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http://dx.doi.org/10.4014/jmb.1503.03097

Wolbachia Sequence Typing in Butterflies Using Pyrosequencing  

Choi, Sungmi (BK21PLUS Program in Embodiment: Health-Society Interaction, Department of Public Health Sciences, Graduate School, Korea University)
Shin, Su-Kyoung (BK21PLUS Program in Embodiment: Health-Society Interaction, Department of Public Health Sciences, Graduate School, Korea University)
Jeong, Gilsang (Department of Ecology and Evolution, National Institute of Ecology)
Yi, Hana (BK21PLUS Program in Embodiment: Health-Society Interaction, Department of Public Health Sciences, Graduate School)
Publication Information
Journal of Microbiology and Biotechnology / v.25, no.9, 2015 , pp. 1410-1416 More about this Journal
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.
Keywords
Wolbachia; Eurema; Tongeia; MLST; wsp; pyrosequencing;
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1 Werren JH, Baldo L, Clark ME. 2008. Wolbachia: master manipulators of invertebrate biology. Nat. Rev. Microbiol. 6: 741-751.   DOI
2 Zhou W, Rousset F, O'Neil S. 1998. Phylogeny and PCRbased classification of Wolbachia strains using wsp gene sequences. Proc. Biol. Sci. 265: 509-515.   DOI
3 Salunke BK, Salunkhe RC, Dhotre DP, Khandagale AB, Walujkar SA, Kirwale GS, et al. 2010. Diversity of Wolbachia in Odontotermes spp. (Termitidae) and Coptotermes heimi (Rhinotermitidae) using the multigene approach. FEMS Microbiol. Lett. 307: 55-64.   DOI
4 Salunke BK, Salunkhe RC, Dhotre DP, Walujkar SA, Khandagale AB, Chaudhari R, et al. 2012. Determination of Wolbachia diversity in butterflies from Western Ghats, India, by a multigene approach. Appl. Environ. Microbiol. 78: 4458-4467.   DOI
5 Salunkhe RC, Narkhede KP, Shouche YS. 2014. Distribution and evolutionary impact of Wolbachia on butterfly hosts. Ind. J. Microbiol. 54: 249-254.   DOI
6 Lee SH, Kwon SG, Nam GP, Son JD, Lee JG, Park YK, et al. 2012. Studies on ecological environments and indoor-rearing conditions of the common grass yellow butterfly, Eurema hecabe. J. Seric. Entomol. Sci. 50: 133-139.
7 Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB, et al. 2009. Introducing Mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl. Environ. Microbiol. 75: 7537-7541.   DOI
8 Tagami Y, Miura K. 2004. Distribution and prevalence of Wolbachia in Japanese populations of Lepidoptera. Insect Mol. Biol. 13: 359-364.   DOI
9 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. Mol. Biol. Evol. 28: 2731-2739.   DOI
10 Weeks AR, Breeuwer JA. 2001. Wolbachia-induced parthenogenesis in a genus of phytophagous mites. Proc. Biol. Sci. 268: 2245-2251.   DOI
11 Narita S, Kageyama D, Nomura M, Fukatsu T. 2007. 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: 4332-4341.   DOI
12 Narita S, Nomura M, Kageyama D. 2007. 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: 235-245.   DOI
13 Narita S, Nomura M, Kato Y, Yata O, Kageyama D. 2007. Molecular phylogeography of two sibling species of Eurema butterflies. Genetica 131: 241-253.   DOI
14 Riegler M, Stauffer C. 2002. Wolbachia infections and superinfections in cytoplasmically incompatible populations of the European cherry fruit fly Rhagoletis cerasi (Diptera, Tephritidae). Mol. Ecol. 11: 2425-2434.   DOI
15 O'Neill SL, Giordano R, Colbert AM, Karr TL, Robertson HM. 1992. 16S rRNA phylogenetic analysis of the bacterial endosymbionts associated with cytoplasmic incompatibility in insects. Proc. Natl. Acad. Sci. USA 89: 2699-2702.   DOI
16 Perez-Losada M, Cabezas P, Castro-Nallar E, Crandall KA. 2013. Pathogen typing in the genomics era: MLST and the future of molecular epidemiology. Infect. Genet. Evol. 16: 38-53.   DOI
17 Raychoudhury R, Baldo L, Oliveira DC, Werren JH. 2009. Modes of acquisition of Wolbachia: horizontal transfer, hybrid introgression, and codivergence in the Nasonia species complex. Evolution 63: 165-183.   DOI
18 Ros VI, Fleming VM, Feil EJ, Breeuwer JA. 2009. 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: 1036-1043.   DOI
19 Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R. 2011. UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 27: 2194-2200.   DOI
20 Gilles A, Meglecz E, Pech N, Ferreira S, Malausa T, Martin JF. 2011. Accuracy and quality assessment of 454 GS-FLX titanium pyrosequencing. BMC Genomics 12: 245.   DOI
21 Goodacre SL, Martin OY, Thomas CF, Hewitt GM. 2006. Wolbachia and other endosymbiont infections in spiders. Mol. Ecol. 15: 517-527.   DOI
22 Ilinsky Y. 2012. Full information on strain Tfis_B. Available at http://pubmlst.org/perl/bigsdb/bigsdb.pl?db=pubmlstwolbachia isolates&page=info&id=474.
23 Hiroki M, Kato Y, Kamito T, Miura K. 2002. Feminization of genetic males by a symbiotic bacterium in a butterfly, Eurema hecabe (Lepidoptera: Pieridae). Naturwissenschaften 89: 167-170.   DOI
24 Huelsenbeck JP, Ronquist F. 2001. MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics 17: 754-755.   DOI
25 Huse SM, Huber JA, Morrison HG, Sogin ML, Welch DM. 2007. Accuracy and quality of massively parallel DNA pyrosequencing. Genome Biol. 8: R143.   DOI
26 Jeratthitikul E, Hara T, Yago M, Itoh T, Wang M, Usami S, Hikida T. 2013. Phylogeography of Fischer’s blue, Tongeia fischeri, in Japan: evidence for introgressive hybridization. Mol. Phylogenet. Evol. 66: 316-326.   DOI
27 Kikuchi Y, Fukatsu T. 2003. Diversity of Wolbachia endosymbionts in heteropteran bugs. Appl. Environ. Microbiol. 69: 6082-6090.   DOI
28 Kunin V, Engelbrektson A, Ochman H, Hugenholtz P. 2010. Wrinkles in the rare biosphere: pyrosequencing errors can lead to artificial inflation of diversity estimates. Environ. Microbiol. 12: 118-123.   DOI
29 Atyame CM, Delsuc F, Pasteur N, Weill M, Duron O. 2011. Diversification of Wolbachia endosymbiont in the Culex pipiens mosquito. Mol. Biol. Evol. 28: 2761-2772.   DOI
30 Baldo L, Dunning Hotopp JC, Jolley KA, Bordenstein SR, Biber SA, Choudhury RR, et al. 2006. Multilocus sequence typing system for the endosymbiont Wolbachia pipientis. Appl. Environ. Microbiol. 72: 7098-7110.   DOI
31 carol lb, ward bw, ehrlich pr. 2003. butterlies: ecology and Evolution Taking Flight. University of Chicago Press, IL.
32 Bing XL, Xia WQ, Gui JD, Yan GH, Wang XW, Liu SS. 2014. Diversity and evolution of the Wolbachia endosymbionts of Bemisia (Hemiptera: Aleyrodidae) whiteflies. Ecol. Evol. 4: 2714-2737.   DOI