Novel molecular identification methods for the larger black flour beetle, Cynaeus angustus (Coleoptera: Tenebrionidae)

  • Satoshi FuruiEmail author
  • Akihiro Miyanoshita
  • Taro Imamura
  • Yukio Magariyama
Technical Note


Two new methods were developed for identifying Cynaeus angustus (LeConte) (Coleoptera: Tenebrionidae) by DNA amplification using simplex and real-time PCR targeting the cytochrome c oxidase subunit I (COI) sequence reported previously. The specificities of the PCR primers and probe were also confirmed by the two PCR methods using the 22 main stored-product insect species, including DNA samples from nine tenebrionid beetle species. The results showed that the newly developed simplex and real-time PCR-based methods have sufficient specificity for analysis. The limits of detection for C. angustus total DNA by the simplex and multiplex PCR were 320 fg and 20 pg, respectively.


Real-time PCR Qualitative detection Cynaeus angustus (LeConte) Invasive insects 



We thank Dr. Kiyoshi Ando for the morphological identification of C. angustus and Ms. Miyuki Tsuru and Ms. Tomomi Endo for their diligent support to this study.


  1. Akita K, Yamaji H (2014) Cynaeus angustus found in forest field. SAYABANE N. S. 16:32 (author translated, in Japanese)Google Scholar
  2. Ando K, Ebina T (2012) Asian new record of grain-storage tenebrionid beetle, Cynaeus angustus (LeConte), intercepted at the Japanese imported plant quarantine from imported soybean waste from China. Sayabane N. S. 5:33–34 (in Japanese) Google Scholar
  3. Barak AV, Dunkel FV, Harein PK (1981) Emergence of the larger black flour beetle as a major pest of farm-stored grain in Minnesota. J Econ Entomol 74:726–729CrossRefGoogle Scholar
  4. CAC/GL 74-2010 (2010) Guidelines on performance criteria and validation of methods for detection, identification and quantification of specific DNA sequences and specific proteins in foodsGoogle Scholar
  5. Clarke LJ, Soubrier J, Weyrich LS, Cooper A (2014) Environmental metabarcodes for insects: in silico PCR reveals potential for taxonomic bias. Mol Ecol Res 14:1160–1170CrossRefGoogle Scholar
  6. Costion C, Ford A, Cross H, Crayn D, Harrington M, Lowe A (2011) Plant DNA barcodes can accurately estimate species richness in poorly known floras. PLoS One 6:e26841CrossRefGoogle Scholar
  7. Delobel A, Tran M (1993) Coleoptera of stored foodstuffs in hot regions. Faune tropicale XXXII, Orstom, Paris (in French)Google Scholar
  8. Dunkel FV, Barak AV, Harein PK (1982) Geographical distribution of Cynaeus angustus (LeConte) (Coleoptera: Tenebrionidae) and its association with stored products. J Biogeogr 9:345–352CrossRefGoogle Scholar
  9. Ferrer J (1995) A key to the flour beetles of the genus Tribolium MacLeay in Sweden (Coleoptera, Tenebrionidae), with distributional notes. Ent Tidskr 116:123–126Google Scholar
  10. Ferrer J, Andersson B (2002) Mix-up of the species of the genus Cynaeus Leconte, 1866, in Sweden and Finland. Ent Tidskr 123:219–221 (in Swedish with English abstract) Google Scholar
  11. Hatch MH (1940) Stored-grain beetles in Western Washington with special reference to the tenebrionid, Cyneus angustus LeC. The Pan-Pac Entomol 16:34–35Google Scholar
  12. Hebert PDN, Cywinska A, Ball SL, deWaard JR (2003) Biological identifications through DNA barcodes. Proc R Soc Lond B 270:313–321CrossRefGoogle Scholar
  13. Hong K, Yun T (2017) First report in Korea of the stored grain pest Cynaeus angustus (LeConte) and a pictorial identification key for tenebrionid pests in stored products. Korean J Appl Entomol 56:223–227CrossRefGoogle Scholar
  14. Ikin B, Roach A, Rees D, Banks J (1999) Pest risk analysis of a proposal for the importation of feed grain maize (Zea mays) from the USA. Arthropod pest risk analysis, CanberraGoogle Scholar
  15. Kovalenko Y, Drogvalenko AN, Khryapin RA (2016) First record of the larger black flour beetle (Cynaeus angustus) (Coleoptera: Tenebrionidae) from Russia, with a review of its North American and trans-continental expansion. Zoosyst Rossica 25:291–294CrossRefGoogle Scholar
  16. Krall JL, Decker GC (1944) The biology of Cynaeus angustus LeC., a new stored grain pest. Iowa State College J Sci 20:385–402Google Scholar
  17. LeConte JL (1852) Descriptions of new species of Coleoptera, from California. Ann N Y Acad Sci 5:125–216Google Scholar
  18. MAF Biosecurity New Zealand Standard 155.02.05 (2010) Importation of seed for sowing. Ministry of agriculture and forestry, New ZealandGoogle Scholar
  19. Nansen C, Stokes B, James J, Porter P, Shields EJ, Wheeler T, Meikle WG (2013) Biological control agent of larger black flour beetles (Coleoptera: Tenebrionidae): a nuisance pest developing in cotton gin trash piles. J Econ Entom 106:648–652CrossRefGoogle Scholar
  20. Oppert B, Walters P, Zuercher M (2006) Digestive proteinases of the larger black flour beetle, Cynaeus angustus (Coleoptera: Tenebrionidae). Bull Entomol Res 96:167–172CrossRefGoogle Scholar
  21. Ruta R, Miłkowski M, Żuk K (2017) Dacne picta Crotch, 1873 and Cynaeus angustus (LeConte, 1851)—two species new to the fauna of Poland (Coleoptera: Erotylidae, Tenebrionidae). Wiad Entomol 36:102–107Google Scholar
  22. Sambrook JF, Russell DW (2001) Molecular cloning third edition, Cold spring 349 harbor laboratory press, Cold spring harborGoogle Scholar
  23. Soldati F, Godinat G (2013) ) Cynaeus angustus (J.L. LeConte, 1851) (Coleoptera: Tenebrionidae) has been recorded in Haut-Rhin, though this species already spread through Europe. L’Entomol tome. L’Entomol tome 69:53–55Google Scholar
  24. Weston PA, Barney RJ (1998) Comparison of three trap types for monitoring insect populations in stored grains. J Econ Entomol 91:1449–1457CrossRefGoogle Scholar
  25. White NDG, Sinha RN (1987) Bioenergetics of Cynaeus angustus (Coleoptera: Tenebrionidae) feeding on stored corn. Ann Entomol Soc Am 80:184–190CrossRefGoogle Scholar
  26. Zhang T, Wang Y, Guo W, Luo D, Wu Y, Kučerová Z, Stejskal V, Opit G, Cao Y, Li F, Li Z (2016) DNA barcoding, species-specific PCR and real-time PCR techniques for the identification of six Tribolium pests of stored products. Sci Rep 6:28494CrossRefGoogle Scholar

Copyright information

© The Japanese Society of Applied Entomology and Zoology 2019

Authors and Affiliations

  • Satoshi Furui
    • 1
    Email author
  • Akihiro Miyanoshita
    • 1
  • Taro Imamura
    • 1
  • Yukio Magariyama
    • 1
  1. 1.Food Research Institute, NAROTsukubaJapan

Personalised recommendations