Identification of leaf-mining insects via DNA recovered from empty mines

Abstract

We report the development of an improved method for the extraction and amplification of leaf miner DNA recovered from empty mines. Our method is simple, easy to use, and foregoes the time-consuming task of scraping out mines required by previous methods. We collected leaves with 1- and 2-day-old vacated mines, cut out and then ground the mined portions, and amplified the mtDNA COI barcode sequence using universal insect primers. We obtained high-quality sequences for 31% of our empty mines: 20% yielded sequences associated with a leaf miner species; and an additional 11% yielded sequences associated with whiteflies, mites, or fungi. Our improved method will facilitate ecological studies determining herbivore community dynamics and agricultural studies for pest monitoring and identification.