Jump to ContentJump to Main Navigation
Show Summary Details
More options …

DNA Barcodes

Ed. by Mitchell, Andrew

1 Issue per year

Emerging Science

Open Access
See all formats and pricing
More options …

COI gene geographic variation of Gypsy moth (Lepidoptera: Lymantriidae) and a TaqMan PCR diagnostic assay

Lu Qian / Yulin An / Junxian Song / Mei Xu / Jianlin Ye / Cuiping Wu / Bin Li / Dejun Hao
Published Online: 2014-04-24 | DOI: https://doi.org/10.2478/dna-2014-0002


Gypsy moth, an important forest/urban pest worldwide, is separated into the European and Asian subspecies, and has important quarantine significance. Diagnostic technique that can accurately and quickly distinguish subspecies is lacking. This study aimed to evaluate genetic difference between the subspecies, and subsequently to develop a reliable and high throughput molecular based diagnostic tool for distinguishing the subspecies. COI genes of 25 gypsy moth samples from China, Russia, Mongolia, Japan and the United States were sequenced. DNASTAR analysis revealed that gypsy moth COI gene was 1531bp long. The UPGM phylogenetic tree constructed based on the COI gene indicated that European subspecies (U.S. population) and Asian subspecies were distinctively divided into two branches. Japanese populations had a far distantly relationship with other Asian populations forming a separate branch. There was a single base substitution (base transformation only) at 14 consistent locations between Asian and American populations, but 13 of them coded the same amino acid. A MGB proper and TaqMan assay was designed base on the base substitution at 406th bp that coded a different amino acid. This allele typing assay took only 4 hours and could accurately distinguish gypsy moth subspecies of Europe and Asia. The study enriches the knowledge basis of genetic differentiation of gypsy moth subspecies. And more importantly the TaqMan assay is the first report of such diagnostic tool that could deliver rapid and accurate results and suitable for routine quarantine inspections to distinguish Asian and European gypsy moth subspecies. This study was supported by the Ministry of Science and Technology of the People’s Republic of China (Science and technology supporting project: 2012BAK11B03; International cooperation project: 2009DFA31950) and Jiangsu Entry and Exit Inspection and Quarantine Bureau (2014KJ45).

Keywords : Gypsy moth subspecies; COI gene; molecular marker; TaqMan assay


  • [1] Leonard D. E., Recent developments in ecology and control of the gypsy moth, Ann. Rev. Entomol. 1974, 19, 197-229.CrossrefGoogle Scholar

  • [2] Pogue M. G., Schaefer P. W., A review of selected species of Lymantria including three new species. USDA, Forest Health Technology Enterprise Team, 2007, 11-36.Google Scholar

  • [3] Bogdanowicz S.M., Wallner E.E., Bell J., O’Dell T.M., Harrision R.G. Asian gypsy moths (Lepidoptera: Lymantriidae) in North America: Evidence from molecular data, Ann. Entomol. Soc. Am. 1993, 86, 710-715.Google Scholar

  • [4] Bogdanowicz S.M., Mastro V. C., Prasher D. C, Harrison R. G., Microsatellite DNA variation among Asian and North American gypsy moths (Lepidoptera: Lymantriidae), Ann. Entomol. Soc.Am. 1997, 90, 768-775.Google Scholar

  • [5] Pfeifer T.A., Humble L.M., Ring M., Grigliatti T.A., Characterization of gypsy moth populations and related species using a nuclear DNA marker, Can. Entomol, 1995, 127, 49-58.Google Scholar

  • [6] Schreiber D.E., Garner K.J., Slavicek J.M., Identification of three randomly amplified polymorphic DNA polymerase chain reaction markers for distinguishing Asian and North American gypsy moths (Lepidoptera: Lymantriidae). Ann. Entomol. Soc.Am., 1997, 90, 667-674.Google Scholar

  • [7] Reineke A., Karlovsky P., Zebitz C. P. W., Amplified fragment length polymorphism analysis of different geographic populations of the gypsy moth, Lymantria dispar (Lepidoptera: Lymantriidae), Bull. Entomol. Res., 1999, 89, 79-88.Google Scholar

  • [8] Baranchikov Y. N., Ecological basis of the evolution of host relationships in Eurasian gypsy moth populations. In: Lymantriidae: A comparison of features of New and Old World tussock moths, W. E. Wallner, K.A. McManus (Eds.), Northeastern Forest Experiment Station, General Technical Report 1988, NE-123, 319-338.Google Scholar

