Open AccessA Multi-Species TaqMan PCR Assay for the Identification of Asian Gypsy Moths (Lymantria spp.) and Other Invasive Lymantriines of Biosecurity Concern to North America
Author(s) -
Donald J. Stewart,
Reza Zahiri,
Abdelmadjid Djoumad,
Luca Freschi,
Josyanne Lamarche,
Dave Holden,
Sandra Cervantes,
Darío I. Ojeda,
Amélie Potvin,
Audrey Nisole,
Catherine Béliveau,
Arnaud Capron,
Troy Kimoto,
Brittany Day,
Hesther Yueh,
Cameron Duff,
Roger C. Lévesque,
Richard Hamelin,
Michel Cusson
Publication year - 2016
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0160878
Subject(s) - lymantria dispar , biology , subspecies , dispar , erebidae , gypsy moth , zoology , taqman , introduced species , lepidoptera genitalia , ecology , polymerase chain reaction , genetics , gene , entamoeba histolytica
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 s pecies and subspecies considered potential FIAS.