Fisheries Science Identification of Male Sex-Linked DNA Sequence of the Cultured Pacific Bluefin Tuna Thunnus orientalis Yasuo Agawa a*, Mayui Iwaki a, Takafumi Komiya a, Tomoki Honryo a, Kouhei Tamura a, Tsukasa Okada b, Naoki Yagishita c, Toru Kobayashi c and Yoshifumi Sawada a a Oshima Branch, Fisheries Laboratories, Kinki University Ohshima Kushimoto, Wakayama , Japan b Hokkaido System Science Co., Ltd Shinkawa-nishi 2 jyo, Kitaku, Sapporo, Hokkaido , Japan c Department of Fisheries, Faculty of Agriculture, Kinki University, Nakamachi, Nara, Nara , Japan *To whom correspondence should be addressed. Tel Fax E- mail:

Figure S1. Agawa et al. ♂ ♀ Fisheries Science, Identification of Male Sex-Linked DNA Sequence of the Cultured Pacific Bluefin Tuna Thunnus orientalis. ○ Y. Agawa, M. Iwaki, T. Komiya, T. Honryo, K. Tamura, T. Okada, N. Yagishita, T. Kobayashi and Y. Sawada ○ To whom correspondence. Address; Fisheries Oshima Branch, Fisheries Laboratories, Kinki University Oshima, Kushimoto, Wakayama, , Japan tel

Figure S1. Agawa et al. (Countinued) ♂ ♀

♂ ♀ 6bp deletion

Figure S1. Agawa et al. (Countinued) ♂ ♀

♂ ♀

♂ ♀

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Figure S1. Agawa et al. (Countinued) Nucleotide sequence alignment of the top 30 frequent sequences obtained by high-throughput sequencing; top panel, male and bottom, female. Identical nucleotides are surrounded by a black square, nucleotide deletions are shown by (-), and substitutions are indicated by small squares and specific nucleotides. For each sequence used for the alignment, the name, read count and nucleotide length are provided to the left side of the sequence. Six bp continuous deletion was highlighted by red square around nucleotide position 170 bp. ♂ ♀

Figure S2. Agawa et al.

Figure S2. Agawa et al. (continued) Nucleotide sequence alignment of the aquacultured F2 and F3 PBF mitochondrial patial d-loop seqences. Identical nucleotides are surrounded by a black square, nucleotide deletions are shown by (-), and substitutions are indicated by small squares and specific nucleotides.

Table S1. Summary of the sequence read counting of top 30 frequent sequences in a male and a female Male Female 6 bp 6 bp Rank deletion reads length (bp) deletion reads length (bp) ○ ○ ○ ○ Total sequence reads base deleted- sequence reads base deleted / total (%) Fisheries Science, Identification of Male Sex-Linked DNA Sequence of the Cultured Pacific Bluefin Tuna Thunnus orientalis. ○ Y. Agawa, M. Iwaki, T. Komiya, T. Honryo, K. Tamura, T. Okada, N. Yagishita, T. Kobayashi and Y. Sawada ○ To whom correspondence. Address; Fisheries Oshima Branch, Fisheries Laboratories, Kinki University Oshima, Kushimoto, Wakayama, , Japan tel