PLANT DNA DETECTION FROM GRASSHOPPER GUTS: A STEP-BY-STEP PROTOCOL, FROM TISSUE PREPARATION TO OBTAINING PLANT DNA SEQUENCES

被引:20
作者
Avanesyan, Alina [1 ]
机构
[1] Univ Cincinnati, Dept Biol Sci, Cincinnati, OH 45221 USA
来源
APPLICATIONS IN PLANT SCIENCES | 2014年 / 2卷 / 02期
关键词
grasshoppers; insect gut content; plant DNA barcoding; trophic interactions;
D O I
10.3732/apps.1300082
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Premise of the study: 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. Methods and 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.
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页数:9
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