Drosophila male genital structures often wound females during mating (Kamimura 2010), and variation in wounding exists among D. sechellia w females that mate with D. mauritiana‐D. sechellia introgression males (Masly and Kamiura 2014). Thus, it is possible that female reproductive output may be affected by wounds that occur during mating. Because not all females suffer wounds during mating (Kamimura 2010; Masly and Kamimura 2014), and the presence or absence of wounds can only be determined by dissecting a female within a few days postmating, we generated wounds at the PL insertion site manually using an insect pin in both virgin and mated females. First, we collected newly enclosed D. sechellia w females and anesthetized them under light CO2, then gently inserted an unsterilized 0.25 mm diameter insect pin (Bioquip Products) between abdominal segments VII and VIII on either side of the abdomen at the site of PL insertion during copulation. An insect pin of this size substantially exceeds the size of the D. sechellia w PL, and wounds were evident by trace amounts of hemolymph that leaked out at the insertion sites. These virgin females were allowed to recover for four days in isolation before being placed in individual food vials and transferred to a new vial every three days for nine days. Control four‐day old virgin D. sechellia w females that were not wounded were likewise placed in food vials and transferred. We recorded the total number of eggs laid across all three vials. Although virgin Drosophila females do lay eggs, the presence of male Sfps in the female reproductive tract stimulates oviposition and substantially increases oviposition amounts (Wolfner 1997). Also, because D. sechellia females lay fewer eggs compared to females among D. sechellia's sister species (Coyne et al. 1991), it is possible that differences in oviposition amounts might be difficult to detect between wounded and unwounded virgin females. Therefore, we also tested the effects of wounds on oviposition in mated females. Virgin males and females were collected and aged in isolation for three days. After this time, one virgin male and one virgin female were paired together in a food vial, and the males were removed after 24 hours. Females were then lightly anesthetized using light CO2 and wounded with an insect pin as described above. Wounded experimental mated females and unwounded control mated females were returned to individual food vials and transferred to a new food vial every three days for nine days.

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