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These include families associated with carboxylesterase activity (GO:0004091) and guanyl-nucleotide exchange factor activity (GO:0005085). Thus, these gene families have accumulated nsSNPs and cInDels in the Dark-fly genome and might be potentially related to the Dark-fly’s traits. 01) were listed for Dark-fly using the DAVID tool. * indicates a family shared with that listed for Oregon-R-S (data not shown). 3 Nonsense Mutations in the Dark-fly Genome Among nsSNPs, a nonsense mutation produces a stop codon in the amino acid sequence of a gene product, and may severely affect the protein’s function (Fig.

5 % of the SNPs were located within protein coding regions of genes. It is not easy to evaluate SNPs in non-coding regions, and accordingly we focused on the coding SNPs. , they do not alter amino acid sequences of gene products) (Fig. , they change the amino acid sequence) (Fig. 5c). We collected the Dark-fly-specific nsSNPs without redundancy between isoforms and identified 4,323 genes carrying nsSNPs. 2). We performed similar processes for the Oregon-R-S genome and identified 3,039 such genes.

Dark-fly males and females copulated more quickly than the Oregon-R-S pairs (Fig. 6 min for Oregon-R-S), suggesting that mating behaviors might be stimulated in the Dark-fly pairs: males might be more active for courtship and/or females might more easily accept males. Since the quick copulation of Dark-fly was observed in light conditions as well as in dark conditions (Fig. 3), the quick copulation alone would not account for the early reproduction in the dark. However, we speculate that stimulated sexual behavior contributes to the early reproduction via re-courtship after failure and/or via repeated mating.

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