Molecular Diagnostics of Enaphalodes rufulus (Coleoptera: Cerambycidae)
Abstract
Oak-hickory forests in northwestern Arkansas, eastern Oklahoma and southern Missouri have recently experienced an oak decline event with widespread oak mortality. The oak mortality is associated with an outbreak of a native wood-boring cerambycid, Enaphalodes rufulus (Haldeman), the red oak borer. Taxonomic identification, below the family level, of larval Cerambycidae through traditional morphological methods is not usually possible. We employed molecular diagnostics, with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), to distinguish E. rufulus from other closely related species of cerambycids. A portion of the mitochondrial DNA 16S rRNA gene, isolated from legs or thoraxes of adult museum specimens, was amplified and digested with Alu I and Hind III restriction enzymes. Both restriction enzymes independently produced fragments for E. rufulus that were significantly different from any other cerambycid tested. Alu I had one restriction site for E. rufulus and two restriction sites for all other cerambycids tested, while Hind III did not cut for E. rufulus but did cut at one restriction site for all other cerambycids. Eggs, larvae, and pupae of E. rufulus along with an unknown cerambycid larva and pupa were successfully amplified and digested by this method to verify validity of this technique for multiple life stages.View this article in BioOne
Downloads
Published
Issue
Section
License
Copyright for any article published in Florida Entomologist is held by the author(s) of the article. Florida Entomologist is an open access journal. Florida Entomologist follows terms of the Creative Commons, Attribution Non-Commercial License (cc by-nc). By submitting and publishing articles in Florida Entomologist, authors grant the FOJ and Florida Entomologist's host institutions permission to make the article available through Internet posting and electronic dissemination, and to otherwise archive the information contained both electronically and in a hard printed version. When used, information and images obtained from articles must be referenced and cited appropriately. Articles may be reproduced for personal, educational, or archival purposes, or any non-commercial use. Permission should be sought from the author(s) for multiple, non-commercial reproduction. Written permission from the author(s) is required for any commercial reproduction.