TY - JOUR

T1 - A TaqMan Assay for the Detection and Monitoring of Potentially Invasive Lasiocampids, With Particular Attention to the Siberian Silk Moth, Dendrolimus sibiricus (Lepidoptera: Lasiocampidae)

AU - Stewart, Don

AU - Djoumad, Abdelmadjid

AU - Holden, Dave

AU - Kimoto, Troy

AU - Capron, Arnaud

AU - Dubatolov, Vladimir V.

AU - Akhanaev, Yuriy B.

AU - Yakimova, Maria E.

AU - Martemyanov, Vyacheslav V.

AU - Cusson, Michel

N1 - We are grateful to A. Naaum (NatureMetrics, Guelph, Canada) for insightful suggestions regarding the development of the on-site version of our assay. We thank colleagues who provided insect specimens for this study; their names are listed in ]. This work was supported by a grant from the Genomics Research and Development Initiative (Government of Canada) to MC and by grants from the Russian Science Foundation to V. V. M. and V. V. D. The work reported here was completed in 2021.

PY - 2023/1/1

Y1 - 2023/1/1

N2 - The Siberian silk moth, Dendrolimus sibiricus Tschetverikov, is a very serious pest of conifers in Russia and is an emerging threat in North America where an accidental introduction could have devastating impacts on native forest resources. Other Dendrolimus Germar species and related Eurasian lasiocampids in the genus Malacosoma (Hubner) could also present a risk to North America's forests. Foreign vessels entering Canadian and U.S. ports are regularly inspected for Lymantria dispar (Linnaeus) and for the presence of other potentially invasive insects, including suspicious lasiocampid eggs. However, eggs are difficult to identify based on morphological features alone. Here, we report on the development of two TaqMan (Roche Molecular Systems, Inc., Rotkreuz, Switzerland) assays designed to assist regulatory agencies in their identification of these insects. Developed using the barcode region of the cytochrome c oxidase I (COI) gene and run in triplex format, the first assay can detect Dendrolimus and Malacosoma DNA, and can distinguish North American from Eurasian Malacosoma species. The second assay is based on markers identified within the internal transcribed spacer 2 (ITS2) region and was designed to specifically identify D. sibiricus, while discriminating closely related Dendrolimus taxa. In addition to providing direct species identification in the context of its use in North America, the D. sibiricus assay should prove useful for monitoring the spread of this pest in Eurasia, where its range overlaps with those of the morphologically identical D. superans (Butler) and similar D. pini (Linnaeus). The assays described here can be performed either in the lab on a benchtop instrument, or on-site using a portable machine.

AB - The Siberian silk moth, Dendrolimus sibiricus Tschetverikov, is a very serious pest of conifers in Russia and is an emerging threat in North America where an accidental introduction could have devastating impacts on native forest resources. Other Dendrolimus Germar species and related Eurasian lasiocampids in the genus Malacosoma (Hubner) could also present a risk to North America's forests. Foreign vessels entering Canadian and U.S. ports are regularly inspected for Lymantria dispar (Linnaeus) and for the presence of other potentially invasive insects, including suspicious lasiocampid eggs. However, eggs are difficult to identify based on morphological features alone. Here, we report on the development of two TaqMan (Roche Molecular Systems, Inc., Rotkreuz, Switzerland) assays designed to assist regulatory agencies in their identification of these insects. Developed using the barcode region of the cytochrome c oxidase I (COI) gene and run in triplex format, the first assay can detect Dendrolimus and Malacosoma DNA, and can distinguish North American from Eurasian Malacosoma species. The second assay is based on markers identified within the internal transcribed spacer 2 (ITS2) region and was designed to specifically identify D. sibiricus, while discriminating closely related Dendrolimus taxa. In addition to providing direct species identification in the context of its use in North America, the D. sibiricus assay should prove useful for monitoring the spread of this pest in Eurasia, where its range overlaps with those of the morphologically identical D. superans (Butler) and similar D. pini (Linnaeus). The assays described here can be performed either in the lab on a benchtop instrument, or on-site using a portable machine.

KW - Dendrolimus

KW - Lasiocampidae

KW - Malacosoma

KW - TaqMan

KW - molecular identification

KW - Canada

KW - Animals

KW - Insecta

KW - Moths/genetics

KW - Ovum

KW - Manduca

KW - Bombyx

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85147318047&origin=inward&txGid=cf1512d94717988595c0800e29f2d383

UR - https://www.mendeley.com/catalogue/f573ca67-f0d7-35f4-adf9-6c7381b571d5/

U2 - 10.1093/jisesa/ieac062

DO - 10.1093/jisesa/ieac062

M3 - Article

C2 - 36723233

VL - 23

JO - Journal of insect science

JF - Journal of insect science

SN - 1536-2442

IS - 1

M1 - 5

ER -