University of Oxford Oxford, England, United Kingdom
Several 1000 specimens belonging to Encarsia, Megaphragma and related genera, collected from all biogeographic regions, were extracted for genomic DNA using a technique that retains the exoskeleton complete. Although these genera include some of the smallest insects known, This protocol results in sufficient DNA to perform almost unlimited sequencing, including whole genome, while retaining a perfectly intact specimen for morphological study and permanent storage as a voucher. Both ribosomal (28S D2-D3) and mitochondrial (CO1) Sanger sequencing and analysis reveal large numbers of cryptic species that are so far indistinguishable using morphology. A further benefit of the protocol is to identify as conspecific specimens from extremely diverse localities whose conspecificity based solely on morphology would otherwise be called into question. One particularly unusual example is Megaphragma longiciliatum recorded (based on DNA) from D. R. Congo, France, Indonesia (Java), Italy, Malaysia (Borneo, Sarawak) and Oman.
