Identification of morphologically similar canegrubs (Coleoptera : Scarabaeidae : Melolonthini) using a molecular diagnostic technique

Larvae of 19 species of Melolonthini (Coleoptera: Scarabaeidae) are importa nt sugarcane pests in eastern Australia. Larvae of five of these pest speci es cannot be identified confidently using morphological characters: Lepidio ta frenchi Blackburn, Lepidiota negatoria Blackburn and Lepidiota noxia Bri tton; and Antitrogus consanguineus (Blackburn) and Antitrogus rugulosus (Bl ackburn). To develop a DNA-diagnostic tool, we assessed mtDNA diversity wit hin each species and confirmed their specific status. Species-level base pa ir differences in the cytochrome oxidase II gene (COII) were used to develo p a polymerase chain reaction-restriction fragment length polymorphism (PCR -RFLP) protocol for species identification. PCR-RFLP is a useful tool for i dentifying morphologically similar species of canegrubs in large-scale pest identifications.