Dispatched flies were mounted and views of the ovipositor (180x) were acquired as focal stacks on an AXIO Zoom V.16 (ZEISS, Germany, Oberkochen) with a 0.5x PlanApo Z objective (ZEISS, Germany, Oberkochen). The resulting stacks were compiled to extended focus images in Helicon Focus 6 (Helicon Soft, Dominica) using the pyramid method. Here we provided images of the serrated ovipositors of close relatives to D. suzukii in order to highlight the physical deviations in egg-laying potential (Figure S1 ), which has been described previously (Atallah et al., 2014 ). We also documented the differences in male wing pigmentation (32x), as D. suzukii and D. subpulchrella can be difficult to distinguish (Figure S7 ). Moreover, D. subpulchrella were shown recently in the most up to date phylogenetic analyses of this Drosophila clade to be the closest relatives to D. suzukii, as opposed to D. biarmipes (Dekker et al., 2015 ; Hickner et al., 2016 ; Keesey et al., 2019a (link); Ramasamy et al., 2016 (link)). Images were also compiled of dissected heads (128x) to illustrate the mounting preparations for sensillum counts (Figures 2 A–2D).
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