Tag Archives: Arabidopsis thaliana

The devil’s (i.e., luciferase) in the bioluminescent plant

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The American Chemical Society (ACS) and the Massachusetts Institute of Technology (MIT) have both issued news releases about the latest in bioluminescence.The researchers tested their work on watercress, a vegetable that was viewed in almost sacred terms in my family; it was not easily available in Vancouver (Canada) when I was child.

My father would hunt down fresh watercress by checking out the Chinese grocery stores. He could spot the fresh stuff from across the street while driving at 30 miles or more per hour. Spotting it entailed an immediate hunt for parking (my father hated to pay so we might have go around the block a few times or more) and a dash out of the car to ensure that he got his watercress before anyone else spotted it. These days it’s much more easily available and, thankfully, my father has passed on so he won’t have to think about glowing watercress.

Getting back to bioluninescent vegetable research, the American Chemical Society’s Dec. 13, 2017 news release on EurekAlert (and as a Dec. 13, 2017 news item on ScienceDaily) makes the announcement,

The 2009 film “Avatar” created a lush imaginary world, illuminated by magical, glowing plants. Now researchers are starting to bring this spellbinding vision to life to help reduce our dependence on artificial lighting. They report in ACS’ journal Nano Letters a way to infuse plants with the luminescence of fireflies.

Nature has produced many bioluminescent organisms, however, plants are not among them. Most attempts so far to create glowing greenery — decorative tobacco plants in particular — have relied on introducing the genes of luminescent bacteria or fireflies through genetic engineering. But getting all the right components to the right locations within the plants has been a challenge. To gain better control over where light-generating ingredients end up, Michael S. Strano and colleagues recently created nanoparticles that travel to specific destinations within plants. Building on this work, the researchers wanted to take the next step and develop a “nanobionic,” glowing plant.

The team infused watercress and other plants with three different nanoparticles in a pressurized bath. The nanoparticles were loaded with light-emitting luciferin; luciferase, which modifies luciferin and makes it glow; and coenzyme A, which boosts luciferase activity. Using size and surface charge to control where the sets of nanoparticles could go within the plant tissues, the researchers could optimize how much light was emitted. Their watercress was half as bright as a commercial 1 microwatt LED and 100,000 times brighter than genetically engineered tobacco plants. Also, the plant could be turned off by adding a compound that blocks luciferase from activating luciferin’s glow.

Here’s a video from MIT detailing their research,

A December 13, 2017 MIT news release (also on EurekAlert) casts more light on the topic (I couldn’t resist the word play),

Imagine that instead of switching on a lamp when it gets dark, you could read by the light of a glowing plant on your desk.

MIT engineers have taken a critical first step toward making that vision a reality. By embedding specialized nanoparticles into the leaves of a watercress plant, they induced the plants to give off dim light for nearly four hours. They believe that, with further optimization, such plants will one day be bright enough to illuminate a workspace.

“The vision is to make a plant that will function as a desk lamp — a lamp that you don’t have to plug in. The light is ultimately powered by the energy metabolism of the plant itself,” says Michael Strano, the Carbon P. Dubbs Professor of Chemical Engineering at MIT and the senior author of the study

This technology could also be used to provide low-intensity indoor lighting, or to transform trees into self-powered streetlights, the researchers say.

MIT postdoc Seon-Yeong Kwak is the lead author of the study, which appears in the journal Nano Letters.

Nanobionic plants

Plant nanobionics, a new research area pioneered by Strano’s lab, aims to give plants novel features by embedding them with different types of nanoparticles. The group’s goal is to engineer plants to take over many of the functions now performed by electrical devices. The researchers have previously designed plants that can detect explosives and communicate that information to a smartphone, as well as plants that can monitor drought conditions.

Lighting, which accounts for about 20 percent of worldwide energy consumption, seemed like a logical next target. “Plants can self-repair, they have their own energy, and they are already adapted to the outdoor environment,” Strano says. “We think this is an idea whose time has come. It’s a perfect problem for plant nanobionics.”

