Real-time PCR for identification of five species of Cryptolestes based on COI barcode region

Cryptolestes: (Coleoptera: Laemophloeidae) are economically important pests, which damage stored products in granaries and mills. Because it is difficult to distinguish Cryptolestes species morphologically, we develop here a high sensitivity real-time PCR method for distinguishing five species of Cryptolestes (C. ferrugineus, C. pusillus, C. turcicus, C. pusilloides and C. capensis). The lower limit of DNA concentration required for species identification is 0.1 ng/mu l for C. ferruginues and C. pusilloides, and 0.01 ng/mu l for C. pusillus, C. turcicus, and C. capensis. This method successfully determined species assignments for previously unidentified Cryptolestes spp. Collected from the field and will facilitate Cryptolestes identification in the field of plant quarantine and stored product protection. (C) 2020 Elsevier Ltd. All rights reserved.