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An efficient molecular technique for sexing tiger pufferfish (fugu) and the occurrence of sex reversal in a hatchery population

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Abstract

The tiger pufferfish (fugu) is one of the most important food fishes in East Asia. Since its testes are regarded as a delicacy, sex determination is economically relevant. Previous studies have identified a missense single-nucleotide polymorphism (SNP) in the Amhr2 (anti-Müllerian hormone receptor type II) gene as a strong candidate for a master sex-determining polymorphism. To distinguish genotypic sex efficiently, we developed a high-resolution melting (HRM) assay for this SNP site. By screening 396 fish from two independent crosses reared under controlled conditions, we observed perfect concordance between the SNP genotype and phenotypic sex. Thus, this method holds great potential for use in high-throughput sexing. When analyzing 293 progeny from a third cross reared under unknown conditions, we unexpectedly found that 25 % of phenotypic males exhibited female genotype. These results suggest that environmental factors such as rearing conditions could influence the sex-determination pathway in pufferfish. Alternatively, genetic modifiers might override the signals from Amhr2. This finding raises a concern regarding enhanced stock management of this species, because sex-reversed fish could compromise the sex ratio in subsequent generations. The HRM assay will also be useful for monitoring the degree of sex reversal before release.

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Acknowledgments

We thank Naoki Mizuno, Yuka Jo, and Masashi Fujita for technical support, and Yasutoshi Yoshiura for communicating unpublished technical data. This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture of Japan, and by grants from Research and Development Projects for Application in Promoting New Policy of Agriculture, Forestry, and Fisheries, and the Program for Promotion of Basic Research Activities for Innovative Biosciences.

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Authors and Affiliations

  1. Fisheries Laboratory, University of Tokyo, Hamamatsu, Shizuoka, 431-0214, Japan

    Takayoshi Matsunaga, Risa Ieda, Sho Hosoya, Miwa Kuroyanagi, Satoshi Tasumi, Yuzuru Suzuki & Kiyoshi Kikuchi

  2. Department of Marine Bioscience, Fukui Prefectural University, Obama, Fukui, 917-0003, Japan

    Hiroaki Suetake & Toshiaki Miyadai

  3. National Research Institute of Aquaculture, Fisheries Research Agency, Minamiizu, Shizuoka, 236-8648, Japan

    Shigenori Suzuki

Authors
  1. Takayoshi Matsunaga
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  2. Risa Ieda
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  3. Sho Hosoya
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  4. Miwa Kuroyanagi
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  5. Shigenori Suzuki
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  6. Hiroaki Suetake
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  7. Satoshi Tasumi
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  8. Yuzuru Suzuki
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  9. Toshiaki Miyadai
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  10. Kiyoshi Kikuchi
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Correspondence to Kiyoshi Kikuchi.

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T. Matsunaga and R. Ieda contributed equally to this work.

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Matsunaga, T., Ieda, R., Hosoya, S. et al. An efficient molecular technique for sexing tiger pufferfish (fugu) and the occurrence of sex reversal in a hatchery population. Fish Sci 80, 933–942 (2014). https://doi.org/10.1007/s12562-014-0768-0

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