title: Plant DNA detection from grasshopper guts: A step-by-step protocol, from tissue preparation to obtaining plant DNA sequences.
abstract: OBJECTIVE
  A PCR-based method of identifying ingested plant DNA in gut contents of Melanoplus grasshoppers was developed. Although previous investigations have focused on a variety of insects, there are no protocols available for plant DNA detection developed for grasshoppers, agricultural pests that significantly influence plant community composition. •
  
  
  RESULTS
  The developed protocol successfully used the noncoding region of the chloroplast trnL (UAA) gene and was tested in several feeding experiments. Plant DNA was obtained at seven time points post-ingestion from whole guts and separate gut sections, and was detectable up to 12 h post-ingestion in nymphs and 22 h post-ingestion in adult grasshoppers. •
  
  
  CONCLUSIONS
  The proposed protocol is an effective, relatively quick, and low-cost method of detecting plant DNA from the grasshopper gut and its different sections. This has important applications, from exploring plant "movement" during food consumption, to detecting plant-insect interactions.
authors.affiliation: Department of Biological Sciences, University of Cincinnati, 614 Rieveschl Hall, Cincinnati, Ohio 45221-0006 USA.
authors.forename: Alina
authors.initials: A
authors.lastname: Avanesyan
keywords: grasshoppers; insect gut content; plant DNA barcoding; trophic interactions