Detection of genetic sex using a qPCR-based method in the bigfin reef squid (Sepioteuthis lessoniana)
The bigfin reef squid (Sepioteuthis lessoniana) is a valuable commercial species, especially in East Asia. While small-scale cultivation exists, efforts to achieve large-scale aquaculture are underway. However, full-life cycle aquaculture remains challenging due to reproductive issues, particularly...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | Aquaculture Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352513425000304 |
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| author | Yi-Chien Lee Li-Lun Tang Yu-Chen Yeh Hau-Wen Li Huei-Mien Ke Cheng-Ju Yang Chun-I Chiang Chia-Hui Wang Yung-Che Tseng Ching-Fong Chang Isheng Jason Tsai Guan-Chung Wu |
| author_facet | Yi-Chien Lee Li-Lun Tang Yu-Chen Yeh Hau-Wen Li Huei-Mien Ke Cheng-Ju Yang Chun-I Chiang Chia-Hui Wang Yung-Che Tseng Ching-Fong Chang Isheng Jason Tsai Guan-Chung Wu |
| author_sort | Yi-Chien Lee |
| collection | DOAJ |
| description | The bigfin reef squid (Sepioteuthis lessoniana) is a valuable commercial species, especially in East Asia. While small-scale cultivation exists, efforts to achieve large-scale aquaculture are underway. However, full-life cycle aquaculture remains challenging due to reproductive issues, particularly in later generations, where abnormal offspring production is common. One possible issue is sex reversal under culture conditions, possibly due to high temperatures and smaller egg sizes in later generations. Therefore, a reliable method to determine the genetic sex of bigfin reef squid is crucial. Analysis of the female and male genome coverage along the assembly confirms a ZZ/Z0 female hemizygous sex-determination system in bigfin reef squid. We identified Z chromosome linked regions and developed a quantitative PCR (qPCR)-based method to measure the copy number variance of these Z-specific regions, normalized by autosomal regions, with ratios of 1:0.5 in ZZ males and Z0 females, respectively. Among the primer pairs designed for qPCR, two primer pairs in Z-specific regions and one primer pair in autosomal region exhibited a single melting curve (indicating the predicted same fragment) across all lineages. This genetic sexing method was validated in samples from different populations across Taiwan, demonstrating its effectiveness in determining the genetic sex of bigfin reef squid. Our findings provide a valuable tool for aquaculture management and offer new insights into sex determination mechanisms in cephalopods. |
| format | Article |
| id | doaj-art-accc7ca354dd4a9684893d0d773f224d |
| institution | Kabale University |
| issn | 2352-5134 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Aquaculture Reports |
| spelling | doaj-art-accc7ca354dd4a9684893d0d773f224d2025-02-06T05:12:25ZengElsevierAquaculture Reports2352-51342025-03-0140102644Detection of genetic sex using a qPCR-based method in the bigfin reef squid (Sepioteuthis lessoniana)Yi-Chien Lee0Li-Lun Tang1Yu-Chen Yeh2Hau-Wen Li3Huei-Mien Ke4Cheng-Ju Yang5Chun-I Chiang6Chia-Hui Wang7Yung-Che Tseng8Ching-Fong Chang9Isheng Jason Tsai10Guan-Chung Wu11Biodiversity Research Center, Academia Sinica, Taipei 115, TaiwanDepartment of Aquaculture, National Taiwan Ocean University, Keelung 202, TaiwanBiodiversity Research Center, Academia Sinica, Taipei 115, Taiwan; Genome and Systems Biology Degree Program, Academia Sinica and National Taiwan University, Taipei 106, TaiwanDepartment of Aquaculture, National Taiwan Ocean University, Keelung 202, TaiwanDepartment of Microbiology, Soochow University, Taipei 111, TaiwanBiodiversity Research Center, Academia Sinica, Taipei 115, TaiwanDepatment of Environmental Biology and Fisheries Science, National Taiwan Ocean University, Keelung 202, TaiwanDepatment of Environmental Biology and Fisheries Science, National Taiwan Ocean University, Keelung 202, TaiwanMarine Research Station, Institute of Cellular and Organismic Biology, Academia Sinica, Yilan 262, TaiwanCenter of Excellence for the Oceans, National Taiwan Ocean University, Keelung 202, TaiwanBiodiversity Research Center, Academia Sinica, Taipei 115, Taiwan; Corresponding author.Department of Aquaculture, National Taiwan Ocean University, Keelung 202, Taiwan; Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung 202, Taiwan; Corresponding author at: Department of Aquaculture, National Taiwan Ocean University, Keelung 202, Taiwan.The bigfin reef squid (Sepioteuthis lessoniana) is a valuable commercial species, especially in East Asia. While small-scale cultivation exists, efforts to achieve large-scale aquaculture are underway. However, full-life cycle aquaculture remains challenging due to reproductive issues, particularly in later generations, where abnormal offspring production is common. One possible issue is sex reversal under culture conditions, possibly due to high temperatures and smaller egg sizes in later generations. Therefore, a reliable method to determine the genetic sex of bigfin reef squid is crucial. Analysis of the female and male genome coverage along the assembly confirms a ZZ/Z0 female hemizygous sex-determination system in bigfin reef squid. We identified Z chromosome linked regions and developed a quantitative PCR (qPCR)-based method to measure the copy number variance of these Z-specific regions, normalized by autosomal regions, with ratios of 1:0.5 in ZZ males and Z0 females, respectively. Among the primer pairs designed for qPCR, two primer pairs in Z-specific regions and one primer pair in autosomal region exhibited a single melting curve (indicating the predicted same fragment) across all lineages. This genetic sexing method was validated in samples from different populations across Taiwan, demonstrating its effectiveness in determining the genetic sex of bigfin reef squid. Our findings provide a valuable tool for aquaculture management and offer new insights into sex determination mechanisms in cephalopods.http://www.sciencedirect.com/science/article/pii/S2352513425000304Sex determinationCephalopodSex-linked markerHemizygous sex chromosome |
| spellingShingle | Yi-Chien Lee Li-Lun Tang Yu-Chen Yeh Hau-Wen Li Huei-Mien Ke Cheng-Ju Yang Chun-I Chiang Chia-Hui Wang Yung-Che Tseng Ching-Fong Chang Isheng Jason Tsai Guan-Chung Wu Detection of genetic sex using a qPCR-based method in the bigfin reef squid (Sepioteuthis lessoniana) Aquaculture Reports Sex determination Cephalopod Sex-linked marker Hemizygous sex chromosome |
| title | Detection of genetic sex using a qPCR-based method in the bigfin reef squid (Sepioteuthis lessoniana) |
| title_full | Detection of genetic sex using a qPCR-based method in the bigfin reef squid (Sepioteuthis lessoniana) |
| title_fullStr | Detection of genetic sex using a qPCR-based method in the bigfin reef squid (Sepioteuthis lessoniana) |
| title_full_unstemmed | Detection of genetic sex using a qPCR-based method in the bigfin reef squid (Sepioteuthis lessoniana) |
| title_short | Detection of genetic sex using a qPCR-based method in the bigfin reef squid (Sepioteuthis lessoniana) |
| title_sort | detection of genetic sex using a qpcr based method in the bigfin reef squid sepioteuthis lessoniana |
| topic | Sex determination Cephalopod Sex-linked marker Hemizygous sex chromosome |
| url | http://www.sciencedirect.com/science/article/pii/S2352513425000304 |
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