To create their glowing plants, the MIT team turned to luciferase, the enzyme that gives fireflies their glow. Luciferase acts on a molecule called luciferin, causing it to emit light. Another molecule called co-enzyme A helps the process along by removing a reaction byproduct that can inhibit luciferase activity.

The MIT team packaged each of these three components into a different type of nanoparticle carrier. The nanoparticles, which are all made of materials that the U.S. Food and Drug Administration classifies as “generally regarded as safe,” help each component get to the right part of the plant. They also prevent the components from reaching concentrations that could be toxic to the plants.

The researchers used silica nanoparticles about 10 nanometers in diameter to carry luciferase, and they used slightly larger particles of the polymers PLGA and chitosan to carry luciferin and coenzyme A, respectively. To get the particles into plant leaves, the researchers first suspended the particles in a solution. Plants were immersed in the solution and then exposed to high pressure, allowing the particles to enter the leaves through tiny pores called stomata.

Particles releasing luciferin and coenzyme A were designed to accumulate in the extracellular space of the mesophyll, an inner layer of the leaf, while the smaller particles carrying luciferase enter the cells that make up the mesophyll. The PLGA particles gradually release luciferin, which then enters the plant cells, where luciferase performs the chemical reaction that makes luciferin glow.

The researchers’ early efforts at the start of the project yielded plants that could glow for about 45 minutes, which they have since improved to 3.5 hours. The light generated by one 10-centimeter watercress seedling is currently about one-thousandth of the amount needed to read by, but the researchers believe they can boost the light emitted, as well as the duration of light, by further optimizing the concentration and release rates of the components.

Plant transformation

Previous efforts to create light-emitting plants have relied on genetically engineering plants to express the gene for luciferase, but this is a laborious process that yields extremely dim light. Those studies were performed on tobacco plants and Arabidopsis thaliana, which are commonly used for plant genetic studies. However, the method developed by Strano’s lab could be used on any type of plant. So far, they have demonstrated it with arugula, kale, and spinach, in addition to watercress.

For future versions of this technology, the researchers hope to develop a way to paint or spray the nanoparticles onto plant leaves, which could make it possible to transform trees and other large plants into light sources.

“Our target is to perform one treatment when the plant is a seedling or a mature plant, and have it last for the lifetime of the plant,” Strano says. “Our work very seriously opens up the doorway to streetlamps that are nothing but treated trees, and to indirect lighting around homes.”

The researchers have also demonstrated that they can turn the light off by adding nanoparticles carrying a luciferase inhibitor. This could enable them to eventually create plants that shut off their light emission in response to environmental conditions such as sunlight, the researchers say.

Here’s a link to and a citation for the paper,

A Nanobionic Light-Emitting Plant by Seon-Yeong Kwak, Juan Pablo Giraldo, Min Hao Wong, Volodymyr B. Koman, Tedrick Thomas Salim Lew, Jon Ell, Mark C. Weidman, Rosalie M. Sinclair, Markita P. Landry, William A. Tisdale, and Michael S. Strano. Nano Lett., 2017, 17 (12), pp 7951–7961 DOI: 10.1021/acs.nanolett.7b04369 Publication Date (Web): November 17, 2017

Copyright © 2017 American Chemical Society

This paper is behind a paywall.

Inside-out plants show researchers how cellulose forms

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Strictly speaking this story of tricking cellulose into growing on the surface rather than the interior of a cell is not a nanotechnology topic but I imagine that the folks who research nanocellulose materials will find this work of great interest. An Oct. 8, 2015 news item on ScienceDaily describes the research,

Researchers have been able to watch the interior cells of a plant synthesize cellulose for the first time by tricking the cells into growing on the plant’s surface.

“The bulk of the world’s cellulose is produced within the thickened secondary cell walls of tissues hidden inside the plant body,” says University of British Columbia Botany PhD candidate Yoichiro Watanabe, lead author of the paper published this week in Science.

“So we’ve never been able to image the cells in high resolution as they produce this all-important biological material inside living plants.”

An Oct. 8, 2015 University of British Columbia (UBC) news release on EurekAlert, which originated the news item, explains the interest in cellulose,

Cellulose, the structural component of cell walls that enables plants to stay upright, is the most abundant biopolymer on earth. It’s a critical resource for pulp and paper, textiles, building materials, and renewable biofuels.

