The protocol begins with a basic dissection kit used to isolate the grasshopper guts. A DNA extraction is then performed on the gut components, which results in a combination of grasshopper and plant DNA. Isolating the plant DNA involves a simple polymerase chain reaction, or PCR, which is used to amplify desired regions of genetic material for further research.
A major advantage of this method is that it can be completed in less than three hours and utilizes inexpensive laboratory equipment accessible to researchers with less funding. It also includes a new technique to divide the gut into sections, enabling researchers to track the step-by-step movement of plant matter through each gut compartment.
"We can follow plant food movement during its consumption, record the sequence of food digested (what plant was chosen to consume first) or the time needed for food digestion in each compartment, and ultimately better understand the insect food digestion process," Avanesyan explains. "It opens doors to a completely different research areainsect physiology."
To demonstrate the utility of the protocol, Avanesyan successfully amplified the DNA of a noncoding region of a plant chloroplast gene and performed multiple feeding trials. Results indicated that plant tissue could be detected up to 12 hours after ingestion in nymph M. differentialis and M. bivittatus grasshoppers and adult M. femurrubrum grasshoppers. For adult M. differentialis grasshoppers, which were the largest in size, plant tissue was detected up to 22 hours post-ingestion. This information lets researchers know how to time the dissection with feeding experiments.
Findings from the gut separation technique uncovered interesting details about M. differentialis grasshoppers. They often did not switch between grasses during feeding, but instead consumed different plant species sequ
|Contact: Beth Parada|
American Journal of Botany