Coupling scanning electron microscopy with DNA bar coding: a novel approach for thrips identification

The small size and cryptic nature of thrips pests often make their monitoring and the identification process difficult. With the aim to obtain accurate identification of various pest species in the order Thysanoptera, a pilot study was conducted on an invasive thrips species Scirtothrips dorsalis Hood in Florida. The specific objective of this pilot study was to assess if the thrips specimens processed to be observed under scanning electron microscopy (SEM) can be further utilized for DNA based assays. Larvae and adults of S. dorsalis were subjected to traditional morphological identification using high resolution SEM prior to their DNA extraction. Sequence results of both mtCO1 and ITS rDNA of individual larva and adult S. dorsalis were found to be in agreement with the taxonomic identification conducted using SEM and each result confirmed the other technique. Our results suggest that steps involved during specimen preparation and observation under SEM does not impact DNA analysis of the sample. The two techniques together could be used for the correct identification of various thrips species using the same specimen. The proof of concept was further tested on three other important thrips species Frankliniella occidentalis (Pergande), F. schultzei (Trybom) and Thrips palmi Karny, which confirmed the robustness of the technique. The technique has broader significance for different areas under entomological sciences especially medical, forensic and taxonomic studies where the samples put under a high beam of ions for investigation are often required for DNA analysis.