“In order to be structurally sound, plants have to lay down their secondary cell walls very quickly once the plant has stopped growing, like a layer of concrete with rebar,” says UBC botanist Lacey Samuels, one of the senior authors on the paper.

“Based on our study, it appears plant cells need both a high density of the enzymes that create cellulose, and their rapid movement across the cell surface, to make this happen so quickly.”

This work, the culmination of years of research by four UBC graduate students supervised by UBC Forestry researcher Shawn Mansfield and Samuels, was facilitated by a collaboration with the Nara Institute of Technology in Japan to create the special plant lines, and researchers at the Carnegie Institution for Science at Stanford University to conduct the live cell imaging.

“This is a major step forward in our understanding of how plants synthesize their walls, specifically cellulose,” says Mansfield. “It could have significant implications for the way plants are bred or selected for improved or altered cellulose ultrastructural traits – which could impact industries ranging from cellulose nanocrystals to toiletries to structural building products.”

The researchers used a modified line of Arabidopsis thaliana, a small flowering plant related to cabbage and mustard, to conduct the experiment. The resulting plants look exactly like their non-modified parents, until they are triggered to make secondary cell walls on their exterior.

One of the other partners in this research, Stanford University’s Carnegie Institution of Science published an Oct. 8, 2015 news release on EurekAlert focusing on other aspects of the research (Note: Some of this is repetitive),

Now scientists, including Carnegie’s David Ehrhardt and Heather Cartwright, have exploited a new way to watch the trafficking of the proteins that make cellulose in the formation cell walls in real time. They found that organization of this trafficking by structural proteins called microtubules, combined with the high density and rapid rate of these cellulose producing enzymes explains how thick and high strength secondary walls are built. This basic knowledge helps us understand plants can stand upright, which was essential for the move of plants from the sea to the land, and may useful for engineering plants with improved mechanical properties for to increase yields or to produce novel bio-materials. The research is published in Science.

The live-cell imaging was conducted at Carnegie with colleagues from the University of British Columbia (UBC) using customized high-end instrumentation. For the first time, it directly tracked cellulose production to observe how xylem cells, cells that transport water and some nutrients, make cellulose for their secondary cell walls. Strong walls are based on a high density of enzymes that catalyze the synthesis of cellulose (called cellulose synthase enzymes) and their rapid movement across the xylem cell surface.

Watching xylem cells lay down cellulose in real time has not been possible before, because the vascular tissues of plants are hidden inside the plant body. Lead author Yoichiro Watanabe of UBC applied a system developed by colleagues at the Nara Institute of Science and Technology to trick plants into making xylem cells on their surface. The researchers fluorescently tagged a cellulose synthase enzyme of the experimental plant Arabidopsis to track the activity using high-end microscopes.

“For me, one of the most exciting aspects of this study was being able to observe how the microtubule cytoskeleton was actively directing the synthesis of the new cell walls at the level of individual enzymes. We can guess how a complex cellular process works from static snapshots, which is what we usually have had to work from in biology, but you can’t really understand the process until you can see it in action. ” remarked Carnegie’s David Ehrhardt.

Here’s a link to and a citation for the paper,

Visualization of cellulose synthases in Arabidopsis secondary cell walls by Y. Watanabe, M. J. Meents, L. M. McDonnell, S. Barkwill, A. Sampathkumar, H. N. Cartwright, T. Demura, D. W. Ehrhardt, A.L. Samuels, & S. D. Mansfield. Science 9 October 2015: Vol. 350 no. 6257 pp. 198-203 DOI: 10.1126/science.aac7446

This paper is behind a paywall.

With all of this talk of visualization, it’s only right that the researchers have made an image from their work available,

Caption: An image of artificially-produced cellulose in cells on the surface of a modified Arabidopsis thaliana plant. Credit: University of British Columbia.

Caption: An image of artificially-produced cellulose in cells on the surface of a modified Arabidopsis thaliana plant. Credit: University of British Columbia.