16S rRNA gene profiling of D

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16S rRNA gene profiling of D 16S rRNA gene profiling of D. melanogaster shows reduction of Acetobacteraceae levels in Wolbachia-infected adult flies. 16S rRNA gene profiling of D. melanogaster shows reduction of Acetobacteraceae levels in Wolbachia-infected adult flies. (A) Grape juice agar plates used for D. mauritiana egg collection. Grape juice acts as a pH indicator, turning pink under acidic conditions and yellow under basic conditions. Over time, the microbial composition of the feces of Wolbachia-infected flies turns plates more yellowish than plates used by Wolbachia-free flies. (B) Schematic of the bacterial 16S rRNA gene. Hypervariable regions V1 to V9 are red. Primers (arrows, 27F and 338R) amplify regions V1 and V2. In BstZ17I restriction enzyme digestion of the total 16S rRNA gene pool, only the Wolbachia rRNA gene between V1 and V2 is selectively digested. (C) Agarose gel image showing the 16S rRNA gene PCR products from the microbiome of D. melanogaster and efficient digestion of the Wolbachia 16S rRNA gene amplicon by the BstZ17I restriction enzyme. The red arrowhead indicates the digested Wolbachia product. The pie charts indicate the percentages of Wolbachia reads before and after BstZ17I digestion. (D) 16S rRNA gene profiles of male and female wMel-free (W−) or -infected D. melanogaster. The proportions of Acetobacteraceae are significantly different under every pair of conditions (P < 0.0001, chi-square test with Yates correction). Rama K. Simhadri et al. mSphere 2017; doi:10.1128/mSphere.00287-17