular biology research in certain plants that are amenable to genetic manipulation, including Arabidopsis thaliana and tobacco plants. However, the new MIT approach is applicable to potentially any plant.

"In this study, we were able to quickly compare eight plant species, and you would not be able to do that with the old tools," Strano says.

The researchers tested strawberry plants, spinach, arugula, lettuce, watercress, and sorrel, and found that different species appear to produce different waveforms — the distinctive shape produced by mapping the concentration of hydrogen peroxide over time. They hypothesize that each plant's response is related to its ability to counteract the damage. Each species also appears to respond differently to different types of stress, including mechanical injury, infection, and heat or light damage.

"This waveform holds a lot of information for each species, and even more exciting is that the type of stress on a given plant is encoded in this waveform," Strano says. "You can look at the real time response that a plant experiences in almost any new environment."

Stress response

The near-infrared fluorescence produced by the sensors can be imaged using a small infrared camera connected to a Raspberry Pi, a $35 credit-card-sized computer similar to the computer inside a smartphone. "Very inexpensive instrumentation can be used to capture the signal," Strano says.

Applications for this technology include screening different species of plants for their ability to resist mechanical damage, light, heat, and other forms of stress, Strano says. It could also be used to study how different species respond to pathogens, such as the bacteria that cause citrus greening and the fungus that causes coffee rust.

"One of the things I'm interested in doing is understanding why some types of plants exhibit certain immunity to these pathogens and others don't," he says.

Strano and his colleagues in the Disruptive and Sustainable Technology for Agricultural Precision interdisciplinary research group at the Singapore-MIT Alliance for Research and Technology (SMART), MIT's research enterprise in Singapore, are also interested in studying is how plants respond to different growing conditions in urban farms.

One problem they hope to address is shade avoidance, which is seen in many species of plants when they are grown at high density. Such plants turn on a stress response that diverts their resources into growing taller, instead of putting energy into producing crops. This lowers the overall crop yield, so agricultural researchers are interested in engineering plants so that don't turn on that response.

"Our sensor allows us to intercept that stress signal and to understand exactly the conditions and the mechanism that are happening upstream and downstream in the plant that gives rise to the shade avoidance," Strano says.

The research was funded by the National Research Foundation of Singapore, the Singapore Agency for Science, Technology, and Research (A*STAR), and the U.S. Department of Energy Computational Science Graduate Fellowship Program.

Reprinted with permission of MIT News. Read the original article.

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New research establishes an unexpected connection.

Reactive oxygen species (ROS) accumulate in the gut of sleep-deprived fruit flies, one (left), seven (center) and ten (right) days without sleep.

Image source: Vaccaro et al, 2020/Harvard Medical School
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We don't have to tell you what it feels like when you don't get enough sleep. A night or two of that can be miserable; long-term sleeplessness is out-and-out debilitating. Though we know from personal experience that we need sleep — our cognitive, metabolic, cardiovascular, and immune functioning depend on it — a lack of it does more than just make you feel like you want to die. It can actually kill you, according to study of rats published in 1989. But why?

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The study, from researchers at Harvard Medical School (HMS), is published in the journal Cell.

An unexpected culprit

The new research examines the mechanisms at play in sleep-deprived fruit flies and in mice — long-term sleep-deprivation experiments with humans are considered ethically iffy.

What the scientists found is that death from sleep deprivation is always preceded by a buildup of Reactive Oxygen Species (ROS) in the gut. These are not, as their name implies, living organisms. ROS are reactive molecules that are part of the immune system's response to invading microbes, and recent research suggests they're paradoxically key players in normal cell signal transduction and cell cycling as well. However, having an excess of ROS leads to oxidative stress, which is linked to "macromolecular damage and is implicated in various disease states such as atherosclerosis, diabetes, cancer, neurodegeneration, and aging." To prevent this, cellular defenses typically maintain a balance between ROS production and removal.

"We took an unbiased approach and searched throughout the body for indicators of damage from sleep deprivation," says senior study author Dragana Rogulja, admitting, "We were surprised to find it was the gut that plays a key role in causing death." The accumulation occurred in both sleep-deprived fruit flies and mice.

"Even more surprising," Rogulja recalls, "we found that premature death could be prevented. Each morning, we would all gather around to look at the flies, with disbelief to be honest. What we saw is that every time we could neutralize ROS in the gut, we could rescue the flies." Fruit flies given any of 11 antioxidant compounds — including melatonin, lipoic acid and NAD — that neutralize ROS buildups remained active and lived a normal length of time in spite of sleep deprivation. (The researchers note that these antioxidants did not extend the lifespans of non-sleep deprived control subjects.)

fly with thought bubble that says "What? I'm awake!"

Image source: Tomasz Klejdysz/Shutterstock/Big Think

The experiments

The study's tests were managed by co-first authors Alexandra Vaccaro and Yosef Kaplan Dor, both research fellows at HMS.

You may wonder how you compel a fruit fly to sleep, or for that matter, how you keep one awake. The researchers ascertained that fruit flies doze off in response to being shaken, and thus were the control subjects induced to snooze in their individual, warmed tubes. Each subject occupied its own 29 °C (84F) tube.

For their sleepless cohort, fruit flies were genetically manipulated to express a heat-sensitive protein in specific neurons. These neurons are known to suppress sleep, and did so — the fruit flies' activity levels, or lack thereof, were tracked using infrared beams.

Starting at Day 10 of sleep deprivation, fruit flies began dying, with all of them dead by Day 20. Control flies lived up to 40 days.

The scientists sought out markers that would indicate cell damage in their sleepless subjects. They saw no difference in brain tissue and elsewhere between the well-rested and sleep-deprived fruit flies, with the exception of one fruit fly.

However, in the guts of sleep-deprived fruit flies was a massive accumulation of ROS, which peaked around Day 10. Says Vaccaro, "We found that sleep-deprived flies were dying at the same pace, every time, and when we looked at markers of cell damage and death, the one tissue that really stood out was the gut." She adds, "I remember when we did the first experiment, you could immediately tell under the microscope that there was a striking difference. That almost never happens in lab research."

The experiments were repeated with mice who were gently kept awake for five days. Again, ROS built up over time in their small and large intestines but nowhere else.

As noted above, the administering of antioxidants alleviated the effect of the ROS buildup. In addition, flies that were modified to overproduce gut antioxidant enzymes were found to be immune to the damaging effects of sleep deprivation.

The research leaves some important questions unanswered. Says Kaplan Dor, "We still don't know why sleep loss causes ROS accumulation in the gut, and why this is lethal." He hypothesizes, "Sleep deprivation could directly affect the gut, but the trigger may also originate in the brain. Similarly, death could be due to damage in the gut or because high levels of ROS have systemic effects, or some combination of these."

The HMS researchers are now investigating the chemical pathways by which sleep-deprivation triggers the ROS buildup, and the means by which the ROS wreak cell havoc.

"We need to understand the biology of how sleep deprivation damages the body so that we can find ways to prevent this harm," says Rogulja.

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