1. 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 1790-4 Ohshima Kushimoto, Wakayama 649-3633, Japan b Hokkaido System Science Co., Ltd. 1-2-1 Shinkawa-nishi 2 jyo, Kitaku, Sapporo, Hokkaido 001- 0932, Japan c Department of Fisheries, Faculty of Agriculture, Kinki University, 3327-204 Nakamachi, Nara, Nara 631-8505, Japan *To whom correspondence should be addressed. Tel. +81-735-65-0501. Fax. +81 -735-65-0974. E- mail: agawa@kindaisuiken.jpagawa@kindaisuiken.jp

2. 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 1790-4 Oshima, Kushimoto, Wakayama, 649-3633, Japan tel +81-735-65-0501

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

4. ♂ ♀ 6bp deletion

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

6. ♂ ♀

7. ♂ ♀

8. ♂ ♀

9. 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. ♂ ♀

10. Figure S2. Agawa et al.

11. 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.

12. 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) 1 983 413 930 415 2 548 416 707 415 3 477 415 640 414 4 425 414 337 416 5 ○ 419 408 221 415 6 374 416 183 417 7 345 415 144 415 8 315 415 143 416 9 291 417 138 414 10 281 413 123 411 11 278 415 118 415 12 270 415 111 412 13 ○ 238 405 106 412 14 ○ 213 407 96 413 15 203 414 88 415 16 194 414 88 414 17 188 414 86 415 18 181 414 83 415 19 178 415 82 415 20 175 414 82 418 21 154 412 79 415 22 153 415 79 413 23 ○ 153 405 79 413 24 146 415 79 415 25 137 413 78 418 26 131 415 76 414 27 123 415 73 415 28 122 413 71 413 29 119 413 68 413 30 118 411 68 410 Total sequence reads 7932 5256 6 base deleted- sequence reads 1023 0 6 base deleted / total (%) 12.8 0 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 1790-4 Oshima, Kushimoto, Wakayama, 649-3633, Japan tel +81-735-65-0501