从蚱蜢肠道中提取植物 DNA：从组织准备到获得植物 DNA 序列的分步方案。
Plant DNA detection from grasshopper guts: A step-by-step protocol, from tissue preparation to obtaining plant DNA sequences.
机构信息
Department of Biological Sciences, University of Cincinnati, 614 Rieveschl Hall, Cincinnati, Ohio 45221-0006 USA.
出版信息
Appl Plant Sci. 2014 Feb 1;2(2). doi: 10.3732/apps.1300082. eCollection 2014 Feb.
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.
研究前提
开发了一种基于 PCR 的方法，用于鉴定食用黑麦草蝽肠道内容物中的植物 DNA。尽管之前的研究集中在各种昆虫上，但目前还没有针对农业害虫——黑麦草蝽的植物 DNA 检测协议，而这些害虫会显著影响植物群落组成。
方法和结果
开发的协议成功使用了叶绿体 trnL（UAA）基因的非编码区，并在多个喂养实验中进行了测试。从整个肠道和肠道的不同部分在摄入后七个时间点获得了植物 DNA，在若虫中可检测到摄入后 12 小时，在成虫中可检测到摄入后 22 小时。
结论
所提出的协议是一种从黑麦草蝽肠道及其不同部位检测植物 DNA 的有效、相对快速且低成本的方法。这具有重要的应用价值，从探索植物在进食过程中的“移动”，到检测植物-昆虫相互作用。
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