PCR–RFLP of the mitochondrial cytochrome oxidase (subunit I) gene provides diagnostic markers for selected Diabrotica species (Coleoptera: Chrysomelidae)

Authors

Thomas L. Clark, University of Nebraska-LincolnFollow
Lance J. Meinke, University of Nebraska-LincolnFollow
John E. Foster, University of Nebraska-LincolnFollow

Date of this Version

2001

Citation

Bulletin of Entomological Research (2001) 91, pp. 419–427, DOI: 10.1079/BER2001130.

Comments

Copyright 2001 CAB International. Used by permission.

Abstract

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.