Crossing scheme for the generation of the isochromosomal lines.

<p>One highly resistant virgin female of each original Asian fly stock was mated with two males of the multi-balancer stock. F1 generation males that carried an Ama chromosome 2 balanced over CyO and an Ama chromosome 3 balanced over TM6B, <i>Tb</i> or MKRS were crossed back to one multi-balancer virgin female. F2 generation males carrying an Ama chromosome 2 balanced over CyO and an Ama chromosome 3 balanced over TM6B, <i>Tb</i> were back-crossed to one multi-balancer virgin female. Virgin siblings of the F3 generation were then crossed to produce the isochromosomal lines.</p

Comparison between current and historic resistance values.

<p>Our calculated LC₅₀ values and how they compare to the values calculated in 1982 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0127569#pone.0127569.ref022" target="_blank">22</a>] are shown. Oregon-R served as the normalization control for the relative resistance values between today and 1982. LC₅₀ values are given in [μg of α-amanitin per g of larval food]. All values are averages of three experimental replicates.</p><p>Comparison between current and historic resistance values.</p

Adult thorax lengths of the three original Asian stocks.

<p>A) Ama-KTT, B) Ama-MI, and C) Ama-KLM. Male and female thorax lengths (y-axis) from flies that hatched from different α-amanitin concentrations (x-axis) were measured. The data resulting from three experimental replicates were pooled. The error bars represent the s.e.m.</p

Adult hatch time delay of the three original Asian stocks.

<p>A) Ama-KTT, B) Ama-MI, and C) Ama-KLM. The first-instar larvae were laid on day 0. The data resulting from three experimental replicates were pooled. The error bars represent the s.e.m.</p

LC₅₀ analyses for all fly stocks.

<p>A) Oregon-R, Canton-S, and multi-balancer stock; B) Ama-KTT, Ama-KTT/M/2, and Ama-KTT/M/5; C) Ama-MI, Ama-MI/M/2, and Ama-MI/T/6; D) Ama-KLM, Ama-KLM/M/5, and Ama-KLM/M/7 LC_<b>50</b> analyses are shown. All analyses contain three experimental replicates (100 larvae in each experiment for each concentration) and were normalized, using 0-toxin concentration as a control. The error bars represent the standard error of the mean (s.e.m.).</p

Candidate genes resulting from the 180-line GWAS on 2.0 μg/g and the 37-line GWAS.

<p>Candidate genes resulting from the 180-line GWAS on 2.0 μg/g and the 37-line GWAS.</p

Manhattan plots for the three GWAS.

<p>A) 37-line GWAS using LC₅₀ values, B) 180-line GWAS on 2.0 μg/g α-amanitin, C) 180-line GWAS on 0.2 μg/g α-amanitin. Selected significant gene names are printed on the top right of the corresponding dots in the graphs.</p

Larval viability variation in the DGRP lines in response to α-amanitin.

<p>The y-axis shows individual viability values, while the x-axis represents the individual DGRP lines. The lines are sorted from lowest α-amanitin resistance (left) to highest α-amanitin resistance (right). The error bars represent the standard error of the mean (SEM). A) 180 lines tested on 0.2 μg/g α-amanitin. (Individual line numbers are not shown but can be found in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0173162#pone.0173162.s001" target="_blank">S1 Table</a>). The y-axis represents the average number of flies hatched from 10 larvae placed on toxic food. B) 180 lines tested on 2.0 μg/g α-amanitin. (Individual line numbers are not shown but can be can be found in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0173162#pone.0173162.s001" target="_blank">S1 Table</a>). The y-axis represents the average hatch counts out of 10 larvae placed on toxic food. C). The y-axis represents the LC₅₀ values of the 37-line subset. The line numbers are shown on the x-axis.</p