  • [9] Wallner W. E.; Grinberg P. S.; Keena M. A., Female flight: evaluation of Asian gypsy moth and its hybrids, Proceedings of the USDA Interagency Gypsy Moth Research Forum 1994, Northeastern Forest Experiment Station, General Technical Report NE-188, 89.Google Scholar

  • [10] deWaard J.R., Mitchell A., Keena M. A, Gopurenko D., Boykin L. M., Armstrong K. F., et al., Towards a global barcode library for Lymantria (Lepidoptera: Lmantriinae) tussock moths of biosecurity concern, PLoS One 2010, 5, e14280, DOI:10.1371/ journal.pone.0014280. CrossrefWeb of ScienceGoogle Scholar

  • [11] Garner K.J., Slavicek J.M., Identification and characterization of a RAPD-PCR marker for distinguishing Asian and North American gypsy moths, Insect Mol. Biol., 1996, 5, 81-91.PubMedCrossrefGoogle Scholar

  • [12] Smith P.J., Mcveagh S.M., Won Y., Vrijenhoeack R. C., Genetic heterogeneity among New Zealand species of hydrothermal vent mussels (Mytilidae: Bathymodiolus), Mar. Biol., 2004, 4, 537-545.CrossrefGoogle Scholar

  • [13] Donald K.M., Kenned Y.M., Spencher H.G., The phylogeny and taxonomy of austral monodontine top shells (Mollusca: Gastropoda, Trochidae) inferred from DNA sequences, Mol.Phylogen. Evol., 2005, 37, 474- 483.CrossrefGoogle Scholar

  • [14] Hebert P. D. N., Cywińska A., Ball S.L., deWaard J.R., Biological identifications through DNA barcodes, Proc. R. Soc. Lond. B 2003, 270, 313-321.Google Scholar

  • [15] Ptaszyńska A.A., Łętowski J., Gnat S., Małek W., Application of COI sequences in studies of phylogenetic relationships among 40 Apionidae species, J. Insect Sci., 2012, 12, 16, DOI: 10.1673/031.012.1601.CrossrefGoogle Scholar

  • [16] Harrision R.C. Winterneyer S. F., Odell T.M.,Patterns of genetic variation whthin and among gypsy moth, Lymantria dispar (Lepidoptera: Lymantriidae) populations, Ann. Entomol. Soc.Am., 1983, 76, 652-656.Google Scholar

  • [17] Bogdanowicz S.M., Schaefer P. W., Harrison, R.G., Mitochondrial DNA variation among worldwide populations of gypsy moths, Lymantria dispar, Mol. Phylogen. Evol., 2010, 15, 487-495.Google Scholar

  • [18] Kavita S. L., Vidya A. A., Quantitation of hepatitis B virus DNA by real-time PCR using internal amplification control and dual TaqMan MGB probes, J Virol. Methods. 2006, 135, 83-90.Google Scholar

  • [19] Malmström S., Hannoun C., Lindh M., Mutation analysis of lamivudine resistant hepatitis B virus strains by TaqMan PCR, J.Virol. Methods. 2007, 143, 147-152.Web of ScienceGoogle Scholar

  • [20] Pholampaisathit S., Real-time Diagnosis and Monitoring of Cytomegalovirus Infections Using TaqMan-MGB Probe Technology. Int. J. Infectious Diseases, 2008, 12, Suppl. 1, 471-472. Web of ScienceGoogle Scholar

About the article

Received: 2013-11-27

Accepted: 2014-02-23

Published Online: 2014-04-24

Published in Print: 2014-01-01

Citation Information: DNA Barcodes, Volume 2, Issue 1, ISSN (Online) 2299-1077, DOI: https://doi.org/10.2478/dna-2014-0002.

Export Citation

© 2014 Lu Qian et al.. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. BY-NC-ND 3.0

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

Donald Stewart, Reza Zahiri, Abdelmadjid Djoumad, Luca Freschi, Josyanne Lamarche, Dave Holden, Sandra Cervantes, Dario I. Ojeda, Amélie Potvin, Audrey Nisole, Catherine Béliveau, Arnaud Capron, Troy Kimoto, Brittany Day, Hesther Yueh, Cameron Duff, Roger C. Levesque, Richard C. Hamelin, Michel Cusson, and Ruslan Kalendar
PLOS ONE, 2016, Volume 11, Number 8, Page e0160878
M. S. Islam, N. B. Barr, W. E. Braswell, M. Martinez, L. A. Ledezma, J. Molongoski, V. Mastro, and E. L. Schuenzel
Journal of Economic Entomology, 2015, Volume 108, Number 5, Page 2450

Comments (0)

Please log in or register to comment.
Log in