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Applied Entomology and Zoology

, Volume 54, Issue 1, pp 141–145 | Cite as

Rapid development and characterization of EST-SSR markers for the honey locust seed beetle, Megabruchidius dorsalis (Coleoptera: Bruchidae), using de novo transcriptome analysis based on next-generation sequencing

  • Kako OhbayashiEmail author
  • Naoko Ishikawa
  • Yoshikuni Hodoki
  • Yasukazu Okada
  • Shin-ichi Nakano
  • Motomi Ito
  • Masakazu Shimada
Technical Note
  • 46 Downloads

Abstract

We developed 10 novel simple sequence repeat markers from expressed sequence tags for the honey locust seed beetle, Megabruchidius dorsalis Fåhraeus 1839 (Coleoptera: Bruchidae, using de novo transcriptome analysis based on next-generation sequencing. In a M. dorsalis Harataima (Kanagawa-pref.) population, the number of alleles per locus ranged from 2 to 6, with an average of 4.0. Observed and expected heterozygosities ranged from 0.13 to 0.72 and 0.17 to 0.72, respectively. We initially developed 11 novel markers, but one was eliminated because it showed significant linkage disequilibrium with another locus after Bonferroni correction. To check the applicability of the remaining 10 markers, we used them to analyze two additional geographic populations (Yashima, Akita-pref., and Kameoka, Kyoto-pref.). Mean numbers of alleles per locus in the Yashima and Kameoka populations were 2.6 and 2.9, respectively, with corresponding mean observed heterozygosities of 0.36 and 0.52. These results based on the two additional populations confirm that our developed markers worked efficiently. The simple sequence repeat markers developed from expressed sequence tags in the present study should, therefore, be useful for explicating the population genetic structure of M. dorsalis and for future paternal analyses.

Keywords

EST-SSR marker A honey locust seed beetle Genetic structure Paternal analysis 

Notes

Acknowledgements

We acknowledge Dr N Nakahama (The University of Tokyo) for valuable comments and Drs H Kurota and K Takakura for sampling points. This research is partly supported by JSPS KAKENHI Grant Numbers 17H04612 to MS and 18H04815, 17H05938, 17K19381 to YO and by Joint Usage/Research Grant of Center for Ecological Research (2017jurc-cer04, 2018jurc-cer04), Kyoto University to MS.

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Copyright information

© The Japanese Society of Applied Entomology and Zoology 2019

Authors and Affiliations

  1. 1.Center for Ecological ResearchKyoto UniversityOtsuJapan
  2. 2.Department of General System Studies, Graduate School of Arts and ScienceThe University of TokyoTokyoJapan
  3. 3.Department of Biological SciencesTokyo Metropolitan UniversityTokyoJapan

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