C. S.

Biology Lab Signature Assignment

Abstract: Visualization of Extracellular DNA Released During Boarder Cell Separation

from the Root Cap

Premise of the study:

This experiment was performed to help provide a visualization of what happens to the

extracellular DNA (border cells) in root caps when it touches water. Border cells are

cells that actively release DNA proteins that separate and function a defense system for

plants. The specific DNA they were studying was very small, amounts greater than 100

proteins would be secreted, and they generally were secreted during the release from

the root tip. These border cells help combat intruders from gaining access to the cell,

just like a mammal’s white blood cell would trap infection. They do this by telling the cell

to trap the foreign object. The researches wanted to identify the role of DNA in the

structural integrity of these extracellular proteins that are released as the border cells

dispersed from the root tip and encountered water.

Methods:

In order to measure and observe the particulates in the experiment the researchers had

to use stains to coat the different parts that they predicted to see. The stains they used

comprised of; crystal violet, toluidine blue, Hoechst 33342, DAPl, and SYTOX green.

Each stain was dropped onto the root tip to highlight the extracellular viscous solution

secreted as it absorbed water and began to dissipate. To evaluate the changes in the
structure researchers used a product called DNase I. This material allowed for the easy

viewing of the extracellular DNA after it started to breakdown.

Results:

Researchers concluded that complex masses of border cells were instantly present

when they came into contact with the stains. Even stains that only coat DNA were

visible. The samples that had DNA were then treated with DNase I. The treatment

greatly transformed the appearance of the structure, so much so, that the association

with the border cells had been damaged. To confirm the findings researchers ran the

same stain tests on border cells that had been rendered useless due to freezing and

being run through a centrifuge. When the stains were added in these instances, the

DNA was not observable. The results from these experiments helped prove the

hypothesis that DNA is only secreted by living cells.

Conclusion:

Results drawn from this experiment ensure that DNA is an important part of the border

cell extracellular traps. Most plant cells have these cells that are coded to release from

the root tip when it is growing in soil. The protein delivery was confirmed to be an active

process by the comparison to the cells already killed by freezing and centrifuge. An

added discovery during the experiment came when Histone secretions lead Zychlinsky

Laboratory in 2004 to report its findings of neutrophils to export histone-linked

extracellular DNA as part of the structures that immobilize pathogens and stop infection.

This statement means that there is a parallel to animal systems. When the DNase I was

added to the traps in the animal cells the traps opened, and the infection was released.
Subsequently, when the DNase I was added to the root tip cells they lost the ability to

fight infection at all.

Reflection:

Although the article was a bit advanced for my level of understanding in the field of

study, it gave me a new appreciation for the hard work that the members of the botany

community are putting in. Their work provides us a chance to learn about the survival of

not only the different plants that we depend on, but our mammal cells as well. The

ability to have a defense system is relied upon immensely for living organisms. This

study helped offer a bridge that links the way our cells trap infection to the way that

plant cells root caps to do the same. It is exciting to learn that there is a correlation

between our cell structures and plant cell structures for the specific purpose of

protection.

Bibliography:

Date accessed March 2018:

Visualization of extracellular DNA released during border cell separation from the

root cap