First step to reduce plant need for nitrogen fertilizer uncovered

Sep. 27, 2013 — Nitrogen fertilizer costs U.S. farmers approximately $8 billion each year, and excess fertilizer can find its way into rivers and streams, damaging the delicate water systems. Now, a discovery by a team of University of Missouri researchers could be the first step toward helping crops use less nitrogen, benefitting both farmers' bottom lines and the environment. The journal Science published the research this month.

Gary Stacey, an investigator in the MU Bond Life Sciences Center and professor of plant sciences in the College of Agriculture, Food and Natural Resources, found that crops, such as corn, are "confused" when confronted with an invasive, but beneficial, bacteria known as rhizobia bacteria. When the bacteria interact correctly with a crop, the bacteria receive some food from the plant and, simultaneously, produce nitrogen that most plants need. In his study, Stacey found that many other crops recognize the bacteria, but do not attempt to interact closely with them.

"The problem is that corn, tomatoes and other crops have a different response and don't support an intimate interaction with the rhizobia, thus making farmers apply larger amounts of nitrogen than might otherwise be necessary," Stacey said. "Scientists have known about this beneficial relationship since 1888, but it only exists in legume crops, like soybeans and alfalfa. We're working to transfer this trait to other plants like corn, wheat or rice, which we believe is possible since these other plants recognize the bacteria. It's a good first step."

When legumes like soybeans sense a signal from the bacteria, they create nodules where the bacteria gather and produce atmospheric nitrogen that the plants can then use to stimulate their growth. This reaction doesn't happen in other plants.

"There's this back and forth battle between a plant and a pathogen," said Yan Liang, a co-author of the study and post-doctoral fellow at MU. "Rhizobia eventually developed a chemical to inhibit the defense response in legumes and make those plants recognize it as a friend. Meanwhile, corn, tomatoes and other crops are still trying to defend themselves against this bacteria."

In the study, Stacey and Liang treated corn, soybeans, tomatoes and other plants to see how they responded when exposed to the chemical signal from the rhizobia bacteria. They found that the plants did receive the signal and, like legumes, inhibited the normal plant immune system. However, soybeans, corn and these other plants don't complete the extra step of forming nodules to allow the bacteria to thrive.

"The important finding was that these other plants didn't just ignore the rhizobia bacteria," Stacey said. "They recognized it, but just activated a different mechanism. Our next step is to determine how we can make the plants understand that this is a beneficial relationship and get them to activate a different mechanism that will produce the nodules that attract the bacteria instead of trying to fight them."

The study was funded by a grant from the U.S. Department of Energy.??

View the original article here

Plants in space: A novel method for fixing plant tissue samples maximizes time, resources, and data

Sep. 11, 2013 — At work on the International Space Station, researchers studying plant and cell growth in space encountered a challenge. Imaging revealed interesting spaceflight-associated root morphologies. They needed to fix the tissues for further study back on Earth, but conventional fixation methods require separate fixatives depending on whether the sample is intended for molecular or morphological study. If the scientists wanted to study how spaceflight affected patterns of gene expression central to morphological patterns of cell growth, they needed a fixation method that would allow them to study both perspectives.

Most scientists at work in the laboratory rely on protocols that have been developed without the need for restrictions on the amount of space, time, or reagents they use. For scientists conducting experiments in spaceflight, time and resources are strictly regulated and limited, and researchers must know in advance which protocols will maximize the usefulness of the data they collect.

University of Florida professors Anna-Lisa Paul and Robert Ferl and colleagues are collaborating with the National Aeronautics and Space Administration (NASA) to understand plant growth and development in spaceflight. Along with lead author and graduate student Eric Schultz, they have developed a single fixation protocol for use in space that allows plant material to be used for multiple experimental applications. Their new protocol for sample preparation was tested on Arabidopsis tissues harvested on the International Space Station and is described in the August issue of Applications in Plant Sciences.

Because of limitations in astronaut crew time and orbital resources, previous spaceflight fixation protocols were designated as either molecular or morphological, due to the separate fixatives required for each application. Tissues for morphologic study were fixed in 3% glutaraldehyde (or a similar solution), and tissues for molecular study were fixed in the tissue storage reagent RNAlater. RNAlater has not commonly been used as a morphologic fixative, as it can produce unclear images with high background staining.

The new method developed by Schultz et al. puts RNAlater-fixed samples through a desalination process to return them to a fresh-like state, and then uses low-temperature scanning electron microscopy (cryo-SEM) to preserve tissues for imaging. Because few laboratories have access to the necessary equipment for cryo-SEM, the authors tested and developed a protocol that emulates cryo-SEM using standard SEM equipment and, importantly, that results in minimal tissue damage.

Although it was developed to address specific constraints for spaceflight experiments, Paul notes that their new method is broadly applicable. "There are a lot of situations where biologists want to collect samples in extreme situations. In our case -- a space vehicle orbiting the Earth."

The new protocol maximizes the amount of data obtained from a single sample and allows for the concomitant examination of both molecular and morphological features. Using a single fixation protocol, direct comparisons between changes in morphology and altered gene expression can be made. Such an analysis not only makes full use of samples and replicates but also enables a robust analysis of the relationship between heredity and development. "Putting two tools together, it is powerful to look at the morphology in conjunction with the genes that are being expressed," says Paul.

The new protocol boasts low costs and high accessibility, and has wide application to any situation where recovery of biological resources is limited. Notably, this includes researchers collecting and preserving samples in the field, where space for materials is at a premium. "In places where sampling is limited, difficult, or expensive, the use of preservatives allows for more routes to analysis," notes Ferl.

View the original article here