title: A Multi-Species TaqMan PCR Assay for the Identification of Asian Gypsy Moths (Lymantria spp.) and Other Invasive Lymantriines of Biosecurity Concern to North America.
abstract: Preventing the introduction and establishment of forest invasive alien species (FIAS) such as the Asian gypsy moth (AGM) is a high-priority goal for countries with extensive forest resources such as Canada. The name AGM designates a group of closely related Lymantria species (Lepidoptera: Erebidae: Lymantriinae) comprising two L. dispar subspecies (L. dispar asiatica, L. dispar japonica) and three closely related Lymantria species (L. umbrosa, L. albescens, L. postalba), all considered potential FIAS in North America. Ships entering Canadian ports are inspected for the presence of suspicious gypsy moth eggs, but those of AGM are impossible to distinguish from eggs of innocuous Lymantria species. To assist regulatory agencies in their identification of these insects, we designed a suite of TaqMan® assays that provide significant improvements over existing molecular assays targeting AGM. The assays presented here can identify all three L. dispar subspecies (including the European gypsy moth, L. dispar dispar), the three other Lymantria species comprising the AGM complex, plus five additional Lymantria species that pose a threat to forests in North America. The suite of assays is built as a "molecular key" (analogous to a taxonomic key) and involves several parallel singleplex and multiplex qPCR reactions. Each reaction uses a combination of primers and probes designed to separate taxa through discriminatory annealing. The success of these assays is based on the presence of single nucleotide polymorphisms (SNPs) in the 5' region of mitochondrial cytochrome c oxidase I (COI) or in its longer, 3' region, as well as on the presence of an indel in the "FS1" nuclear marker, generating North American and Asian alleles, used here to assess Asian introgression into L. dispar dispar. These assays have the advantage of providing rapid and accurate identification of ten Lymantria species and subspecies considered potential FIAS.
authors: Laurentian Forestry Centre, Canadian Forest Service, Natural Resources Canada, Quebec City, Quebec, Canada.
authors: Donald
authors: D
authors: Stewart
authors: Canadian Food Inspection Agency, Ottawa Plant Laboratory, Entomology Unit, Ottawa, Ontario, Canada.
authors: Reza
authors: R
authors: Zahiri
authors: Laurentian Forestry Centre, Canadian Forest Service, Natural Resources Canada, Quebec City, Quebec, Canada.
authors: Abdelmadjid
authors: A
authors: Djoumad
authors: Institut de Biologie Intégrative et des Systèmes, Université Laval, Quebec City, Quebec, Canada.
authors: Luca
authors: L
authors: Freschi
authors: Laurentian Forestry Centre, Canadian Forest Service, Natural Resources Canada, Quebec City, Quebec, Canada.
authors: Josyanne
authors: J
authors: Lamarche
authors: Canadian Food Inspection Agency, Burnaby, British Columbia, Canada.
authors: Dave
authors: D
authors: Holden
authors: Department of Forest and Conservation Sciences, Faculty of Forestry, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Sandra
authors: S
authors: Cervantes
authors: Department of Forest and Conservation Sciences, Faculty of Forestry, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Dario I
authors: DI
authors: Ojeda
authors: Laurentian Forestry Centre, Canadian Forest Service, Natural Resources Canada, Quebec City, Quebec, Canada.
authors: Amélie
authors: A
authors: Potvin
authors: Laurentian Forestry Centre, Canadian Forest Service, Natural Resources Canada, Quebec City, Quebec, Canada.
authors: Audrey
authors: A
authors: Nisole
authors: Laurentian Forestry Centre, Canadian Forest Service, Natural Resources Canada, Quebec City, Quebec, Canada.
authors: Catherine
authors: C
authors: Béliveau
authors: Department of Forest and Conservation Sciences, Faculty of Forestry, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Arnaud
authors: A
authors: Capron
authors: Canadian Food Inspection Agency, Burnaby, British Columbia, Canada.
authors: Troy
authors: T
authors: Kimoto
authors: Canadian Food Inspection Agency, National Headquarters, Ottawa, Ontario, Canada.
authors: Brittany
authors: B
authors: Day
authors: Department of Forest and Conservation Sciences, Faculty of Forestry, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Hesther
authors: H
authors: Yueh
authors: Canadian Food Inspection Agency, National Headquarters, Ottawa, Ontario, Canada.
authors: Cameron
authors: C
authors: Duff
authors: Institut de Biologie Intégrative et des Systèmes, Université Laval, Quebec City, Quebec, Canada.
authors: Roger C
authors: RC
authors: Levesque
authors: Institut de Biologie Intégrative et des Systèmes, Université Laval, Quebec City, Quebec, Canada.|Department of Forest and Conservation Sciences, Faculty of Forestry, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Richard C
authors: RC
authors: Hamelin
authors: Laurentian Forestry Centre, Canadian Forest Service, Natural Resources Canada, Quebec City, Quebec, Canada.|Institut de Biologie Intégrative et des Systèmes, Université Laval, Quebec City, Quebec, Canada.
authors: Michel
authors: M
authors: Cusson
mesh_terms: D000818:Animals; D003576:Electron Transport Complex IV / Q000737:chemistry; D005819:Genetic Markers; D058865:Introduced Species; D009036:Moths / Q000145:classification / Q000235:genetics*; D009656:North America; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D013045:Species Specificity
chemical_list: D005819:Genetic Markers; D003576:Electron Transport Complex IV