Wolbachia sequence typing in butterflies using pyrosequencing

Sungmi Choi, Su Kyoung Shin, Gilsang Jeong, Hana Yi

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

Wolbachia is an obligate symbiotic bacteria that is ubiquitous in arthropods, with 25–70% of insect species estimated to be infected. Wolbachia species can interact with their insect hosts in a mutualistic or parasitic manner. Sequence types (ST) of Wolbachia are determined by multilocus sequence typing (MLST) of housekeeping genes. However, there are some limitations to MLST with respect to the generation of clone libraries and the Sanger sequencing method when a host is infected with multiple STs of Wolbachia. To assess the feasibility of massive parallel sequencing, also known as next-generation sequencing, we used pyrosequencing for sequence typing of Wolbachia in butterflies. We collected three species of butterflies (Eurema hecabe, Eurema laeta, and Tongeia fischeri) common to Korea and screened them for Wolbachia STs. We found that T. fischeri was infected with a single ST of Wolbachia, ST41. In contrast, E. hecabe and E. laeta were each infected with two STs of Wolbachia, ST41 and ST40. Our results clearly demonstrate that pyrosequencing-based MLST has a higher sensitivity than cloning and Sanger sequencing methods for the detection of minor alleles. Considering the high prevalence of infection with multiple Wolbachia STs, next-generation sequencing with improved analysis would assist with scaling up approaches to Wolbachia MLST.

Original languageEnglish
Pages (from-to)1410-1416
Number of pages7
JournalJournal of Microbiology and Biotechnology
Volume25
Issue number9
DOIs
Publication statusPublished - 2015 Jul 2

Keywords

  • Eurema
  • MLST
  • Pyrosequencing
  • Tongeia
  • Wolbachia
  • Wsp

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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