title: A molecular tool for detection and tracking of a potential indigenous Beauveria bassiana strain for managing emerald ash borer populations in Canada.
abstract: Emerald ash borer is an invasive species from Asia. Beauveria bassiana strain L49-1AA is being tested for the control of emerald ash borer in Canada, using an autocontamination trapping system. We have developed a simplified allele discrimination polymerase chain reaction (PCR) assay to screen B. bassiana strain, L49-1AA from other Beauveria species by targeting the inter-strain genetic differences in 5' end of EF1-α gene of the genus Beauveria. A single nucleotide polymorphism (SNP) site, T→C was identified only in L49-1AA and was used to develop a simplified allele discrimination polymerase chain reaction (PCR) assay based on a modified allelic inhibition of displacement activity (AIDA) approach for distinguishing B. bassiana L49-1AA from all background Beauveria isolates. The SNP site was employed to design inner primers but with a deliberate mismatch introduced at the 3' antepenultimate from the mutation site in order to maximize specificity and detection efficiency. Amplification was specific to L49-1AA without cross-reaction with DNA from other Beauveria strains. In addition, the designed primers were also tested against environmental samples in L49-1AA released plots and observed to be highly efficient in detecting and discriminating the target strain, L49-1AA from both pure and crude DNA samples. This new method can potentially allow for more discriminatory tracking and monitoring of released L49-1AA in our autocontamination and dissemination projects for managing EAB populations. Additionally, the modified-AIDA format has potential as a tool for simultaneously identifying and differentiating closely related Beauveria species, strains/isolates as well as general classification of other pathogens or organisms.
authors.affiliation: Canadian Forestry Service, Great Lakes Forestry Centre, Natural Resources Canada, 1219 Queen Street East, Sault Ste. Marie, Ontario P6A 2E5, Canada.
authors.forename: Shajahan
authors.initials: S
authors.lastname: Johny
authors.affiliation: Canadian Forestry Service, Great Lakes Forestry Centre, Natural Resources Canada, 1219 Queen Street East, Sault Ste. Marie, Ontario P6A 2E5, Canada. Electronic address: gkyeipok@NRCan.gc.ca.
authors.forename: George
authors.initials: G
authors.lastname: Kyei-Poku
mesh_terms: D000818:Animals; D052982:Beauveria / Q000235:genetics / Q000302:isolation & purification*; D002170:Canada; D001517:Coleoptera / Q000382:microbiology*; D031661:Fraxinus / Q000382:microbiology; D010572:Pest Control, Biological / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
keywords: Beauveria bassiana; Biological control; Emerald ash borer; Molecular marker