PCR–RFLP of the mitochondrial cytochrome oxidase (subunit I) gene provides diagnostic markers for selected \u3ci\u3eDiabrotica\u3c/i\u3e species (Coleoptera: Chrysomelidae)

Adult and larval identification of Diabrotica can be difficult. Some adult identifications require considerable taxonomic experience while larvae of many Diabrotica species are morphologically indistinguishable. This study was conducted to determine whether 12 pest and non-pest Diabrotica species could be separated using polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP). A 1308 bp portion of the mitochondrial cytochrome oxidase subunit I gene (COI) was amplified using PCR and digested using several restriction endonucleases. Double digests of COI amplicons with AluI and MspI resolved on polyacrylamide gels revealed several diagnostic inter- and intraspecific polymorphisms. A key to the 12 species was constructed using the PCR–RFLP patterns