Wolbachia Variants Induce Differential Protection to Viruses in Drosophila melanogaster: A Phenotypic and Phylogenomic Analysis
Figure 4
Wolbachia densities and Drosophila longevity are variant dependent.
(A) 3–4 and 6–7 day old males of each wMel variant line and w1118 iso were collected for DNA extraction and qPCR. Relative amount of Wolbachia genomic DNA was calculated using host Rpl32 as a reference gene and values are relative to median of 3–4 days-old wMelCS_b samples. Each point represents a replicate (ten males per replicate, four replicates per Drosophila line), and lines are medians of the replicates. The compact letters display of pairwise Wilcoxon rank sum tests between variants is shown on the top. (B) The survival of one hundred males of each wMel variant line and w1118 iso was checked every five days. One more replicate was performed with similar results. (C) Cox hazard ratio of each wMel variant line compared to w1118 iso. The natural logarithm of the Cox hazard ratio is shown. Error bars represent standard error. Letters refer to compact letter display of Tukey's test of all pairwise comparisons. Analysis is based on two independent replicates, each with 100 flies per line, with 10 flies per vial. w1118 iso is assigned to group “a” in the compact letter display of Tukey's test (not shown). (D) Males of wMel, wMelCS_a and wMelCS_b lines were collected for DNA extraction and qPCR every 10 days. Day 0 corresponds to 3–6 days-old flies, wMelCS_a were collected up to 40 days and wMel and wMelCS_b up to 50 days. There are no further time points due to high mortality. Each point represents a replicate (ten males per replicate, five replicates per time point), and lines are medians of the replicates. Relative amount of Wolbachia genomic DNA was calculated using host Rpl32 as a reference gene and values are relative to median of samples of wMelCS_b at day zero. (E) The survival of one hundred males of each tetracycline treated line derived from the wMel variants lines and w1118 iso was checked every five days. The experiment was repeated once with similar results. (F) Cox hazard ratio of each tetracycline treated line, derived from the wMel variants lines, compared to w1118 iso tetracycline treated line. Analysis is based on two independent replicates, each with 100 flies per line, with 10 flies per vial. w1118 iso tetracycline treated line is assigned to group “d” in the compact letter display of Tukey's test (not shown).