Wolbachia Sequence Typing in Butterflies Using Pyrosequencing
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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.
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.
참고문헌 (32)
- Atyame CM, "Diversification of Wolbachia endosymbiont in the Culex pipiens mosquito", Mol. Biol. Evol, 28, pp.2761~2772 (2011)
- Baldo L, "Multilocus sequence typing system for the endosymbiont Wolbachia pipientis", Appl. Environ. Microbiol, 72, pp.7098~7110 (2006)
- Bing XL, "Diversity and evolution of the Wolbachia endosymbionts of Bemisia (Hemiptera: Aleyrodidae) whiteflies", Ecol. Evol, 4, pp.2714~2737 (2014)
- carol lb, "butterlies: ecology and Evolution Taking Flight" (2003), University of Chicago Press
- Edgar RC, "UCHIME improves sensitivity and speed of chimera detection", Bioinformatics, 27, pp.2194~2200 (2011)
- Gilles A, "Accuracy and quality assessment of 454 GS-FLX titanium pyrosequencing", BMC Genomics, 12, pp.245~ (2011)
- Goodacre SL, "Wolbachia and other endosymbiont infections in spiders", Mol. Ecol, 15, pp.517~527 (2006)
- Hiroki M, "Feminization of genetic males by a symbiotic bacterium in a butterfly, Eurema hecabe (Lepidoptera: Pieridae)", Naturwissenschaften, 89, pp.167~170 (2002)
- Huelsenbeck JP, "MRBAYES: Bayesian inference of phylogenetic trees", Bioinformatics, 17, pp.754~755 (2001)
- Huse SM, "Accuracy and quality of massively parallel DNA pyrosequencing", Genome Biol, 8, pp.R143~ (2007)
- "Full information on strain Tfis_B" (2012)[URL]
- Jeratthitikul E, "Phylogeography of Fischer’s blue, Tongeia fischeri, in Japan: evidence for introgressive hybridization", Mol. Phylogenet. Evol, 66, pp.316~326 (2013)
- Kikuchi Y, "Diversity of Wolbachia endosymbionts in heteropteran bugs", Appl. Environ. Microbiol, 69, pp.6082~6090 (2003)
- Kunin V, "Wrinkles in the rare biosphere: pyrosequencing errors can lead to artificial inflation of diversity estimates", Environ. Microbiol, 12, pp.118~123 (2010)
- Narita S, "Unexpected mechanism of symbiont-induced reversal of insect sex: feminizing Wolbachia continuously acts on the butterfly Eurema hecabe during larval development", Appl. Environ. Microbiol, 73, pp.4332~4341 (2007)
- Narita S, "Naturally occurring single and double infection with Wolbachia strains in the butterfly Eurema hecabe: transmission efficiencies and population density dynamics of each Wolbachia strain", FEMS Microbiol. Ecol, 61, pp.235~245 (2007)
- Narita S, "Molecular phylogeography of two sibling species of Eurema butterflies", Genetica, 131, pp.241~253 (2007)
- O'Neill SL, "16S rRNA phylogenetic analysis of the bacterial endosymbionts associated with cytoplasmic incompatibility in insects", Proc. Natl. Acad. Sci. USA, 89, pp.2699~2702 (1992)
- Perez-Losada M, "Pathogen typing in the genomics era: MLST and the future of molecular epidemiology", Infect. Genet. Evol, 16, pp.38~53 (2013)
- Raychoudhury R, "Modes of acquisition of Wolbachia: horizontal transfer, hybrid introgression, and codivergence in the Nasonia species complex", Evolution, 63, pp.165~183 (2009)
- Riegler M, "Wolbachia infections and superinfections in cytoplasmically incompatible populations of the European cherry fruit fly Rhagoletis cerasi (Diptera, Tephritidae)", Mol. Ecol, 11, pp.2425~2434 (2002)
- Ros VI, "How diverse is the genus Wolbachia? Multiple-gene sequencing reveals a putatively new Wolbachia supergroup recovered from spider mites (Acari: Tetranychidae)", Appl. Environ. Microbiol, 75, pp.1036~1043 (2009)
- Salunke BK, "Diversity of Wolbachia in Odontotermes spp. (Termitidae) and Coptotermes heimi (Rhinotermitidae) using the multigene approach", FEMS Microbiol. Lett, 307, pp.55~64 (2010)
- Salunke BK, "Determination of Wolbachia diversity in butterflies from Western Ghats, India, by a multigene approach", Appl. Environ. Microbiol, 78, pp.4458~4467 (2012)
- Salunkhe RC, "Distribution and evolutionary impact of Wolbachia on butterfly hosts", Ind. J. Microbiol, 54, pp.249~254 (2014)
- Lee SH, "Studies on ecological environments and indoor-rearing conditions of the common grass yellow butterfly, Eurema hecabe", J. Seric. Entomol. Sci, 50, pp.133~139 (2012)
- Schloss PD, "Introducing Mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities", Appl. Environ. Microbiol, 75, pp.7537~7541 (2009)
- Tagami Y, "Distribution and prevalence of Wolbachia in Japanese populations of Lepidoptera", Insect Mol. Biol, 13, pp.359~364 (2004)
- Tamura K, "MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods", Mol. Biol. Evol, 28, pp.2731~2739 (2011)
- Weeks AR, "Wolbachia-induced parthenogenesis in a genus of phytophagous mites", Proc. Biol. Sci, 268, pp.2245~2251 (2001)
- Werren JH, "Wolbachia: master manipulators of invertebrate biology", Nat. Rev. Microbiol, 6, pp.741~751 (2008)
- Zhou W, "Phylogeny and PCRbased classification of Wolbachia strains using wsp gene sequences", Proc. Biol. Sci, 265, pp.509~515 (1998)