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Identification of Male-Specific Molecular Marker and Development of PCR-Based Genetic Sex Identification Technique in Spotted Knifejaw (Oplegnathus punctatus)

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Abstract

Spotted knifejaw (Oplegnathus punctatus) is a marine teleost species that is economically important for aquaculture and marine pasture proliferation and shows obvious bisexual growth dimorphism, but molecular sex markers are currently lacking. A 290 bp (base pair) insertion with two fragments (230 bp and 60 bp) was identified in male individuals of O. punctatus based on whole-genome sequencing scanning and structural variation analyses. The gene annotation results showed that the insertion event occurred in the Igfn1 gene of male O. punctatus. The results of amino acid analysis further showed that the insertion event resulted in the functional variation of Igfn1 in male O. punctatus, and recombination caused the inactivation of Igfn1. According to the male-specific insertion information, we designed a PCR-based genetic amplification technique for rapid sex identification in O. punctatus. The results of agarose gel electrophoresis showed that two DNA fragments of 635 bp and 925 bp were amplified in male O. punctatus, while only a single DNA fragment of 635 bp was amplified in female individuals. The sex of individuals identified by this method was consistent with their known phenotypic sex, which will improve sex identification efficiency. This method provides a new DNA marker for rapid sex identification in O. punctatus, which has great significance and application value in monosex breeding and provides new insights for the study of Igfn1 gene recombination and inactivation in male O. punctatus.

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Acknowledgements

The authors thank the Institute of Oceanology, the Chinese Academy of Sciences for the infrastructural support, Prof. Zhizhong Xiao for the experimental materials, and Prof. Jun Li for his guidance.

Funding

This work was supported by grants from the National Key Research and Development Program (2018YFD0901204), Key Special Project for Introduced Talents Team of Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou) (GML2019ZD0402), Key Deployment Projects of Center for Ocean Mega-Science, Chinese Academy of Sciences (Frontier Cross-category, COMS2020Q05), China Agriculture Research System (CARS-47), Major Agricultural Application Technology Innovation Project of Shandong Province (SD2019YY01), STS project (KFZD-SW-106, ZSSD-019, 2017T3017, and KFJ-STS QYZX-020), Qingdao National Laboratory for Marine Science and Technology (2018SDKJ0502-2 and 2015ASKJ02), and National Natural Science Foundation of China (No. 31672672).

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Author notes

  1. Yuting Ma and Jun Li contributed equally.

Authors and Affiliations

  1. CAS and Shandong Province Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China

    Yuting Ma, Yongshuang Xiao, Zhizhong Xiao, Yanduo Wu, Haixia Zhao & Jun Li

  2. School of Marine Science and Engineering, Qingdao Agricultural University, Qingdao, China

    Yuting Ma

  3. Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou), Guangzhou, China

    Yongshuang Xiao, Zhizhong Xiao & Jun Li

  4. Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China

    Yongshuang Xiao, Zhizhong Xiao & Jun Li

  5. College of Marine Science, University of Chinese Academy of Sciences, Beijing, China

    Zhizhong Xiao, Yanduo Wu & Haixia Zhao

  6. Weihai Haohuigan Marine Biotechnology Co, Weihai, 264400, China

    Zhizhong Xiao

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Contributions

YSX and JL conceived and designed the project and revised the manuscript. YTM, JL and YSX performed the genomic investigations and wrote the manuscript. ZZX, YDW, and HXZ participated in data analysis, discussion, and figure preparation. All authors read and approved the final manuscript.

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Correspondence to Yongshuang Xiao.

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Ma, Y., Xiao, Y., Xiao, Z. et al. Identification of Male-Specific Molecular Marker and Development of PCR-Based Genetic Sex Identification Technique in Spotted Knifejaw (Oplegnathus punctatus). Mar Biotechnol 24, 969–978 (2022). https://doi.org/10.1007/s10126-022-10160-w

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