Molecular identification of elm bark beetles through DNA barcoding

Document Type : Research Paper

Authors

1 Postdoctoral Research Department of Plant Protection, College of Agriculture and Natural Resources,

2 Associate Professor Department of Plant Protection Faculty of Agronomy Sciences College of Agriculture & Natural Resources, University of Tehran

3 Department of plant protection, Faculty of Agricultural sciences, University of Guilan.

Abstract

Bark beetles as one of the most important pest group attacked to weak trees in forests and urban trees. Ulmus minor considers as an important host plant for bark beetles. This group of beetles feed on Phloem and transmit the pathogen of Dutch elm disease that cause high damage in Ulmus trees. The first and the most important step in preventing damage and bark beetles management is accurate identification of bark beetles. Because of small size and similarity in morphological characters of bark beetles, identification of this group is difficult and time consuming. In this study a DNA-based method barcoding was used to identify six bark beetles species that collected on Ulmus minor through sequencing 580bp of cytochrome oxidase one genome by using A2411 and C-N-J1718 universal primers. Obtained sequences were analyzed by MEGA ver.0 software. The results showed that the mean of interspecies nucleotide variation is 1.9% more than intraspecific variation 0.03% which confirmed DNA barcoding rule. The result showed that specimen identified in species level which proved DNA barcoding as a reliable tool to identify Ulmus bark beetles species.

Keywords


  1. Amini, S. & Hosseini, R. (2012). Introduction and morphological identification key for three elm bark beetle species in Guilan province. Plant Pests Research. 3, 13-20.(In farsi)
  2. Amini, S. & Hosseini, R. (2015). A multiplex polymerase chain reaction based method for rapid identification of two species of the genus Scolytus Geoffroy (Col: Curculionidae: Scolytinae) in Iran. Journal of Entomological and Acarological research, 48, 51-81.
  3. Caterino, M. S., S. Cho & Sperling, F. A. H (2000). The current state of insect molecular systematics: a thriving tower of babel. Annual Review of Entomology. 45: 1-54.
  4. Cognato, A.I. & Sun. J.H. (2007). DNA based cladograms augment the discovery of a new Ips species from China (Coleoptera: Curculionidae: Scolytinae). Cladistics. 23: 539-551.
  5. Cognato, A. I. & Sperling, F. A. H. (2000). Phylogeny of Ips DeGeer species (Coleoptera: Scolytidae) inferred from mitochondrial cytochromeoxidase I sequence. Molecular Phylogenetics and Evolution, 14, 445–460.
  6. Gregoire, J.C. & Evans, H.F. (2004) Damage and control of BAWBILT organisms, an overview. Bark and Wood Boring Insects in Living Trees in Europe, a Synthesis, 24:19-37.
  7. Hajibabaei, M., Janzen, J. M., Burns, D. H., Hallwachs, W. & Hebert, P. D. N. (2006). DNA barcodes distinguish species of tropical Lepidoptera.In Proceeding of the National Academy of Science, 103, 968-971.
  8. Hebert, P.D.N., A. Cywinska, S.L. Ball & J.R. de Waard. (2003). Biological identification through DNA barcodes. In: Proceedings of the Royal Society B-Biological Sciences. 270: 313-321.
  9. Hosseini, R. (2010) An introduction to the principle of molecular biology techniques (With emphasizing on the study of insects). University of Guilan Press. 232 pp (In Farsi).
  10. Jackson, P. L., Straussfogel, D., Lindgren, B. S., Mitchell, S. & Murphy, B. D. (2008). Radar observation and aerial capture of mountain pine beetle, Dendroctonus ponderosae (Coleoptera: Scolytidae) in flight above the forest canopy. Canadian Journal of Forest Research, 38, 2313–2327.
  11. Jordal, B.H., B.B. Normark, & Farrell, B.D. (2000). Evolutionary radiation of an inbreeding haplodiploid beetle lineage, (Curculionidae, Scolytinae). Biological Journal of the Linnean Society, 71: 499-483.
  12. Jordal, B. H. & Kambestad, M. (2014). DNA barcoding of bark and ambrosia beetles reveals excessive NUMTs and consistent east-west divergence across Palearctic forests. Molecular Ecology Resources, 14: 7–17.
  13. Kelley, S. T. & Farrell, B. D. (1998). Is specialization a dead end? The phylogeny of host use in Dendroctonus bark beetles (Scolytidae). Evolution, 52, 1731–1743.
  14. Lawrence, J. F. & Newton, A. F. (1995). Families and subfamilies of Coleoptera (with selected genera, note, references and data on family grouped names). In: J., Pakaluk, S. A., Ślipiński, R. A. Crowson. Biology, Phylogeny and classification of Coleoptera. Paper celebrating the 80th birthday of Roy A. Crowson, Volume 1, Muzeum i Instytut Zoologii PAN, 2, 778-1006.
  15. Moppe, S. (1996). Adaptive genetic structure in phytophagous insect populations. Trends Ecological Evolution., 11: 235–238.
  16. Pfeffer A. & Knizek, M. (1995). Zentral und westpaläarktische Borken und Kernkäfer (Coleoptera: Scolytidae: Platypodidae). Pro Entomologia: 310 pp
  17. Roeper R.A. L.M. Treeful. K.M. O’Brien. R.A. and Bunce, M.A.(1980). Life history of the ambrosia beetle Xyleborus (Coleoptera: Scolytidae) from in vitro culture. Great Lakes Entomologist 13: 141–144.
  18. Saccaggi, D. L., Kruger, K. & Pietersen, G. (2008). A multiplex PCR assay for the simultaneous identification of three mealybug species (Hemiptera: Pseudococcidae). Bulletin of Entomological Research, 98, 27– 33.
  19. Stauffer, C, Lakatos, F, & Hewitt, G.M. (1999). Phylogeography and postglacial colonization routes of Ips typographus (Coleoptera, Scolytidae). Molecular Ecology. 8: 763–773.
  20. Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011). MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28, 2731-2739.