Tandem repeat markers as novel diagnostic tools for high resolution fingerprinting of Wolbachia

Riegler, Markus, Iturbe-Ormaetxe, Inaki, Woolfit, Megan, Miller, Wolfgang J. and O'Neill, Scott L. (2012) Tandem repeat markers as novel diagnostic tools for high resolution fingerprinting of Wolbachia. BMC Microbiology, 12 s1: . doi:10.1186/1471-2180-12-S1-S12


Author Riegler, Markus
Iturbe-Ormaetxe, Inaki
Woolfit, Megan
Miller, Wolfgang J.
O'Neill, Scott L.
Title Tandem repeat markers as novel diagnostic tools for high resolution fingerprinting of Wolbachia
Formatted title
Tandem repeat markers as novel diagnostic tools for high resolution fingerprinting of Wolbachia
Journal name BMC Microbiology   Check publisher's open access policy
ISSN 1471-2180
Publication date 2012-01
Sub-type Article (original research)
DOI 10.1186/1471-2180-12-S1-S12
Open Access Status DOI
Volume 12
Issue s1
Total pages 15
Place of publication London, United Kingdom
Publisher BioMed Central
Collection year 2013
Language eng
Formatted abstract
Background
Strains of the endosymbiotic bacterium Wolbachia pipientis are extremely diverse both genotypically and in terms of their induced phenotypes in invertebrate hosts. Despite extensive molecular characterisation of Wolbachia diversity, little is known about the actual genomic diversity within or between closely related strains that group tightly on the basis of existing gene marker systems, including Multiple Locus Sequence Typing (MLST). There is an urgent need for higher resolution fingerprinting markers of Wolbachia for studies of population genetics, horizontal transmission and experimental evolution.

Results

The genome of the wMel Wolbachia strain that infects Drosophila melanogaster contains inter- and intragenic tandem repeats that may evolve through expansion or contraction. We identified hypervariable regions in wMel, including intergenic Variable Number Tandem Repeats (VNTRs), and genes encoding ankyrin (ANK) repeat domains. We amplified these markers from 14 related Wolbachia strains belonging to supergroup A and were successful in differentiating size polymorphic alleles. Because of their tandemly repeated structure and length polymorphism, the markers can be used in a PCR-diagnostic multilocus typing approach, analogous to the Multiple Locus VNTR Analysis (MLVA) established for many other bacteria and organisms. The isolated markers are highly specific for supergroup A and not informative for other supergroups. However, in silico analysis of completed genomes from other supergroups revealed the presence of tandem repeats that are variable and could therefore be useful for typing target strains.

Conclusions

Wolbachia genomes contain inter- and intragenic tandem repeats that evolve through expansion or contraction. A selection of polymorphic tandem repeats is a novel and useful PCR diagnostic extension to the existing MLST typing system of Wolbachia, as it allows rapid and inexpensive high-throughput fingerprinting of closely related strains for which polymorphic markers were previously lacking.
Keyword Cytoplasmic incompatibility expression
Rhagoletis-cerasi diptera
Sequence typing system
Locus variable-number
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
School of Biological Sciences Publications
 
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